ANNUAL REPORT 2016 ANNUAL REPORT 2016 ANNUAL REPORT 2016 4 VICE-DIRECTION OF BASIC RESEARCH 14 CONTENTS Molecular Oncology Programme 18 Tumour Suppression Group 20 Experimental Oncology Group 24 Telomeres and Telomerase Group 28 Cell Division and Cancer Group 32 Genomic Instability Group 36 Chromosome Dynamics Group 40 DNA Replication Group 44 Melanoma Group 48 Microenvironment & Metastasis Junior Group 52 Brain Metastasis Junior Group 54 Metabolism and Cell Signalling Junior Group 56 Cancer Cell Biology Programme 58 Genes, Development and Disease Group 60 Epithelial Carcinogenesis Group 64 Epithelial Cell Biology Junior Group 68 Growth Factors, Nutrients and Cancer Junior Group 70 Seve Ballesteros Foundation-CNIO Brain Tumour Junior Group 72 Structural Biology and Biocomputing Programme 74 Structural Computational Biology Group 76 Cell Signalling and Adhesion Junior Group 80 Structural Bases of Genome Integrity Junior Group 82 Spectroscopy and Nuclear Magnetic Resonance Unit 84 Bioinformatics Unit 86 National Bioinformatics Institute Unit 88 Electron Microscopy Unit 90 Crystallography and Protein Engineering Unit 92 VICE-DIRECTION OF TRANSLATIONAL RESEARCH 94 Human Cancer Genetics Programme 98 Human Genetics Group 100 Hereditary Endocrine Cancer Group 104 Genetic and Molecular Epidemiology Group 108 Familial Cancer Clinical Unit 112 Molecular Cytogenetics and Genome Editing Unit 114 Human Genotyping-CEGEN Unit 116 Clinical Research Programme 118 Gastrointestinal Cancer Clinical Research Unit 120 Breast Cancer Junior Clinical Research Unit 122 Prostate Cancer Junior Clinical Research Unit 124 Molecular Diagnostics Unit 126 H12O-CNIO Haematological Malignancies Clinical Research Unit 128 H12O-CNIO Lung Cancer Clinical Research Unit 130 Biobank 132 FOREWORD 9 ORGANISATION OF RESEARCH 12 SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 5 DIRECTION OF INNOVATION 134 Biotechnology Programme 138 Genomics Core Unit 140 Transgenic Mice Core Unit 142 Monoclonal Antibodies Core Unit 144 Molecular Imaging Core Unit 146 Flow Cytometry Core Unit 148 Confocal Microscopy Core Unit 150 Proteomics Core Unit 152 Histopathology Core Unit 154 Animal Facility 156 Experimental Therapeutics Programme 158 Medicinal Chemistry Section 160 Biology Section 164 CNIO - Lilly Cell Signalling Therapies Section 168 CNIO - Lilly Epigenetics Section 170 Technology Transfer and Valorisation Office 172 Private Sponsors 174 COMMUNICATION 176 INTERNATIONAL AFFAIRS 186 CNIO OFFICES 190 Dean’s Office 192 CNIO Women in Science Office 194 FACTS & FIGURES 196 Scientific Management 198 Competitive Funding 200 Education and Training Programmes 214 Scientific Events 220 Administration 234 Board of Trustees 234 Scientific Advisory Board 236 Management 238 CNIO Personnel 2016 240 CNIO FRIENDS 244 CNIO Friends 247 ‘ CNIO Friends ’ Postdoctoral Contracts 248 Juegaterapia Foundation-‘ CNIO Friends ’ 249 CNIO opens its doors to ‘CNIO Friends’ 250 Benefactor Friends/Sponsor Friends 252 Donations to the CNIO 253 CREATIVE TEAM 254 ANNUAL REPORT 2016 6 SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 7 “ Commercialisation and return on investment in research remain priorities. CNIO’s royalty income in 2016 is double that of 2012. A total of 44 inventors, about 10% of the researchers at CNIO, have contributed and benefited from this achievement.” MARIA A. BLASCO Director SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 9 FOREWORD This year, once again, the data on CNIO’s scientific performance illustrate that we are doing a superb job contributing towards scientific breakthroughs. During 2016, the CNIO authored a total of 239 papers, 28 of which were published in journals with impact factors ( IF ) ranging from 10 to 15, and 32 papers in journals with impact factors greater than 15. Comparison with previous years ( 2006-2016 ) indicates that CNIO has continued to increase the numbers of papers published in top journals with IF greater than 15. With the aim of upholding the CNIO’s strategy to remain an international Research Centre of Excellence and an influential institution in cancer research worldwide, 2016 saw the establishment of a CNIO Department of International Affairs. This new Department is headed by Carolina Pola who was the Editor of Nature Medicine for over seven years, as well as the former Director of Communications at Pharmamar, the most successful Spanish pharma company devoted to anti-tumour therapies. Since the creation of the Department of International Affairs, Carolina has redefined our international approach to embed the European concept of ‘ Responsible Research and Innovation ’ ( RRI ) in our institutional and research core. This new perspective, alongside several other efforts made by the Department, have resulted in several international collaborations that will bear fruit next year in the form of joint scientific symposiums to strengthen our research groups, as well as a plan to boost our recruitment of international talent. Our capacity to attract and retain scientific talent, providing new investigators with an interdisciplinary and multicultural environment, has become an integral part of our international agenda, which we will continue to nurture in the upcoming years. I am also happy to mention that we continue to attract new and talented young group leaders to the CNIO. After the incorporation, in 2015, of two Junior Groups working in metastasis ( led by Hector Peinado and Manuel Valiente ) in the Molecular Oncology Programme, we welcomed another new Junior Group to the Programme in 2016 : the Metabolism and Cell Signalling Group, led by Alejo Efeyan who joined the CNIO at the beginning of 2016 Maria A. Blasco Director FOREWORD ANNUAL REPORT 2016 10 FOREWORD after a brilliant postdoc with David Sabatini at the Massachusetts Institute of Technology ( MIT, Boston, USA ). During 2016, we also completed the selection process to incorporate two new Groups in the Structural Biology and Biocomputing Programme, with the aim of reinforcing lines of research in structural biology at the CNIO. The two new Junior Group Leaders who will start at CNIO in 2017 are : Ivan Plaza Menacho, currently Senior Research associate at the Structural Biology Laboratory at the Biozentrum, Univesity of Basel, Switzerland ; and Rafael Fernández Leiro, from the MRC Laboratory of Molecular Biology, Cambridge, UK. In 2016, we also created a new service unit at the CNIO : the Biological Text Mining Unit, which is focused on the application and development of biomedical text mining technologies that are becoming a key tool for the efficient exploitation of information contained in unstructured data repositories. This Unit is fully funded through the ‘ Plan de Impulso de las Tecnologías del Lenguaje de la Agenda Digital ( PITL )’, in the framework of an agreement (‘ encomienda ’) between the Secretary of State of Telecommunications of the Spanish Ministry of Energy, Tourism and Digital Agenda ( MINETAD ) and the CNIO. We also revised the organisation of the two Bioinformatics Units that were providing support and expertise to our investigators ; namely, the Bioinformatics Unit of the Structural Biology and Biocomputing Programme, and the Translational Bioinformatics Unit of the Clinical Research Programme. Under a new single structure, the CNIO Bioinformatics Unit, headed by Fátima Al Shahrour, we have joined the efforts of both previous units in order to better serve the needs of the CNIO Research Groups and to better coordinate and manage bioinformatics projects. The valorisation of the research results generated by CNIO’s scientists, with the aim of turning them into high-potential diagnostic or therapeutic products and services, is one of our ways of creating added value for society and boosting public benefit through improving cancer patient outcomes, in particular. Óscar Fernández Capetillo, Vice-Director of Translational Research and Director of Innovation since end 2015, has been leading these key strategic areas for the CNIO, enhancing translational research at CNIO in collaboration with the CNIO Experimental Therapeutics Programme. In 2016, we leveraged on public-private partnerships in order to bring our research results closer to the patient. A project based on Dr. Djouder’s findings that boosting levels with nicotinamide riboside ( NR ) prevented and abolished aggressive tumour formation, received nearly 1 Mio in funding from MINECO’s Retos-Colaboración Programme. The aim of the project is the development of a new NR-based therapy for use in hepatocellular carcinoma and other tumours. We continue to track previously licensed programmes from our drug discovery projects led by the Experimental Therapeutics Programme ; one focused on Pim kinase inhibitors and another one on ATR inhibitors, both are very close to reaching the clinical testing stage. Commercialisation and return on investment in research remain priorities. Royalty income and milestone payments collected in 2016 raised more than 650 thousand euros. This includes revenues from patent licences as well as from commercialisation of research tools such as monoclonal antibodies. Following CNIO’s policy of royalty revenue share, this income flows back to the CNIO’s research activities as well as to the inventors themselves. A total of 44 inventors, about 10% of the researchers at CNIO, have contributed and benefited from this achievement. Industrial partnerships remain key to achieve valorisation of scientific knowledge. Valorisation is not just about ‘ money ’, but also about the impact that can be created through successful alliances with industry. In such collaborations, both the CNIO and its industrial counterpart are committed to working towards a common goal and to jointly undertaking all the steps needed to conduct the research, as well as to identifying the best possible protection and commercialisation of the results. In 2016, new contracts with industry secured future revenues for collaborative research that amount to nearly 4 million euros, which represents about 10 % of CNIO’s annual income. The CNIO External Scientific Advisory Board ( SAB ), currently chaired by Mariann Bienz, is of utmost importance for guiding the strategic plans of the CNIO as well as for the review of our research groups. We would like to wholeheartedly thank our former SAB member Joan Massagué ( Memorial Sloan Kettering Cancer Center, New York, USA ) for his committed dedication to the CNIO SAB that lasted for over a decade ( 2003-2016 ). Joan Massagué, served as Chair of the CNIO SAB from 2011 to 2014, and as member for the remaining 9 years. During 2016, we welcomed two new members to our SAB, namely Stephen Frye, Director of the Centre for Integrative Chemical Biology and Drug Discovery at the University of North Carolina Eshelman School of Pharmacy in Chapel Hill ( USA ); and Ada E. Yonath, Nobel Prize winner in Chemistry ( 2009 ) and Director of the Helen and Milton A. Kimmelman Center for Biomolecular Structure and Assembly at the Weizmann Institute of Science in Rehovot ( Israel ). I would like to take this opportunity to thank all those who helped the CNIO by sponsoring our students, postdoctoral programmes and the stays of several researchers. I hereby extend my gratitude to the Banco Santander Foundation for funding postdoctoral stays at the CNIO and the IE Business School course, the La Caixa Foundation for fostering international PhD fellowships, the Seve Ballesteros Foundation that supports the Seve-Ballesteros Foundation-CNIO Brain Tumour Group, and the Jesus Serra Foundation for supporting the Visiting Scientists Programme and the Dean’s Office. During 2016, we hosted Patrick Sung, Professor of Molecular Biophysics and Biochemistry and of SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 11 FOREWORD Therapeutic Radiology, Yale University School of Medicine in New Haven ( USA ), who started his Sabbatical at CNIO in 2015 and split his stay at the Centre in two short periods. I also wish to thank the Banc Sabadell Foundation for sponsoring a series of Distinguished Seminars at the CNIO given by ‘ outside- the-box speakers ’ who provided novel perspectives that contribute to the CNIO’s transdisciplinary environment. During 2016, we had the privilege to listen to : Romain Quidant, ICREA Research Professor at the Institute of Photonic Sciences ( ICFO ), Barcelona, Spain ; Andrés Moya, Professor at the University of Valencia, Spain ; Francisco Juan Martínez Mojica, Professor at the University of Alicante, Spain ; and Francisco J. Ayala, an evolutionary biologist and philosopher at the University of California, Irvine, USA. The Banc Sabadell Foundation also sponsored one of the WISE seminars, given by Edurne Pasaban, Mountaineer, Tolosa, Spain. Furthermore, I would like to highlight the work that is being carried out at the CNIO Women and Science ( WISE ) Office. During 2016, we had the pleasure of listening to María Concepción Ferreras, Head of You Tube Partnerships in Southern Europe and Russia ; Margery Resnick, Professor at the Massachusetts Institute of Technology in Cambridge ( USA ), and President of the International Institute in Madrid ; Pilar Garrido, Oncologist at the Instituto Ramón y Cajal de Investigación Sanitaria ( IRYCIS ), Ramón y Cajal University Hospital, Madrid ; Edurne Pasaban, Mountaineer, Tolosa, Spain ; María Teresa Fernández de la Vega, President of Women for Africa Foundation ( Fundación Mujeres por África ), and former First Deputy Prime Minister of Spain ; Tánia Balló, documentalist and film director, Barcelona, Spain ; Ángeles González-Sinde, scriptwriter, film director and former Spanish Minister of Culture ; and Christina Rosenvinge, Spanish singer-songwriter, actress and producer. Together with Belén Yuste and Sonnia L. Rivas Caballero, from Rocaviva Eventos, Madrid, we also organised and hosted the CNIO Exhibition ‘ Marie Skłodowska-Curie : a Polish girl in Paris ’. Guadalupe Martín Martín, Medical Physicist at the Hospital Universitario de Fuenlabrada, gave us the opening talk about the role of Marie’s research breaking barriers for women in science at the time and starting the field of radiotherapy. During 2016, we were also very excited to incorporate a journalist into our Communications Office ; Cristina de Martos previously worked as a journalist for the health section of El Mundo ( the second most read newspaper in Spain ) for eight years. In 2016, the CNIO appeared in the printed and digital press more than 2,600 times, thus helping to bring the CNIO to the forefront of public awareness and interweaving it into our country’s culture. Throughout the year, the featured stories received nearly 83,000 hits ( EurekAlert ! news service ) from around the world and were taken up by prestigious international media such as the BBC, The Guardian, The Scientist or Scientific American. One of the most widely commented articles in 2016 was the one authored by the researchers Manuel Serrano and Lluc Mosteiro on cell reprogramming ; it was published in November in the prestigious Science journal. Their discovery was covered by important media outlets such as radio and television, and even made it to the first page of the daily, El Mundo. The ‘ CNIO Friends ’ initiative, devoted to raising funds for cancer research at the CNIO, celebrated its first two years of existence at the end of 2016. At that time, the initiative had about 800 Friends who showed their unwavering commitment at all times. Thanks to them, we put in place the first three cancer research grants in 2016. The first two grants sponsored the researchers Paulina Gómez, from the Genetic and Molecular Epidemiology Group, and Vera Pancaldi, from the Structural Computational Biology Group. The third grant came from an agreement with the Juegaterapia Foundation ( http ://juegaterapia.org/) that is devoted to helping children affected by cancer. Thanks to this collaboration, the CNIO will hire a researcher in 2017 to investigate neuroblastoma and central nervous system tumours, two of the most common tumours in children. Businesses also joined our CNIO Friends community. In this context, the CNIO signed a collaboration agreement with ASISA Vida in relation to a new company product that includes specific coverage for gynaecological cancer. The CLH Group also participated in the sponsorship of the CNIO through an agreement aimed to support research and training of CNIO’s scientific personnel. Last but not least, I would like to thank all the CNIO volunteers who make it possible for us to move our mission forward, and of course, to the entire CNIO Friends community. Combining society’s efforts with the endeavours of the research community can make a significant difference for the future of cancer. Finally, I would like to thank all of those who have once again collaborated on the elaboration of this Annual Report, with especial thanks to Sonia Cerdá who is responsible for this CNIO publication, as well as to our collaborators : the visual artist Amparo Garrido and the underbau graphic design team. ANNUAL REPORT 2016 12 ORGANISATION OF RESEARCH MARIA A. BLASCO DIRECTOR ALFONSO VALENCIA VICE-DIRECTOR OF BASIC RESEARCH MOLECULAR ONCOLOGY PROGRAMME Manuel Serrano Programme Director Manuel Serrano Tumour Suppression Group Mariano Barbacid Experimental Oncology Group Maria A. Blasco Telomeres and Telomerase Group Marcos Malumbres Cell Division and Cancer Group Óscar Fernández-Capetillo Genomic Instability Group Ana Losada Chromosome Dynamics Group Juan Méndez DNA Replication Group María S. Soengas Melanoma Group Héctor Peinado Microenvironment and Metastasis Junior Group Manuel Valiente Brain Metastasis Junior Group Alejo Efeyan Metabolism and Cell Signalling Junior Group CNIO CANCER CELL BIOLOGY PROGRAMME Erwin F. Wagner Programme Director Erwin F. Wagner Genes, Development and Disease Group Francisco X. Real Epithelial Carcinogenesis Group Mirna Pérez-Moreno Epithelial Cell Biology Junior Group Nabil Djouder Growth Factors, Nutrients and Cancer Junior Group Massimo Squatrito Seve Ballesteros Foundation-CNIO Brain Tumour Junior Group STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME Alfonso Valencia Programme Director Alfonso Valencia Structural Computational Biology Group Daniel Lietha Cell Signalling and Adhesion Junior Group Santiago Ramón-Maiques Structural Bases of Genome Integrity Junior Group Ramón Campos-Olivas Spectroscopy and Nuclear Magnetic Resonance Unit Fátima Al-Shahrour David G. Pisano ( until May ) Bioinformatics Unit Salvador J. Capella Gutierrez ( since June ) National Bioinformatics Institute Unit Jasminka Boskovic Electron Microscopy Unit Inés Muñoz Crystallography and Protein Engineering Unit SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 13 ÓSCAR FERNÁNDEZ-CAPETILLO VICE-DIRECTOR OF TRANSLATIONAL RESEARCH HUMAN CANCER GENETICS PROGRAMME Javier Benítez Programme Director Javier Benítez Human Genetics Group Mercedes Robledo Hereditary Endocrine Cancer Group Núria Malats Genetic and Molecular Epidemiology Group Miguel Urioste Familial Cancer Clinical Unit Juan C. Cigudosa Molecular Cytogenetics and Genome Editing Unit Anna González-Neira Human Genotyping-CEGEN Unit CLINICAL RESEARCH PROGRAMME Manuel Hidalgo ( until December ) Programme Director Manuel Hidalgo ( until December ) Gastrointestinal Cancer Clinical Research Unit Miguel Quintela-Fandino Breast Cancer Junior Clinical Research Unit David Olmos Prostate Cancer Junior Clinical Research Unit Luis J. Lombardía Molecular Diagnostics Unit Joaquín Martínez-López H12O-CNIO Haematological Malignancies Clinical Research Unit Luis Paz-Ares H12O-CNIO Lung Cancer Clinical Research Unit BIOBANK Manuel M. Morente Director ÓSCAR FERNÁNDEZ-CAPETILLO DIRECTOR OF INNOVATION BIOTECHNOLOGY PROGRAMME Fernando Peláez Programme Director Orlando Domínguez Genomics Core Unit Sagrario Ortega Transgenic Mice Core Unit Giovanna Roncador Monoclonal Antibodies Core Unit Francisca Mulero Molecular Imaging Core Unit Lola Martínez Flow Cytometry Core Unit Diego Megías Confocal Microscopy Core Unit Javier Muñoz Proteomics Core Unit Alba De Martino Histopathology Core Unit Isabel Blanco Animal Facility ( Vivotecnia Management & Services ) EXPERIMENTAL THERAPEUTICS PROGRAMME Joaquín Pastor Programme Director Sonia Martínez Medicinal Chemistry Section Carmen Blanco Biology Section Susana Velasco CNIO-Lilly Cell Signalling Therapies Section María José Barrero CNIO-Lilly Epigenetics Section TECHNOLOGY TRANSFER AND VALORISATION OFFICE Anabel Sanz Director ANNUAL REPORT 2016 14 Vice-Direction of Basic Research SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 15 Molecular Oncology Programme 18 Tumour Suppression Group 20 Experimental Oncology Group 24 Telomeres and Telomerase Group 28 Cell Division and Cancer Group 32 Genomic Instability Group 36 Chromosome Dynamics Group 40 DNA Replication Group 44 Melanoma Group 48 Microenvironment & Metastasis Junior Group 52 Brain Metastasis Junior Group 54 Metabolism and Cell Signalling Junior Group 56 Cancer Cell Biology Programme 58 Genes, Development and Disease Group 60 Epithelial Carcinogenesis Group 64 Epithelial Cell Biology Junior Group 68 Growth Factors, Nutrients and Cancer Junior Group 70 Seve Ballesteros Foundation-CNIO Brain Tumour Junior Group 72 Structural Biology and Biocomputing Programme 74 Structural Computational Biology Group 76 Cell Signalling and Adhesion Junior Group 80 Structural Bases of Genome Integrity Junior Group 82 Spectroscopy and Nuclear Magnetic Resonance Unit 84 Bioinformatics Unit 86 National Bioinformatics Institute Unit 88 Electron Microscopy Unit 90 Crystallography and Protein Engineering Unit 92 ANNUAL REPORT 2016 16 ALFONSO VALENCIA Vice-Director of Basic Research SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 17 “ My main activity, as Vice-Director for Basic Research, is to work together with CNIO’s Basic Research Groups in order to enhance scientific excellence and foster collaboration.” There have been several encouraging developments this year pertaining to the organisational aspects within the CNIO’s Basic Research domain : 1 Junior Group Leader, Alejo Efeyan, joined the CNIO to conduct a new line of research in the area of metabolism and cell signalling in cancer ; the two previous Units that provided support in Bioinformatics and Translational Bioinformatics were reorganised and merged into one single Bioinformatics Unit headed by Fátima Al-shahrour ( it has already started operations in collaboration with a number of CNIO Groups ); the inclusion of CNIO’s protein production facility ( previously integrated in the Proteomics Unit ) in the Crystallography Unit, now called the Crystallography and Protein Engineering Unit ; and the recruitment of Salvador J. Capella to lead the National Bioinformatics Institute ( INB-ISCIII ) Unit. This constructive trend will be further strengthened in the coming year thanks to the creation of the Biological Text Mining Unit in the area of Computational Biology, headed by Martin Krallinger ( this new Unit will become operational at the beginning of 2017 ); and the new recruitments in strategic areas of biochemistry- structural biology and electron microscopy - protein complexes. Furthermore, the increase of collaborations with internal and external groups, as well as the consolidation of the projects developed with the Experimental Therapeutics Programme, will also lead to a further bolstering of our overall scientific output. ANNUAL REPORT 2016 18 VICE-DIRECTION OF BASIC RESEARCH MOLECULAR ONCOLOGY PROGRAMME MANUEL SERRANO Programme Director SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 19 MOLECULAR ONCOLOGY PROGRAMME It is my pleasure to introduce the highlights of the Molecular Oncology Programme in 2016. First of all, my enthusiastic and warm welcome to Alejo Efeyan, who joined the CNIO early this year to lead the Metabolism & Cell Signalling Junior Group. Alejo is a brilliant young scientist who trained as a postdoctoral fellow with David Sabatini, at the Massachusetts Institute of Technology, Cambridge. Cancer cells are metabolically hyperactive and an exciting discovery in recent years has been the realisation that cancer cells have mutations in the pathways that detect nutrient availability. Understanding how the nutrient sensing mechanisms contribute to cancer is the main goal of the Metabolism & Cell Signalling Junior Group. Alejo’s outstanding career and his original project have been awarded with a prestigious and generous grant from the European Research Council. His Group is now settled and fully operative. We are all very proud of having him here with us and we wish him all the best ! It is also very gratifying that the two other Junior Groups that joined the CNIO during 2015 have continued to successfully consolidate their teams and their projects throughout 2016. The Brain Metastasis Junior Group, led by Manuel Valiente, now has promising candidate small compounds that inhibit metastasis initiation in brain slices. Likewise, the Microenvironment & Metastasis Junior Group, led by Héctor Peinado is making impressive progress towards detecting how the vesicles shed by tumours ( known as exosomes ) are distributed throughout the organism, modifying it, and thereby making it more receptive for metastatic seeding. In the following pages, you will read about several surprising discoveries that expand our understanding of cancer and that might pinpoint new therapeutic strategies in the near future. For example, a protein that regulates mRNA stability, CEBP4, and contributes to cancer progression ( Melanoma Group, Nat. Commun. 2016 ); or telomere-derived transcripts that play a key role in chromosomal integrity through the stabilisation of telomeres ( Telomeres & Telomerase Group, Nat. Commun. 2016 ). Other discoveries concern basic mechanisms that are altered in cancer. For example, it has been found that the infliction of damage to tissues triggers cell plasticity in the surrounding cells and that cytokine IL-6 is a key factor in this process ( Tumour Suppression Group, Science 2016 ). You will also read about a rather unique DNA polymerase named PrimPol ( DNA Replication Group ); about how cohesins regulate transcription, an unsuspected role for these proteins traditionally involved in sister chromatid cohesion ( Chromosome Dynamics Group ); and about a kinase named MASTL that is upregulated in cancer and its inhibition blocks the proliferation of some cancer cells ( Cell Division & Cancer Group ). The identification of preclinical anti-cancer treatments is the ultimate goal of the Molecular Oncology Programme. In this regard, we are proud of two relevant contributions : a novel therapeutic approach for lung adenocarcinoma based on two inhibitory molecules that are effective even against aggressive cancers lacking p53 ( Experimental Oncology Group, Nat. Med. 2016 ); and the identification of compounds that block DNA repair with therapeutic efficacy against acute myeloid leukaemia ( Genomic Instability Group, Sci. Signal. 2016 ). “ Congratulations to Marcos Malumbres and Óscar Fernández-Capetillo for being elected as members of the European Molecular Biology Organization ( EMBO ).” ANNUAL REPORT 2016 20 VICE-DIRECTION OF BASIC RESEARCH TUMOUR SUPPRESSION GROUP Manuel Serrano Group Leader Staff Scientists Susana Llanos, Bárbara Martínez, Daniel Muñoz ( until August ), Cristina Pantoja SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 21 MOLECULAR ONCOLOGY PROGRAMME | TUMOUR SUPPRESSION GROUP OVERVIEW Tumour suppressors are genes that can prevent the development of cancer. All our cells have a functional set of these genes, but they can become defective over time. The affected cells thus become partially unprotected and, combined with additional mutations in other genes, can give rise to cancer. Understanding how tumour suppressor genes work may help us to design drugs that block cancer. Tumour suppressor genes are now known to control many aspects of cell biology and organismal physiology, such as cellular pluripotency, cell senescence, and metabolism. We aim to achieve an integrated understanding of cancer protection. Our goals are to : ɗ Understand the mechanisms of tumour suppression and identify new tumour suppressor regulators. ɗ Study the interplay between tumour suppression and ageing. ɗ Analyse the involvement of tumour suppressors in the regulation of metabolism and protection from metabolic damage. ɗ Characterise cellular senescence as a tumour suppression mechanism. ɗ Investigate cellular pluripotency and the involvement of tumour suppressors in the process of reprogramming to induced pluripotent stem ( iPS ) cells. ɗ Explore the role of cell plasticity in cancer, tissue regeneration, and ageing. ɗ Search for new frontiers in cell plasticity. “ We have found that damaged cells secrete factors that promote reparative activities in the surrounding cells, including loss of differentiation and plasticity. This could be beneficial to repair the damaged tissues, but it could also favour the expansion of dormant cancer cells.” Post-Doctoral Fellows Timothy Cash, Cian J. Lynch, Gianluca Varetti Graduate Students Noelia Alcázar, Raquel Bernad, Selim Chaib, Dafni Chondronasiou, Elena López-Guadamillas ( until September ), Lluc Mosteiro, Miguel Rovira Technician Maribel Muñoz ( TS )* *Titulado Superior ( Advanced Degree ) Student in practice Isabel Calvo ANNUAL REPORT 2016 22 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS New tumour markers for the prognosis of head and neck cancer Head and neck cancers include a heterogeneous group of tumours located in the oral cavity, pharynx and larynx. The survival rate of patients with this pathology has hardly improved over the last decade. Stratification of patients has been limited, until now, to a clinical classification and not a molecular one. Analysis of patients ’ biopsies showed that about half of them possess high levels of the p21 protein as well as mTOR activation. We have unravelled the molecular mechanism by which p21 levels are linked to the activity of mTOR ( FIGURE 1 ). When the mTOR protein is inactive, it dictates the degradation of p21, and, conversely, when mTOR is active, p21 becomes stable. The presence of both markers, active mTOR and high levels of p21, predict a less aggressive evolution of the disease. This may help in choosing from amongst different therapeutic options for these patients. Antioxidant defences delay ageing and age-related diseases Accumulation of cell damage plays an important role in ageing. There is no clear answer about which types of cellular damage Figure 1 Model of mTOR-mediated regulation of p21. Non-phosphorylated 4E-BP1 binds to p21 and promotes p21 degradation, and this is associated to more aggressive cancers. 4E-BP1 phosphorylation by mTOR disrupts the 4E-BP1/p21 complex ensuing stable p21 accumulation, and this is associated to less aggressive cancers. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 23 MOLECULAR ONCOLOGY PROGRAMME | TUMOUR SUPPRESSION GROUP ∞ PUBLICATIONS ∞ Mosteiro L, Pantoja C, Alcazar N, Marión RM, Chondronasiou D, Rovira M, Fernan- dez-Marcos PJ, Muñoz-Martin M, Blan- co-Aparicio C, Pastor J, Gómez-López G, de Martino A, Blasco MA, Abad M, Serrano M ( 2016 ). Tissue damage and senescence provide critical signals for cellular reprogramming in vivo. Science 354, 6315. pii : aaf4445. ∞ Ambrogio C, Gómez-López G, Falcone M, Vidal A, Nadal E, Crosetto N, Blasco RB, Fernández-Marcos PJ, Sánchez-Céspedes M, Ren X, Wang Z, Ding K, Hidalgo M, Serrano M, Villanueva A, Santamaría D, Barbacid M ( 2016 ). Combined inhibition of DDR1 and Notch signaling is a ther- apeutic strategy for KRAS-driven lung adenocarcinoma. Nat Med 22, 270-277. ∞ Llanos S, Serrano M ( 2016 ). Senescence and cancer : in the name of immunosup- pression. Cancer Cell 30, 507-508. ∞ Fernandez-Marcos PJ, Serrano M ( 2016 ). Mitochondrial damage induces senes- cence with a twisted arm. Cell Metab 23, 229-230. ∞ Knafo S, Sánchez-Puelles C, Palomer E, Delgado I, Draffin JE, Mingo J, Wahle T, Kaleka K, Mou L, Pereda-Perez I, Klosi E, Faber EB, Chapman HM, Lozano-Montes L, Ortega-Molina A, Ordóñez-Gutiérrez L, Wandosell F, Viña J, Dotti CG, Hall RA, Pulido R, Gerges NZ, Chan AM, Spaller MR, Serrano M, Venero C, Esteban JA. ( 2016 ). PTEN recruitment controls synaptic and cognitive function in Alzheimer’s models. Nat Neurosci 19, 443-453. ∞ Nóbrega-Pereira S, Fernandez-Marcos PJ, Brioche T, Gomez-Cabrera MC, Salva- dor-Pascual A, Flores JM, Viña J, Serrano M ( 2016 ). G6PD protects from oxidative damage and improves healthspan in mice. Nat Commun 7, 10894. ∞ Llanos S, García-Pedrero JM, Mor- gado-Palacin L, Rodrigo JP, Serra- no M ( 2016 ). Stabilization of p21 by mTORC1/4E-BP1 predicts clinical outcome of head and neck cancers. Nat Commun 7, 10438. ∞ López-Guadamillas E, Muñoz-Martin M, Martinez S, Pastor J, Fernandez-Marcos PJ, Serrano M ( 2016 ). PI3Ka inhibition re- duces obesity in mice. Aging 8, 2747-2753. ∞ Casas-Tintó S, Maraver A, Serrano M, Fer- rús A ( 2016 ). Troponin-I enhances and is required for oncogenic overgrowth. Oncotarget 7, 52631-52642. ∞ García-Carpizo V, Sarmentero J, Han B, Graña O, Ruiz-Llorente S, Pisano DG, Ser- rano M, Brooks HB, Campbell RM, Barrero MJ ( 2016 ). NSD2 contributes to oncogenic RAS-driven transcription in lung cancer cells through long-range epigenetic ac- tivation. Sci Rep 8, 32952. ∞ López-Guadamillas E, Fernández-Marcos PJ, Pantoja C, Muñoz-Martin M, Martínez D, Gómez-López G, Campos-Olivas R, Valverde AM, Serrano M ( 2016 ). p21Cip 1 plays a critical role in the physiological adaptation to fasting through activation of PPARa. Sci Rep 6, 34542. ∞ Serrano M ( 2016 ). Unraveling the links between cancer and aging. Carcinogen- esis 37, 107. ∞ Llanos S, García-Pedrero JM ( 2016 ). A new mechanism of regulation of p21 by the mTORC1/4E-BP1 pathway predicts clinical outcome of head and neck cancer. Mol Cell Oncol 3, e1159275. are more relevant for ageing. Although the accumulation of oxidative damage with ageing is undisputed, the large majority of attempts to prove that oxidative damage is relevant for ageing have failed. All these attempts, however, have manipulated only one component of the complex network of antioxidant defences. In contrast to these previous attempts, we have approached this issue by increasing the levels of NADPH, a simple co-factor required for almost all antioxidant reactions and whose levels are known to determine the global antioxidant capacity of cells. To achieve this, we generated transgenic mice with an increased expression throughout their bodies of glucose-6-phosphate dehydrogenase ( G6PD ), one of the most important enzymes for the production of NADPH. We found that G6PD transgenic mice have overall higher levels of NADPH and, consequently, a better protection against oxidative damages. Importantly, these mice are not predisposed to cancer and, indeed, have a modest increase in longevity. These observations point to a novel strategy to delay ageing-related diseases, including cancer. Senescent cells provide critical signals for cellular reprogramming The mechanisms involved in the reprogramming of differentiated cells inside a living organism remain to be elucidated. Senescence is a cellular response to damage characterised by an abundant production of cytokines and other secreted factors that, together with the recruitment of inflammatory cells, results in tissue remodelling. We have shown that in vivo expression of the reprogramming factors OCT4, SOX2, KLF4 and cMYC ( OSKM ) triggers two divergent cellular outcomes : most cells undergo senescence, while other cells undergo reprogramming, both occurring in close physical association. OSKM-induced senescence requires the tumour suppressor locus Ink4a/Arf, which, via the production of the cytokine IL6, creates an optimal tissue environment for in vivo reprogramming. We concluded that tissue injury or ageing, through cellular senescence and cytokine IL6, favour in vivo reprogramming by OSKM ( FIGURE 2 ). These findings could be relevant for tissue repair and open new strategies to manipulate reprogramming in vivo. s Figure 2 Tissue damage and ageing favour in vivo reprogramming. Expression of OSKM in vivo induces the reprogramming of a small population of cells, as well as damage and senescence in many other cells. Senescent cells release factors that promote the reprogramming of neighbouring cells, and IL6 is a critical mediator. Tissue injury and ageing also favour in vivo reprogramming through the accumulation of senescent cells. ANNUAL REPORT 2016 24 VICE-DIRECTION OF BASIC RESEARCH EXPERIMENTAL ONCOLOGY GROUP Mariano Barbacid Group Leader Staff Scientists Matthias Drosten, Carmen Guerra, Monica A. Musteanu, David Santamaría ( until August ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 25 MOLECULAR ONCOLOGY PROGRAMME | EXPERIMENTAL ONCOLOGY GROUP OVERVIEW KRAS oncogenes have been implicated in about one fifth of all human cancers including lung and pancreatic adenocarcinomas, 2 of the tumour types with the worst prognosis. Unfortunately, identification of suitable therapies to treat these tumours remains elusive and patients are still treated with cytotoxic compounds approved over 2 decades ago. The recent discovery that these tumours display intra-tumour heterogeneity adds another layer of complexity that needs to be addressed. Hence, our laboratory has decided to search for novel therapeutic targets that may contribute to the early stages of lung tumour development, hoping that these targets will be present in all tumour cells – including cancer initiating cells and cancer stem cells – and not only in limited populations of evolving clones. In addition, we have continued our quest to validate known targets ( mainly those of the MAPK and PI3K pathways ) using genetically engineered mouse tumour models with the ultimate goal of establishing rational combination therapies that may provide significant therapeutic benefits in the clinic. Significance ɗ We have shown that human lung tumours respond efficiently to combinations of DDR1 and NOTCH inhibitors in PDX models. ɗ We have provided a mechanistic explanation for the exclusive presence of K-RAS or EGFR mutations in human lung adenocarcinomas. ɗ We have demonstrated that the different incidence of H-RAS and K-RAS oncogenes in human tumours is due to the signalling intensity of their respective oncoproteins. Post-Doctoral Fellows Chiara Ambrogio ( until April ), Raquel García, Harrys K. C. Jacob Graduate Students M. Teresa Blasco, Laura De Esteban, Magdolna Djurec, Fernando Fernández ( since October ), Jing Li ( since December ), Laura Martín ( since September ), Marina Salmón ( since October ), Manuel Sanclemente, Lucía Simón ( until December ) Visiting Graduate Student Lavinia Cabras ( Università di Cagliari, Italy ) ( since September ) Technicians M. Carmen González ( TS )*, Silvia Jiménez ( since August ), Marta San Román. Raquel Villar *Titulado Superior ( Advanced Degree ) External Associates Alfredo Carrato ( Hospital Ramón y Cajal, Madrid ), Bruno Sainz ( Universidad Autónoma de Madrid ), Juan Velasco ( Lilly Spain ) ANNUAL REPORT 2016 26 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS Identification of novel therapeutic targets for the treatment of K-Ras driven lung adenocarcinoma The recent discovery that lung tumours display significant levels of clonal heterogeneity ( Govindan, Science, 2014 ) implies that effective therapies must target early oncogenic events/alterations present in all tumour cells and not only in clonal variants that appear during tumour development. To provide potential solutions to this key issue we decided to search for novel therapeutic targets present in the earliest stages of lung tumour development, expecting that such targets will be present in the entire tumour population including the putative cancer initiating/stem cells. Among the most highly expressed druggable genes we identified Ddr1, a locus that encodes a tyrosine protein kinase receptor. As reported early this year ( Ambrogio et al., Nat Med, 2016 ), genetic and pharmacological inhibition of Ddr1 prevented progression of K-Ras driven p53 wild type, but not p53 mutant tumours. Yet concomitant inhibition of Ddr1 and Notch, a downstream mediator of Ddr1 activity, led to a significant anti-tumour effect even in aggressive K-RasG 12V ; p53 mutant adenocarcinomas. More importantly, this treatment induced regression of K-RAS ;p53 mutant patient-derived lung ortho- xenografts ( PDX ) with a therapeutic efficacy superior to standard chemotherapy. Identification of additional targets present in these early K-Ras mutant driven lung cells should expand the therapeutic opportunities to treat K-RAS mutant tumours in the clinic, thus by-passing the challenges derived from the development of intra-tumour heterogeneity. Lack of selective advantage for lung cells expressing K-RAS and EGFR oncogenes Activating mutations in KRAS and EGFR, the 2 most frequent oncogenic drivers in human lung adenocarcinoma, occur in a mutually exclusive manner suggesting functional redundancy and implying lack of positive selection. By means of a mouse model engineered to induce expression of mutant EGFRL 858R in advanced tumours driven by a resident KrasG 12V oncogene, we show that, instead, their co-expression is detrimental for the progression of lung adenocarcinoma. In vivo expression of EGFRL 858R in KrasG 12V-driven tumours triggers an immediate response with hallmarks of replicative stress resulting in apoptosis. Yet, a fraction of tumour cells survive, but enter a transient cytostatic state incompatible with tumour development that is fully reversible upon discontinuation of EGFRL 858R expression. Ultimately, continuous co-expression of both mutants results in the attenuation of the overall oncogenic signalling to levels compatible with cell proliferation and tumour growth. In sum, our results indicate that the mutual exclusivity of KRAS and EGFR activating mutations occurs as a combination of cellular toxicity and signal adjustment that results in the lack of selective advantage for those cells expressing both oncogenes. Whereas the wild type H-Ras and K-Ras proteins are bioequivalent, their oncogenic isoforms H-RasG 12V and K-RasG 12V induce different tumour spectra We have provided genetic evidence demonstrating that the H-Ras and K-Ras proteins are fully bioequivalent in mice. Previous studies have shown that replacement of the K-Ras alleles by H-Ras coding sequences resulted in viable mice ( Potenza et al., EMBO Rep, 2005 ). Yet, these mice displayed cardiovascular defects. Now, we have shown that these defects were due to the presence of the 4 H-Ras expressing alleles in these mice. Ablation of the 2 endogenous H-Ras alleles, hence generating mice that only express the H-Ras protein from the 2 targeted K-Ras alleles, is absolutely normal. These results appear to be at variance with the well-established observation that H-RAS and K-RAS oncogenes are involved in different human tumour types. To determine whether the oncogenic versions of the H-Ras and K-Ras proteins are also bioequivalent, we knocked-in H-RasG 12V oncogene sequences into the K-Ras locus. Germline expression of H-RasG 12V or K-RasG 12V from the K-Ras locus resulted in equal embryonic lethality. However, their expression in adult mice led to different tumour phenotypes. Whereas H-RasG 12V elicited papillomas and haematopoietic tumours, K-RasG 12V induced lung tumours and gastric lesions. The reason why H-RasG 12V expression failed to cause lung tumours is due to the induction of a senescence-like state due to excessive MAP kinase signalling. Likewise, H-RasG 12V but not K-RasG 12V induced oncogene-induced senescence in mouse embryonic fibroblasts ( MEFs ). Label-free quantitative analysis revealed that minor differences in H-RasG 12V expression levels led to drastically different biological outputs, suggesting that subtle differences in MAP kinase signalling influence the differential tumour spectra induced by RAS oncoproteins. s SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 27 MOLECULAR ONCOLOGY PROGRAMME | EXPERIMENTAL ONCOLOGY GROUP ∞ PUBLICATIONS ∞ Ambrogio C, Gómez-López G, Falcone M, Vidal A, Nadal E, Crosetto N, Blasco RB, Fernández-Marcos PJ, Sánchez-Céspedes M, Ren X, Wang Z, Ding K, Hidalgo M, Serrano M, Villanueva A, Santamaría D, Barbacid M ( 2016 ). Combined inhibition of DDR1 and Notch signaling is a ther- apeutic strategy for KRAS-driven lung adenocarcinoma. Nat Med 22, 270-277. ∞ Drosten M, Simón-Carrasco L, Hernán- dez-Porras I, Lechuga CG, Blasco MT, Ja- cob HK, Fabbiano S, Potenza N, Bustelo XR, Guerra C, Barbacid M ( 2016 ). H-Ras and K-Ras oncoproteins induce different tumor spectra when driven by the same regulatory sequences. Cancer Res. PMID : 27872088. ∞ Ambrogio C, Barbacid M, Santamaría D ( 2016 ). In vivo oncogenic conflicto trig- gered by co-existing KRAS and EGFR acti- vating mutations in lung adenocarcinoma. Oncogene. PMID : 27775074. ∞ Hernández-Porras I, Schuhmacher AJ, Garcia-Medina R, Jiménez B, Cañame- ro M, de Martino A, Guerra C ( 2016 ). K-Ras( V14I ) -induced Noonan syndrome predisposes to tumour development in mice. J Pathol 239, 206-217. ∞ Lopez S, Voisset E, Tisserand JC, Mos- ca C, Prebet T, Santamaria D, Dubreuil P, De Sepulveda P ( 2016 ). An essential pathway links FLT3-ITD, HCK and CDK6 in acute myeloid leukemia. Oncotarget 7, 51163-51173. ∞ Drosten M, Barbacid M ( 2016 ). Modeling K-Ras-driven lung adenocarcinoma in mice : preclinical validation of therapeutic targets. J Mol Med 94, 121-135. ∞ Ambrogio C, Nadal E, Villanueva A, Gómez-López G, Cash TP, Barbacid M, Santamaría D ( 2016 ). KRAS-driven lung adenocarcinoma : combined DDR1/Notch inhibition as an effective therapy. ESMO Open 1, e000076. ∞ AWARDS AND RECOGNITION ∞ Advanced Grant of the European Research Council ( 2016-2021 ). ∞ Keynote Speaker, 24th congress of the Eu- ropean Association for Cancer Research, Manchester, UK. ∞ Keynote Speaker, International Conference on Predictive Cancer Models, Barcelona, Spain. ∞ Keynote Speaker, 49th Annual Meeting of the European Society of Human Genetics, Barcelona, Spain. ∞ Meeting Organiser and Session Chair, 28th Pezcoller Symposium, Trento, Italy. ∞ Honorary Member, Royal Academy of Medicine of Valencia, Spain. Figure H-RasG 12V but not K-RasG 12V expressed from the K-Ras locus induces senescence. ( A ) Growth curve of K-RasG 12V ( KV 12 ) and H-RasG 12V ( KHRasV 12 ) expressing MEFs. ( B ) Representative images of senescence-associated SA-βGal staining in these MEFs. ( C ) E1A cooperated with H-RasG 12V, but not with K-RasG 12V, to transform immortal MEFs. A B C ANNUAL REPORT 2016 28 VICE-DIRECTION OF BASIC RESEARCH TELOMERES AND TELOMERASE GROUP Maria A. Blasco Group Leader Staff Scientists Isabel López de Silanes, Rosa M. Marión, Paula Martínez, Marinela Méndez, Águeda M. Tejera, Elisa Varela ( until May ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 29 MOLECULAR ONCOLOGY PROGRAMME | TELOMERES AND TELOMERASE GROUP OVERVIEW We study the mechanisms by which tumour cells are immortal and normal cells are mortal. Immortality is one of the most universal characteristics of cancer cells. The enzyme telomerase is present in more than 95% of all types of human cancers and absent in normal cells in the body. Telomeres are nucleoprotein complexes located at the ends of chromosomes, essential for chromosome protection and genomic stability. Progressive shortening of telomeres associated with organism ageing leads to ageing. When telomeres are altered adult stem cells have a maimed regenerative capacity. Our research aims are : ɗ Generating mouse models to validate telomeres and telomerase as therapeutic targets for cancer and age-related diseases. ɗ Deciphering the interplay between telomeres and DNA repair pathways. ɗ Studying the role and regulation of non-coding telomeric RNAs or TERRA. ɗ Testing telomerase gene therapy in ‘ telomere syndromes ’ and age-related diseases. ɗ Elucidating the role of telomerase and telomeres in adult stem cell biology and in nuclear reprogramming of differentiated cells to iPS cells. “ We have demonstrated that TERRA long non-coding RNAs are essential for telomere protection.” Post-Doctoral Fellow Kurt Whittemore Graduate Students Leire Bejarano, Aksinya Derevyanko, Lole Ferrara, Juan José Montero, Miguel Ángel Muñoz, Juan Manuel Povedano ( until June ) Technicians Rosa M. Serrano, Nora Soberón ( until June ) ( TS )* *Titulado Superior ( Advanced Degree ) Student in practice Ana C. Moises da Silva ANNUAL REPORT 2016 30 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS Fighting aplasic anaemia using a therapy designed to delay ageing Aplasic anaemia is a rare, potentially fatal disease of the blood, by which the bone marrow is unable to generate blood cells at the appropriate pace. The disease can be hereditary or acquired and develops at any stage of life. A subgroup of the inherited form is caused by replicative impairment of haematopoietic stem and progenitor cells owing to very short telomeres due to mutations in telomerase and other telomere components. An abnormal telomere shortening is also described in cases of acquired aplasic anaemia. We have tested the efficacy of our telomerase gene therapy, originally designed to delay ageing, in two independent mouse models of aplasic anaemia due to short telomeres. We found that a high dose targets the bone marrow compartment, including haematopoietic stem cells. Telomerase treatment following telomere attrition in bone marrow cells rescues aplasic anaemia and mouse survival. Improved survival is associated with a significant increase in telomere length in peripheral blood and bone marrow cells, as well as improved blood counts. Our telomerase gene therapy represents a novel therapeutic strategy to treat aplasic anaemia provoked or associated with short telomeres. Mice with hyper-long telomeres and unaltered genes Telomere length is genetically determined, but in the past we were able to generate mouse embryonic stem ( ES ) cells with telomeres twice the size of normal ones. We have now used such ES cells with ‘ hyper-long ’ telomeres, traceable thanks to the co-expression of green fluorescent protein ( GFP ), to generate chimaeric mice containing cells with both hyper-long and normal telomeres. We showed that chimaeric mice contain GFP-positive cells – bearing hyper-long telomeres – in all mouse tissues ( FIGURE 1 ), display normal tissue histology, as well as normal survival. Both hyper-long and normal telomeres shorten with age, but GFP-positive cells manage to retain longer telomeres as the mice age. These chimaeric mice also accumulate fewer cells with short telomeres and less DNA damage with age, and express lower levels of p53. Cells with hyper-long telomeres are longitudinally maintained or enriched with age in highly renewing compartments ( i.e. blood ). We demonstrated that mice with functional, longer and better preserved telomeres can be generated without the need for genetic manipulations, such as telomerase overexpression. Telomeric RNAs are essential to maintain telomeres Despite their especially compact structure, which is difficult to access, telomeres transcribe information like the rest of the DNA generating long non-coding RNAs known as TERRA. Deciphering the role of TERRA was one of the unsolved issues of telomere biology in the past decade. This was, in part, due to a lack of knowledge on the TERRA loci, which had prevented functional genetic studies. We had already shown that mouse TERRA arise mainly from the subtelomere of chromosome 18 and to a lesser extent from the subtelomere of chromosome 9. We have now described that long non-coding RNAs with TERRA features are Figure 1 Representative image of an eye of chimaeric mice. Cells bearing hyper-long telomeres are visualised in green. Telomeres appear in red. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 31 MOLECULAR ONCOLOGY PROGRAMME | TELOMERES AND TELOMERASE GROUP ∞ PUBLICATIONS ∞ Mosteiro L, Pantoja C, Alcázar N, Marión RM, Chondronasiou D, Rovira M, Fernán- dez-Marcos PJ, Muñoz-Martin M, Blan- co-Aparicio C, Pastor J, Gómez-López G. de Martino A, Blasco MA, Abad M, Serrano M ( 2016 ). Tissue damage and senescence provide critical signals for cellular reprogramming in vivo. Science 354, 6315. pii : aaf4445. ∞ Fernández-Alvira JM, Fuster V, Dorado B, Soberón N, Flores I, Gallardo M, Pocock S, Blasco MA, Andrés V ( 2016 ). Short tel- omere load, telomere length, and subclini- cal atherosclerosis : The PESA Study. J Am Coll Cardiol 67, 2467-2476. ∞ Montero JJ, López de Silanes I, Graña O, Blasco MA ( 2016 ). Telomeric RNAs are essential to maintain telomeres. Nat Commun 7, 12534. ∞ Varela E, Muñoz-Lorente MA, Tejera AM, Ortega S, Blasco MA ( 2016 ). Generation of mice with longer and better preserved telomeres in the absence of genetic ma- nipulations. Nat Commun 7, 11739. ∞ Bär C, Povedano JM, Serrano R, Ben- itez-Buelga C, Popkes M, Formentini I, Bobadilla M, Bosch F, Blasco MA ( 2016 ). Telomerase gene therapy rescues tel- omere length, bone marrow aplasia and survival in mice with aplasic anaemia. Blood 127, 1770-1779. ∞ Aguado T, Gutiérrez FJ, Aix E, Schneider RP, Giovinnazo G, Blasco MA, Flores I ( 2016 ). Telomere length defines the car- diomyocyte differentiation potency of mouse induced pluripotent stem cells. Stem Cells. PMID : 27612935. ∞ Benitez-Buelga C, Vaclová T, Ferreira S, Urioste M, Inglada-Perez L, Soberón, N, Blasco MA, Osorio A, Benitez J ( 2016 ). Molecular insights into the OGG1 gene, a cancer risk modifier in BRCA1 and BRCA2 mutations carriers. Oncotarget 7, 25815- 25825. ∞ Martínez P, Gómez-López G, Pisano DG, Flores JM, Blasco MA ( 2016 ). A genetic interaction between RAP1 and telomerase reveals an unanticipated role for RAP1 in telomere maintenance. Aging Cell. PMID : 27586969. ∞ Boskovic J, Martínez-Gago J, Mendez-Per- tuz M, Buscato A, Martínez-Torrecuadrada JL, Blasco MA ( 2016 ). Molecular Archi- tecture of Full Length TRF1 Favors its Interaction with DNA. J Biol Chem 291, 21829-21835. ∞ Comino-Méndez I, Tejera ÁM, Cur- rás-Freixes M, Remacha L, Gonzalvo P, Tonda R, Letón R, Blasco MA, Robledo M, Cascón A ( 2016 ). ATRX driver mutation in a composite malignant pheochromocyto- ma. Cancer Genet 209,272-277. ∞ Bär C, Blasco MA ( 2016 ). Telomeres and tel- omerase as therapeutic targets to prevent and treat age-related diseases. F1000Re- search 5, pii : F1000 Faculty Rev-89. ∞ PATENTS ∞ Blasco MA, Bernardes B, Bosch F, Ayu- so E ( 2016 ). Telomerase reverse tran- scriptase for protection against aging. EP2402038B1. ∞ Flores I, Canela A, Blasco MA ( 2016 ). Methods for the determination of tel- omere length in a semi-automatic manner of every single cell in an immobilized cell population. CA2723950. ∞ Blasco MA, Bernardes de Jesus B, Baer C, Serrano Ruiz MR, Bosch I Tubert F, Ayuso E, Formentini I, Bobadilla M, Miz- rahi J ( 2016 ). Telomerase reverse tran- scriptase-based therapies for treatment of conditions associated with myocardial infarction. WO/2016/020346. ∞ Bobadilla M, Formentini I, Blasco MA, Baer C, Bosch I Tubert F ( 2016 ). Telomerase reverse transcriptase-based therapies. WO/2016/020345. ∞ AWARDS AND RECOGNITION ∞ Miguel Catalán Career Achievement Award, Regional Government of Madrid. ∞ Scientific Committee Member, Innovative Medicines Initiative 2 ( IMI2 ) Joint Under- taking, Brussels, Belgium. ∞ Member of the External Scientific Board of Instituto de Investigación Sanitaria y Biomédica de Alicante ( ISABIAL ), Ali- cante, Spain. ∞ Member, Alumni Advisory Board, Univer- sidad Autónoma de Madrid. ∞ Founding Editor, Cell Stress. ∞ Editorial Board Member, Nutrition & Healthy Aging. transcribed from the human 20q and Xp subtelomeres. We used the CRISPR-Cas9 technology to delete the 20q locus, which resulted in a dramatic decrease in TERRA levels. The deletion of the Xp locus, on the contrary, does not lead to decreased TERRA levels. These findings demonstrate that, although human TERRA arise from two loci, the 20q locus is the main origin of human TERRA. Thus, both in mice and in humans, TERRA arise from one, or at most two loci ( FIGURE 2 ). Ablation of 20q-TERRA in human cells results in a dramatic loss of telomere sequences and in the induction of a massive DNA damage response. These latter findings represent the first demonstration, in any organism, of the essential role of TERRA in the maintenance of telomeres. s Figure 2 The genomic origin of telomeric RNAs ( TERRA ). Both in mice and in humans, TERRA arise from one or at most two loci. ANNUAL REPORT 2016 32 VICE-DIRECTION OF BASIC RESEARCH CELL DIVISION AND CANCER GROUP Marcos Malumbres Group Leader Staff Scientists Mónica Álvarez, Guillermo de Cárcer, Ignacio Pérez de Castro ( until February ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 33 MOLECULAR ONCOLOGY PROGRAMME | CELL DIVISION AND CANCER GROUP OVERVIEW The Cell Division and Cancer Group is interested in deciphering the mechanisms by which cell division and cell proliferation are regulated. During the past few years, we have used different mouse models to understand the relevance of cell cycle regulators, including cell cycle kinases and phosphatases, as well as proteins involved in ubiquitin-dependent degradation, in the control of cell division and tissue physiology. Our interests are : i ) to understand the basic control mechanisms that regulate the cell division cycle ; ii ) to characterise the physiological and therapeutic consequences of cell cycle deregulation ; iii ) understanding the function of microRNAs in cell biology and tumour development, and iv ) to understand how progenitor cells and cancer stem cells control their self-renewal and proliferative properties. As a final goal, we aim to generate information that may be useful for improving therapeutic strategies against cancer cell proliferation. “ In 2016, we investigated the relevance of several mitotic regulators during cancer progression and therapy, with special focus on kinases that are currently under preclinical and clinical evaluation.” Post-Doctoral Fellows Begoña Hurtado, Carolina Maestre, María Salazar Graduate Students Ana F. Batalha Martins, María Maroto, Diego Martínez, Belén Sanz, María Sanz Technicians Aicha El Bakkali ( since March ) ( PEJ )*, María Guirola ( since March ) ( TS )**, Elisabet Zapatero ( TS )** *Plan de Empleo Joven ( Youth Employment Plan ) **Titulado Superior ( Advanced Degree ) ANNUAL REPORT 2016 34 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS Controlling the proper number of cell divisions The mammalian cell cycle is regulated by at least 2 families of inhibitors, the INK4 and Cip/Kip proteins. While elimination of individual members of these families is a frequent finding in human cancer, the consequences of eliminating this inhibitory mechanism in mammalian cells have not yet been explored. Using a combination of mutant alleles in the mouse, we have now observed that a major physiological function of cell cycle inhibitors is to prevent replicative stress. In a mouse model insensitive to INK4 proteins and deficient in p21Cip 1 and p27Kip 1, we observed that these inhibitors prevent the accumulation of DNA damage due to replicative stress in different tissues including the nervous system. Ablation of these inhibitors prevents mouse development. This effect is most likely due to hyperactivation of cyclin-dependent kinases as the replicative stress can be prevented by slightly inhibiting the enzymatic activity of these proteins ( Quereda et al., 2016 ). Cell cycle kinases as new targets for cancer therapy Cell cycle progression is controlled by phosphorylation events and cell cycle kinases are currently the focus of multiple therapeutic strategies. Inhibitors of the Aurora and Polo-like kinases are evaluated in clinical trials with promising results, at least in haematopoietic malignancies. Over the last few years, we have generated mouse models with specific mutations in these kinases in order to understand their roles in different tissues and cell types. Our recent data have uncovered an unexpected function of Polo-like kinase 1 ( Plk1 ) in the cardiovascular system ; a role that we are studying in detail in order to understand possible toxicities derived from the use of Plk1 inhibitors in patients. A relatively new serine/threonine kinase, known as MASTL ( or Greatwall in flies and Xenopus ), has been characterised as a critical node in cell division. We have previously shown that MASTL is essential for mouse embryonic development and cell cycle progression ( FIGURE ). This is due to mitotic collapse after nuclear envelop breakdown ( NEB ). MASTL is exported from the nucleus to the cytoplasm in a CRM1-dependent manner before NEB. Once at the cytoplasm, Greatwall inhibits the PP2A-B55 complexes to maintain the mitotic state. These findings have therapeutic implications since MASTL acts by blocking the function of the PP2A phosphatase, a tumour suppressor frequently altered in human cancer. This implies that the inhibition of MASTL could, at the same time, slow down cell division and reactivate tumour suppressor PP2A, a protein capable of inhibiting many of the oncogenic molecular pathways involved in cancer development. Over the past few months, we have tested this hypothesis by studying the relevance of MASTL in tumour cell proliferation and its possible use as a cancer target. In collaboration with Miguel Quintela’s Group at the CNIO and Carlos Caldas at Cancer Research UK, we analysed MASTL expression in breast cancer. Our data suggest that this protein is overexpressed in a significant number of hormone-positive and -negative tumours and correlates with poor prognosis. In collaboration with researchers at Pfizer, we used different RNAi and CRISPR techniques to analyse the effect of MASTL knockdown or knockout in breast cancer cells both in vitro and in vivo. These data indicate that some breast cancer cells require MASTL kinase activity for proliferating, suggesting that a subset of breast tumours may benefit from strategies aimed at inhibiting this kinase. We are currently studying the consequences of inhibiting MASTL in the activity of the PP2A phosphatase. Since MASTL specifically inhibits PP2A-B55 complexes, we are also characterising the relevance of the B55 family members present in the human genome. s SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 35 MOLECULAR ONCOLOGY PROGRAMME | CELL DIVISION AND CANCER GROUP Figure Schematic representation of the control of the mammalian cell cycle by cyclin-dependent kinases ( Cdks ) and MASTL. MASTL phosphorylates 2 small proteins, known as Ensa and Arpp19, which are both PP2A-B55 inhibitors. The proper balance between cell cycle kinases and phosphatases is crucial, not only for cell cycle progression but also for the structure and function of different tissues. ∞ PUBLICATIONS ∞ Hydbring P, Malumbres M, Sicinski P ( 2016 ). Non-canonical functions of cell cycle cyclins and cyclin-dependent ki- nases. Nat Rev Mol Cell Biol 17, 280-292. ∞ Malumbres M ( 2016 ). CDK4/6 Inhibi- tors resTORe Therapeutic Sensitivity in HER2(+) Breast Cancer. Cancer Cell 29, 243-244. ∞ Shao R, Liu J, Yan G, Zhang J, Han Y, Guo J, Xu Z, Yuan Z, Liu J, Malumbres M, Wan L, Wei W, Zou W ( 2016 ). Cdh1 regulates craniofacial development via APC-de- pendent ubiquitination and activation of Goosecoid. Cell Res 26, 699-712. ∞ Blas-Rus N, Bustos-Morán E, Pérez de Cas- tro I, de Cárcer G, Borroto A, Camafeita E, Jorge I, Vázquez J, Alarcón B, Malumbres M, Martín-Cófreces NB, Sánchez-Madrid F ( 2016 ). Aurora A drives early signalling and vesicle dynamics during T-cell acti- vation. Nat Commun 7, 11389. ∞ Sobecki M, Mrouj K, Camasses A, Parisis N, Nicolas E, Llères D, Gerbe F, Prieto S, Krasinska L, David A, Eguren M, Birling MC, Urbach S, Hem S, Déjardin J, Malumbres M, Jay P, Dulic V, Lafontaine DLj, Feil R, Fisher D ( 2016 ). The cell proliferation an- tigen Ki-67 organises heterochromatin. Elife 5, e13722. ∞ Quereda V, Porlan E, Cañamero M, Dubus P, Malumbres M ( 2016 ). An essential role for Ink4 and Cip/Kip cell-cycle inhibitors in preventing replicative stress. Cell Death Differ 23, 430-441. ∞ Correia NC, Melão A, Póvoa V, Sarmento L, Gómez de Cedrón M, Malumbres M, Enguita FJ, Barata JT ( 2016 ). microRNAs regulate TAL1 expression in T-cell acute lymphoblastic leukemia. Oncotarget 7, 8268-8281. ∞ Liu J, Wan L, Liu J, Yuan Z, Zhang J, Guo J, Malumbres M, Liu J, Zou W, Wei W ( 2016 ). Cdh1 inhibits WWP2-mediated ubiquitination of PTEN to suppress tum- origenesis in an APC-independent manner. Cell Discover 2, 15044. ∞ AWARDS AND RECOGNITION ∞ Elected EMBO Member. ANNUAL REPORT 2016 36 VICE-DIRECTION OF BASIC RESEARCH GENOMIC INSTABILITY GROUP Óscar Fernández-Capetillo Group Leader Staff Scientists Emilio Lecona, Matilde Murga, Sergio Ruíz SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 37 MOLECULAR ONCOLOGY PROGRAMME | GENOMIC INSTABILITY GROUP OVERVIEW Our laboratory has centred its research on trying to understand how cells respond to ‘ replicative stress ’ ( RS ), which is frequent in cancer and induced by several anticancer agents. In mammals, RS is suppressed by a signalling cascade initiated by ATR and CHK1 kinases. Throughout the years, our laboratory has developed a wide battery of cellular and animal tools for the study of RS. These tools include mice with enhanced or limited ATR-CHK1 function, cell lines in which the pathway can be activated at will, and chemical inhibitors of the ATR kinase. Our studies have revealed the impact of RS on cancer and ageing, have led to drugs that can be used to test our ideas on cancer therapy, and have also unveiled the mechanisms by which these drugs kill cancer cells. Altogether, our main aim is to understand how genome maintenance is safeguarded – particularly during replication – and to exploit this knowledge as a way to fight against cancer. “ During 2016, we have investigated which tumour types could best benefit from a treatment with ATR inhibitors, the potential mechanisms of resistance to these drugs, as well as new pathways that suppress RS.” Post-Doctoral Fellow Vanesa Lafarga Graduate Students Antonio Galarreta, Cristina Mayor, Isabel Morgado, Teresa Olbrich, Federica Schiavoni Technicians Marta E. Antón, Patricia Cozar, Alicia González ( TS )*, Sara Rodrigo, María Vega *Titulado Superior ( Advanced Degree ) ANNUAL REPORT 2016 38 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS Efficacy of ATR inhibition in two preclinical models of cancer Replicative stress ( RS ) is a widespread phenomenon in cancer cells that, when persistent, leads to DNA double strand breaks and genomic instability. Besides from the basal level of RS that occurs in every cell division, the presence of oncogenes, or many of the agents used in chemotherapy, are potent inducers of RS. In mammals, RS is sensed and suppressed through a signalling cascade that is initiated with the activation of the ATR kinase. We previously hypothesised that due to the high levels of RS in certain cancers, they could be particularly dependent on a proficient RS-response. In this regard, and in collaboration with the Experimental Therapeutics Programme, we had developed chemical inhibitors of ATR that presented some anti-tumour properties in vitro. During 2016, our work in this area was focused on the identification of tumours that are particularly sensitive to ATR inhibition, as well as on the discovery of mechanisms of resistance to these chemicals. For the first area of focus, we have shown efficacy of ATR inhibitors, as monotherapy, in 2 mouse models of Ewing Sarcoma and Acute Myeloid Leukaemia ( FIGURE 1 ). Regarding the second area of focus, we discovered – via genomewide CRISPR-Cas9 screening – that the levels of CDC25A, a key phosphatase controlling mitotic entry, are a key determinant of the sensitivity to ATR inhibitors in mouse and human cells. Figure 1 Impact of ATR inhibitors on Acute Myeloid Leukaemia. The images illustrate the efficacy of ATR inhibitors used as monotherapy in a mouse allograft model of Acute Myeloid Leukaemia, driven by an MLL- translocation ( Morgado-Palacin et al., Sci Signaling, 2016 ). Note the expansion of the tumour in control mice ( left ), which can be visualised by a colour gradient, and that is greatly reduced in mice treated with an ATR inhibitor. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 39 MOLECULAR ONCOLOGY PROGRAMME | GENOMIC INSTABILITY GROUP ∞ PUBLICATIONS ∞ Ray Chaudhuri A, Callen E, Ding X, Gogola E, Duarte AA, Lee JE, Wong N, Lafarga V, Calvo JA, Panzarino NJ, John S, Day A, Crespo AV, Shen B, Starnes LM, de Ruit- er JR, Daniel JA, Konstantinopoulos PA, Cortez D, Cantor SB, Fernandez-Capetillo O, Ge K, Jonkers J, Rottenberg S, Sharan SK, Nussenzweig A ( 2016 ). Replication fork stability confers chemoresistance in BRCA-deficient cells. Nature 535, 382-387. ∞ Murga M, Lecona E, Kamileri I, Díaz M, Lugli N, Sotiriou SK, Anton ME, Méndez J, Halazonetis TD, Fernandez-Capetillo O ( 2016 ). POLD3 Is Haploinsufficient for DNA Replication in Mice. Mol Cell 63,877-883. ∞ Ruiz S *, Mayor-Ruiz C *, Lafarga V, Murga M, Vega-Sendino M, Ortega S, Fernan- dez-Capetillo O ( 2016 ). A Genome-wide CRISPR Screen Identifies CDC25A as a Determinant of Sensitivity to ATR Inhib- itors. Mol Cell 62,307-313 ∞ Lecona E, Rodriguez-Acebes S, Specks J, Lopez-Contreras AJ, Ruppen I, Murga M, Muñoz J, Mendez J, Fernandez-Capetillo O ( 2016 ). USP7 is a SUMO deubiquitinase essential for DNA replication. Nat Struct Mol Biol 23, 270-277. ∞ Morgado-Palacin I *, Day A *, Murga M *, Lafarga V, Anton ME, Tubbs A, Chen HT, Ergan A, Anderson R, Bhandoola A, Pike KG, Barlaam B, Cadogan E, Wang X, Pierce AJ, Hubbard C, Armstrong SA, Nussenzweig A, Fernandez-Capetillo O ( 2016 ). Targeting the kinase activities of ATR and ATM exhibits antitumoral activity in mouse models of MLL-rearranged AML. Sci Signal 9, ra91. ∞ Nieto-Soler M *, Morgado-Palacin I *, Lafarga V, Lecona E, Murga M, Callen E, Azorin D, Alonso J, Lopez-Contreras AJ, Nussenzweig A, Fernandez-Capetillo O ( 2016 ). Efficacy of ATR inhibitors as single agents in Ewing sarcoma. Oncotarget 7, 58759-58767. ∞ Lecona E, Fernandez-Capetillo O ( 2016 ). A SUMO and ubiquitin code coordinates protein traffic at replication factories. Bioessays 38, 1209-1217. ∞ Fernandez-Capetillo O ( 2016 ). The ( elu- sive ) role of the SMC5/6 complex. Cell Cycle 15, 775-776. *co-first authors ∞ AWARDS AND RECOGNITION ∞ Elected Member of the European Molecu- lar Biology Organization ( EMBO ). Two new players that suppress replication stress in mammalian cells Besides ATR, several other factors participate in limiting the impact of RS in mammalian cells. Recent works have identified that POLD3, a subunit of the DNA polymerase Pold, participates in the repair of the breaks generated by RS, and also suggest that limiting its activity could be specifically deleterious for cancer cells. By developing a novel conditional knockout mouse strain we found that POLD3 deletion is lethal during embryonic development and also when depleted in adult mice. These severe defects were explained by a complete destabilisation of the POLd complex in the absence of POLD3, which abrogates DNA replication, raising serious doubts regarding the potential of POLD3 as an anticancer target. In independent work, we have been investigating how SUMO and ubiquitin participate in the coordination of DNA replication. Here, we identified USP7 as the first chromatin-associated SUMO deubiquitinase ( SDUB ) and revealed its essential role during DNA replication. Accordingly, USP7 inhibitors lead to RS and DNA damage. By deubiquitinating SUMO and/or SUMOylated proteins, our research revealed that USP7 contributes to keep a SUMO-rich and ubiquitin-poor environment at sites of DNA replication ; this is critical to maintain fork progression. Our current view is that USP7 is critical for controlling the traffic of replication factors, by supervising their recruitment or expulsion from replication factories. We propose that SUMOylation constitutes a ‘ stay ’ signal that recruits proteins near replication factors, with Ubiquitinated-SUMO being the ‘ go ’ signal that leads to their eviction ( FIGURE 2 ). s Figure 2 A SUMO and ubiquitin code regulates protein concentration around replisomes. Model derived from our recent work on the USP7 deubiquitinase ( Lecona et al., Nat Struct Mol Biol, 2016 ). We propose that, whereas SUMOylation is a signal for recruitment of proteins to the replisome, Ubiquitinated-SUMO constitutes a ‘ go ’ signal that drives their eviction. ANNUAL REPORT 2016 40 VICE-DIRECTION OF BASIC RESEARCH CHROMOSOME DYNAMICS GROUP Ana Losada Group Leader Staff Scientist Ana Cuadrado SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 41 MOLECULAR ONCOLOGY PROGRAMME | CHROMOSOME DYNAMICS GROUP OVERVIEW Our research focuses on a protein complex named cohesin that is essential for chromosome organisation. Cohesin mediates sister chromatid cohesion and, thereby, ensures faithful DNA repair by homologous recombination and proper chromosome segregation during cell division. It also plays a major role in the spatial organisation of the genome by promoting long-range DNA looping, which in turn contributes to transcriptional regulation, organisation of DNA replication factories and locus rearrangement by recombination. Mutations in cohesin have recently been found in several tumour types, most prominently in bladder cancer and acute myeloid leukaemia. Mutations in cohesin and its regulatory factors are also at the origin of a group of human syndromes collectively known as cohesinopathies. Our goal is to understand how cohesin works, how it is regulated and how its dysfunction contributes to cancer and other human diseases. In particular, we are intrigued by the existence of different versions of the cohesin complex in somatic cells. We use mouse models carrying knock out alleles of genes encoding cohesin subunits to investigate their functional specificity, both at the cellular level and in the context of an organism. We also take advantage of the Xenopus egg cell-fee system to explore additional aspects of cohesin regulation. “ We aim to define the specific contributions of cohesin-SA1 and cohesin-SA2 to genome organisation. Our work may uncover vulnerabilities in cancer cells carrying mutations in the gene encoding SA2, which is one of the twelve genes most mutated in cancer.” Graduate Students Magali de Koninck, Aleksandar Kojic, Carmen M. Morales, Miguel Ruiz, Samantha Williams Technician Miriam Rodríguez ANNUAL REPORT 2016 42 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS Cohesin-SA2 regulates transcription independently of  CTCF Cohesin consists of four core subunits, SMC1, SMC3, RAD21 and SA. There are two versions of the SA subunit in vertebrate somatic cells, SA1 and SA2. We have previously reported that cohesin-SA1 is required for telomere cohesion, while cohesin-SA2 plays a major role in centromeric cohesion. In terms of transcriptional regulation, much less is known about the potential differences between these two complexes. A study in HeLa cells reported a similar genomic distribution for cohesin-SA1 and cohesin-SA2 that largely overlapped with the distribution of the architectural protein CTCF. Our initial comparison of cohesin distribution and transcriptomes of wild type and SA1-null mouse cells suggested that cohesin-SA1 could be more important for the regulation of transcription than cohesin-SA2. First, we detected an almost double number of cohesin-binding sites genome wide in SA1 null cells, and the new sites displayed reduced overlap with CTCF and TSS ( transcription start sites ). Second, we found that in a number of genes whose expression was altered in SA1 null cells, cohesin- SA2 could not replace cohesin-SA1 efficiently since cohesin occupancy was clearly reduced in these cells ( Remeseiro, Cuadrado et al., 2012 ). When we analysed chromatin states in adult mouse brain using histone marks, CTCF and cohesin distribution, we noticed again that cohesin-SA1 complexes co-occurred with CTCF at active enhancers and promoters ( Cuadrado et al., 2015 ). More recently, using a new and better SA2 antibody, we examined the binding of cohesin- SA1 and cohesin-SA2 along the genome of human mammary epithelial cells. We found more than 20,000 positions common for both complexes in which CTCF is also present, but also found around 10,000 cohesin-SA2 specific positions, lacking SA1, in which the overlap with CTCF is significantly lower. Importantly, these cohesin-SA2 specific positions are highly enriched in enhancers and promoters and contain transcription factor binding motifs other than CTCF. Proteomic analysis of immunoprecipitates obtained with SA1 and SA2 antibodies detected several transcription factors among the SA2-specific interactors. Functional analyses, in which we compare changes in gene expression, cohesin distribution as well as chromatin architecture changes after downregulation of SA1, SA2 or CTCF, further support a prominent role of cohesin-SA2 in promoting local interactions between cis-regulatory elements independently of CTCF, while cohesin-SA1 would instead collaborate with CTCF in the demarcation of domain boundaries ( FIGURE 1 ). Pds5 proteins regulate cohesin dynamics Two factors are associated with chromatin-bound cohesin, Pds5 and Wapl. Wapl promotes cohesin unloading and in its absence there is an excess of cohesin on chromatin, and chromosome organisation is altered both in interphase and mitosis. The role of Pds5 is less clear. Moreover, there are two versions of Pds5 present in vertebrate cells : Pds5A and Pds5B. In order to explore their specific functions we previously generated murine knock out ( KO ) alleles for these two genes. We showed that both Pds5A and Pds5B contributed to cohesion establishment during S phase by promoting cohesin acetylation and Sororin binding, with Pds5B being specifically required for cohesion at Figure 1 Cohesin-SA1 and cohesin- SA2 have non-overlapping functions in genome organisation and gene regulation. Cohesin-SA2 stabilises local interactions between cis-regulatory elements ( enhancers and promoters ) mediated by transcription factors ( TF ) while cohesin-SA1 collaborates with CTCF in the demarcation of domain boundaries. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 43 MOLECULAR ONCOLOGY PROGRAMME | CHROMOSOME DYNAMICS GROUP centromeres ( Carretero et al. 2013 ). Now we have observed that cells lacking Pds5A, Pds5B, or both, have distinct alterations in their transcriptomes when compared to wild type cells. Genome wide distribution of cohesin is not obviously altered in the absence of either Pds5 protein, but does change in the absence of both. Under this condition, the dynamic association of cohesin to chromatin, measured in Fluorescence Recovery After Photobleaching ( FRAP ) experiments, is significantly decreased. Much milder effects are observed in cells lacking only Pds5A or Pds5B. Aberrant accumulation of cohesin in axial structures, known as vermicelli, previously described in Wapl depleted cells, can be observed only in the absence of the two Pds5 proteins ( FIGURE 2 ). Thus, Pds5 proteins are required for proper cohesin dynamics and cohesin distribution, although no clear specificities can be found for Pds5A and Pds5B, at least globally. The gene expression differences described above might therefore be caused by preferential interactions with chromatin regulators at specific loci. It is also possible that Pds5 proteins have functions independent of cohesin. We are currently exploring both these possibilities. s ∞ PUBLICATIONS ∞ De Koninck M, Losada A ( 2016 ). Cohesin mutations in cancer. Cold Spring Harb Perspect Med 6, a026476. ∞ Gómez R, Felipe-Medina N, Ruiz-Torres M, Berenguer I, Viera A, Pérez S, Barbero JL, Llano E, Fukuda T, Alsheimer M, Pendás AM, Losada A, Suja J ( 2016 ). Sororin loads to the synaptonemal complex central re- gion independently of meiotic cohesin complexes. EMBO Rep 17, 695-707. ∞ Abe T, Kawasumi R, Arakawa H, Hori T, Shi- rahige K, Losada A, Fukagawa T, Branzei D ( 2016 ). Chromatin determinants of the inner-centromere rely on replication fac- tors with functions that impart cohesion. Oncotarget 7, 67934-67947. ∞ Williams SJ, Abrieu A, Losada A ( 2016 ). Bub1 targeting to centromeres is suffi- cient for Sgo1 recruitment in the absence of kinetochores. Chromosoma. PMID : 27116032. Figure 2 Image of interphase nuclei from mouse embryo fibroblasts lacking both Pds5A and Pds5B ( Pds5 DKO, top ) or wild type ( WT, bottom ), fixed and stained with an antibody against cohesin SMC3 and DAPI. ANNUAL REPORT 2016 44 VICE-DIRECTION OF BASIC RESEARCH DNA REPLICATION GROUP Juan MéndezGroup Leader Post-Doctoral FellowSara Rodríguez SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 45 MOLECULAR ONCOLOGY PROGRAMME | DNA REPLICATION GROUP OVERVIEW Our laboratory studies the molecular mechanisms that underlie genomic duplication in mammalian cells. The ‘ replisome ’ complex in charge of DNA replication encounters natural obstacles ( e.g. unusual DNA structures, collisions with transcription proteins ), as well as exogenous challenges such as ionising radiation, UV light and chemicals that modify the DNA structure and block DNA polymerases. The situations in which replication forks are forced to slow down, stall or collapse are generically referred to as replicative stress ( RS ). Our Group investigates the ‘ DNA damage tolerance ’ pathways that facilitate DNA replication in the presence of RS or damaged DNA. In recent years, we have identified 2 mechanisms that counteract RS : ( 1 ) the conditional activation of dormant replication origins ; ( 2 ) the participation of PrimPol, a primase-polymerase enzyme, in the restart of stalled forks. We continue to characterise the different cellular responses to RS ( FIGURE 1 ). “ We have developed genetic tools to investigate the physiological impact of defective DNA replication, including mouse strains that suffer from a high incidence of haematological cancers due to their inefficient response to DNA damage during replication.” Graduate Students Marcos Díaz, Daniel González, Karolina Jodkowska, Sergio Muñoz ANNUAL REPORT 2016 46 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS Cellular functions of PrimPol protein We have continued to characterise PrimPol, a DNA primase- polymerase that participates in DNA damage tolerance during chromosomal replication. With this aim in mind, we have generated human and mouse cells in which PrimPol expression is either downregulated or completely ablated. PrimPol-deficient cells display a marked sensitivity to UV irradiation, including the accumulation of unrepaired Thy dimers ( CPDs and 6,4pp ) in the DNA. The skin of PrimPol KO mice also presents an inefficient healing response to UV irradiation and a higher frequency of benign papillomas. The importance of PrimPol as a tumour suppressor gene is currently being investigated. Effects of DNA re-replication in vivo CDC6 and CDT1 proteins are responsible for the loading of MCM2-7, the DNA helicase, at replication origins. After CDC6 and CDT1 execute their ‘ origin licensing ’ functions in the G1 phase, their activities are inhibited until mitosis is complete in order to prevent origin reactivation and DNA over-replication within the same cell cycle. However, these strict control mechanisms may be partially overridden in some cancer types, notably non- small cell lung carcinomas, by the overexpression of Cdc6 and/ or Cdt1 genes. We have recapitulated the deregulated expression of Cdc6 and Cdt1 using mouse strains that allow the inducible expression of both proteins, alone or in combination. While individual deregulation of CDC6 or CDT1 has only mild effects, their combination is lethal for developing embryos and also for adult individuals. Using single-molecule analysis of DNA replication, high-throughput confocal microscopy and histopathology, we have identified origin re-firing events that are sufficient to cause DNA over-replication and DNA damage in different tissues. These mouse models will allow a complete study of the physiological impact of DNA re-replication in vivo. Evidence for replicative stress in early embryonic cell cycles Replicative stress is normally studied in the context of cells undergoing external challenges. However, it also occurs in the unperturbed S phase when the replication machinery reaches special DNA structures ( e.g. G-quadruplexes ) that are difficult to replicate, or when it collides with a transcriptional fork. In 2016, we participated in a collaborative study, led by Dr M. Lopes ( University of Zurich, Switzerland ), that identified unexpected levels of physiological RS. Mouse embryonic stem cells and early embryos at the blastocyst stage display a constitutive accumulation of RPA-covered ssDNA, fork slowing and fork remodelling events, all hallmarks of RS. These characteristics are related to the short duration of the G1 phase in embryonic stem cells and are lost upon the onset of cell differentiation. This result underscores the importance of the G1 phase to fully repair DNA that had been damaged in the previous cell cycle, before entering a new round of replication ( Ahuja et al., 2016 ). Single-molecule analyses of DNA replication As replicative stress impinges on many cellular processes, the possibility of analysing DNA replication at the single-molecule level continues to attract the interest of many research groups at the CNIO and other institutions. In 2016, we collaborated in two projects led by Oscar Fernández-Capetillo ( CNIO Genomic Instability Group ) to demonstrate that USP7 ubiquitin protease targets SUMO and is essential for DNA replication ( Lecona et al., 2016 ), and that PolD3 is haploinsufficient for DNA replication in mice ( Murga et al., 2016 ). In the latter project, the analyses of replication in stretched DNA fibres revealed a striking accumulation of asymmetric forks in the absence of POLD3, a regulatory subunit of DNA polymerase δ ( FIGURE 2 ). Finally, a collaboration with R. Freire ( Hospital Universitario de Canarias, Tenerife ) revealed a novel function for USP37 ubiquitin protease in the control of DNA replication ( Hernández-Pérez et al., 2016 ). s ∞ PUBLICATIONS ∞ Murga M, Lecona E, Kamileri I, Díaz M, Lugli N, Sotiriou SK, Anton ME, Méndez J, Halazonetis TD, Fernández-Capetillo O ( 2016 ). POLD3 is haploinsufficient for DNA replication in mice. Mol Cell 63, 877-883. ∞ Lecona E, Rodríguez-Acebes S, Specks J, López-Contreras A, Ruppen I, Murga M, Muñoz J, Méndez J, Fernández-Capetillo O ( 2016 ). USP7 is a SUMO deubiquitinase essential for DNA replication. Nat Struct Mol Biol 23, 270-277. ∞ Ahuja AK, Jodkowska K, Teloni F, Bizard AH, Zellweger R, Herrador R, Ortega S, Hickson I, Altmeyer M, Méndez J, Lopes M ( 2016 ). A short G1 phase imposes con- stitutive replication stress and extensive fork remodeling in mouse embryonic stem cells. Nat Commun 7, 10660. ∞ Hernández-Pérez S, Cabrera E, Amoedo H, Rodríguez-Acebes S, Koundriouk- off S, Debatisse M, Méndez J, Freire R ( 2016 ). USP37 deubiquitinates Cdt1 and contributes to regulate DNA replication. Mol Oncol 10, 1196-1206. ∞ Muñoz S, Méndez J ( 2016 ). DNA replica- tion stress : from molecular mechanisms to human disease. Chromosoma. PMID : 26797216. ∞ Boskovic J, Bragado-Nilsson E, Prabhakar BS, Yefimenko I, Martínez-Gago J, Muñoz S, Méndez J, Montoya G ( 2016 ). Molecular architecture of the recombinant human MCM2-7 helicase in complex with nucle- otides and DNA. Cell Cycle 15, 2431-2440. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 47 MOLECULAR ONCOLOGY PROGRAMME | DNA REPLICATION GROUP Figure 1 Pathways that facilitate fork progression through damaged DNA. The yellow star represents a polymerase-blocking lesion. The main proteins involved in each pathway are mentioned. ( A ) Repriming downstream of the lesion. ( B ) Fork reversal and restart. ( C ) Translesion synthesis DNA polymerases. ( D ) Lesion skipping by template-switch. Adapted from Muñoz and Méndez ( 2016 ). Figure 2 Detection of fork asymmetry in stretched DNA fibres. Top  : representative images of replicating DNA molecules labelled with CldU ( red ) and IdU ( green ). Each image shows 2 forks moving away from a central origin. Bottom : quantification of fork asymmetry in DNA fibres prepared from B-cells from PolD3-competent ( left ) or PolD3-deficient ( right ) mice. Adapted from Murga et al ( 2016 ). ANNUAL REPORT 2016 48 VICE-DIRECTION OF BASIC RESEARCH MELANOMA GROUP María S. SoengasGroup Leader Staff ScientistDavid Olmeda SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 49 MOLECULAR ONCOLOGY PROGRAMME | MELANOMA GROUP OVERVIEW Melanomas are inherently aggressive cancers for which basic and translational research have significantly improved patient prognosis. Nevertheless, clinical responses are still incomplete. The long-term goals of our Group are to identify new progression biomarkers and therapeutic agents. Focusing on stress response programmes involving apoptosis, autophagy and endosome mobilisation, we have discovered lineage-specific oncogenes that define the melanoma ‘ fingerprint ’. Transcriptomic and proteomic analyses of the melanoma secretome have enabled us to define how tumour cells remodel the ( lymph )angiogenic vasculature and avoid immune recognition. Moreover, we have generated a unique set of animal models for non-invasive imaging of melanoma progression in vivo. These systems have led to the validation of nanoparticle-based treatments that are currently being tested in clinical trials. Our ultimate objective is to improve the management of patients with otherwise refractory metastatic melanomas. “ We have identified oncogenic cascades that are uniquely deregulated in melanoma and as such, may represent novel targets for therapeutic intervention.” Post-Doctoral Fellows Lisa Osterloh ( until January ), Paula Penacchi, Susana Frago ( since August ) Graduate Students Xavier Catena ( since June ), Daniela Cerezo, Metehan Cifdaloz, Marta Contreras ( since June ), Panagiotis Karras, Raúl Martínez, Javier Suárez ( since June ), Cristina Tejedo Technicians Tonantzin Calvo, Estela Cañón ( TS )* *Titulado Superior ( Advanced Degree ) ANNUAL REPORT 2016 50 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS CNIO Melanoma Group : objectives and model systems Melanomas are aggressive solid tumours and provide a prime example of how integrated basic and clinical research have significantly improved patient prognosis. Yet, despite great success with targeted and immune-based therapies, sustained clinical responses are still limited. Moreover, the field lacks molecular markers of diagnosis, and the knowledge of how melanomas progress and metastasise is still largely incomplete. Therefore, these questions represent key unmet needs, as emphasised by a committee of experts in which M. Soengas participates ( Merlino et al., Pigment Cell Melanoma Res 2016 ). In addition, one of the main hurdles slowing progress in this disease is the lack of animal models to monitor melanoma initiation and progression in vivo. To this end, our Group focuses on 3 main areas of research ( FIGURE 1 ): ɗ Aim 1. Oncogenic pathways, which are selectively deregulated in melanoma and may represent new diagnostic indicators. ɗ Aim 2. Risk factors and prognostic markers that underlie the unique ability of melanoma to metastasise from seemingly thin lesions. ɗ Aim 3. Animal models that allow for non-invasive monitoring of premetastatic niches, and as such, may serve as a platform for cost-effective genetic and pharmacological screens. Lineage-specific oncogenic dependencies in melanoma One of the long-term objectives of the Melanoma Group is the discovery of new melanoma drivers. We previously identified a cluster of endolysosomal-associated genes that distinguish melanoma from over 35 additional malignancies ( Alonso-Curbelo et al., Cancer Cell 2014 and Oncotarget 2015 ). In collaboration with the group of P. Agostinis ( University of Leuven, Belgium ), we further explored therapeutically-relevant regulatory mechanisms and functions of the endolysosomal machinery in different cell types ( Maes et al., FEBS J 2016 ). More recently, we also discovered unique features of autophagy ( another key lysosomal-associated process ) in melanoma. Employing human melanoma biopsies, combined with newly-generated mouse models, we identified selective heterozygous loses of ATG5 as a new risk factor for melanoma progression and as a main mediator of the resistance to targeted therapy ( García-Fernández et al. Autophagy, 2016 ). RNA binding proteins and RNA-based anticancer agents in the control of melanoma cell proliferation and metastasis Melanomas are long-known for being associated with a plethora of changes in mRNA gene expression profiles. Still, the specific contribution of RNA binding proteins ( RBPs ), particularly, spliceosome modulators, remains virtually unexplored in this disease. We have identified tumour-selective roles of RBPs Figure 1 Main objectives of the CNIO Melanoma Group aimed at identifying new progression biomarkers and validating more efficient anticancer agents. Indicated are the main experimental systems and representative publications. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 51 MOLECULAR ONCOLOGY PROGRAMME | MELANOMA GROUP CPEB4 and CUGBP1 in the regulation of mRNA stability, with unexpected targets involving master specifiers of the melanocyte lineage ( FIGURE 2 ). We have also assisted F. Gebauer’s laboratory ( the Centre for Genomic Regulation, Barcelona ) with the identification of pro-metastatic roles of the translation controller UNR in melanoma ( Wuth et al., Cancer Cell 2016 ). Similarly, histopathological studies with our long-term collaborators, P. Ortiz-Romero and J.L. Rodríguez-Peralto ( Hospital 12 de Octubre, Madrid ), have validated the chromatin remodeler and RNA binding factor DEK as a risk factor for melanoma metastasis ( Riveiro-Falkenbach, Pigment Cell Melanoma Res, 2016 ). We have also made great progress regarding one of the most pressing needs in the field of melanoma, namely, the mechanisms underlying immune suppression ( reviewed in Cerezo-Wallis and Soengas, Curr Pharm Design 2016 ). This was achieved by combining the analysis of human melanoma biopsies with a new class of ‘ Lymphoreporter ’ mouse models that we generated in collaboration with Sagrario Ortega’s Transgenic Mice Unit at the CNIO. Moreover, we have expanded the use of dsRNA nanoparticles as immunomodulatory agents. This information will be used to support clinical trials that are currently being performed by Bioncotech Therapeutics, a biotechnology company cofounded by M. Soengas. s ∞ PUBLICATIONS ∞ Wurth L, Papasaikas P, Olmeda D, Bley N, Calvo GT, Guerrero S, Cerezo-Wallis D, Martinez-Useros J, García-Fernández M, Hüttelmaier S, Soengas MS, Gebauer F ( 2016 ). UNR/CSDE1 Drives a Post-tran- scriptional Program to Promote Melanoma Invasion and Metastasis. Cancer Cell 30, 694-707. ∞ Casas C, Codogno P, Pinti M, Batoko H, Morán M, Proikas-Cezanne T, Reggiori F, Sirko A, Soengas MS, Velasco G, Lafont F, Lane J, Faure M, Cossarizza A ( 2016 ). TRANSAUTOPHAGY : European network for multidisciplinary research and transla- tion of autophagy knowledge. Autophagy 12, 614-617. ∞ García-Fernández M, Karras P, Checinska A, Cañón E, Calvo GT, Gómez-López G, Cifdaloz M, Colmenar A, Espinosa-Hevia L, Olmeda D, Soengas MS ( 2016 ). Metastatic risk and resistance to BRAF inhibitors in melanoma defined by selective allelic loss of ATG5. Autophagy 12, 1776-1790. ∞ Pérez-Guijarro E, Karras P, Cifdaloz M, Martínez-Herranz R, Cañón E, Graña O, Horcajada-Reales C, Alonso-Curbelo D, Calvo TG, Gómez-López G, Bellora N, Riveiro-Falkenbach E, Ortiz-Romero PL, Rodríguez-Peralto JL, Maestre L, Ronca- dor G, de Agustín Asensio JC, Goding CR, Eyras E, Megías D, Méndez R, Soengas MS ( 2016 ). Lineage-specific roles of the cyto- plasmic polyadenylation factor CPEB4 in the regulation of melanoma drivers. Nat Commun 7, 13418. ∞ Martínez-Iglesias O, Olmeda D, Alonso-Me- rino E, Gómez-Rey S, González-López AM, Luengo E, Soengas MS, Palacios J, Regad- era J, Aranda A ( 2016 ). The nuclear core- pressor 1 and the thyroid hormone receptor ß suppress breast tumor lymphangiogen- esis. Oncotarget 7, 78971-78984. ∞ Merlino G, Herlyn M, Fisher DE, Bastian BC, Flaherty KT, Davies MA, Wargo JA, Curiel-Lewandrowski C, Weber MJ, Leachman SA, Soengas MS, McMahon M, Harbour JW, Swetter SM, Aplin AE, Atkins MB, Bosenberg MW, Dummer R, Gershenwald JE, Halpern AC, Herlyn D, Karakousis GC, Kirkwood JM, Krautham- mer M, Lo RS, Long GV, McArthur G, Ribas A, Schuchter L, Sosman JA, Smalley KS, Steeg P, Thomas NE, Tsao H, Tueting T, Weeraratna A, Xu G, Lomax R, Martin A, Silverstein S, Turnham T, Ronai ZA ( 2016 ). The state of melanoma : challenges and opportunities. Pigment Cell Melanoma Res 29, 404-416. ∞ Riveiro-Falkenbach E, Ruano Y, García- Martín RM, Lora D, Cifdaloz M, Acquadro F, Ballestin C, Ortiz-Romero PL, Soengas MS, Rodríguez-Peralto JL ( 2016 ). DEK onco- gene is overexpressed during melanoma progression. Pigment Cell Melanoma Res. PMID : 27893188. ∞ García-Pascual C et al. (incl. Soengas MS) (2016). Evaluation of the antiproliferative, proapoptotic, and antiangiogenic effects of a double-stranded RNA mimic com- plexed with polycations in an experimental mouse model of leiomyoma. Fertil Steril 105, 529-538. ∞ Maes H, Olmeda D, Soengas MS, Agostinis P ( 2016 ). Vesicular trafficking mechanisms in endothelial cells as modulators of the tumor vasculature and targets of antian- giogenic therapies. FEBS J 283, 25-38. ∞ Cerezo-Wallis D, Soengas M ( 2016 ). Un- derstanding Tumor-Antigen Presentation in the New Era of Cancer Immunotherapy. Curr Pharm Design. PMID : 27568731. ∞ AWARDS AND RECOGNITION ∞ Outstanding Research Award, the Society for Melanoma Research. ∞ Elected Board Member of ASEICA (Aso- ciación Española de Investigación Sobre el Cáncer). Figure 2 New melanoma drivers : the RNA binding protein CPEB4. ( A ) Upregulation of CPEB4 in malignant melanomas visualised by comparative histological analyses in benign nevi and malignant tumour specimens. CPEB4 is stained in pink. (  B ) Schematic representation of newly identified roles of CPEB4 in the control of lineage-specific tumour drivers ( MITF and RAB27 ). These functions link, for the first time, RNA binding proteins to mechanisms underlying tumour type identity. ANNUAL REPORT 2016 52 VICE-DIRECTION OF BASIC RESEARCH MICROENVIRONMENT & METASTASIS JUNIOR GROUP OVERVIEW Our laboratory is focused on understanding metastatic progression. During this process, tumour cells communicate actively with the tumour microenvironment. Among all factors involved in metastasis, our laboratory is specifically interested in defining the role of secreted exosomes during pre-metastatic niche formation. Exosomes are actively involved in cell-cell communication during both physiological and pathological processes. Our data support that tumour-secreted exosomes are involved in : 1 ) pre-metastatic niche formation and metastatic organotropism depending on the integrin expression profile on their surface ; and 2 ) stromal cell reprogramming by horizontal transfer of molecules ( i.e. oncoprotein c-MET ) and/or influencing the expression of pro-inflammatory and pro-vasculogenic molecules. “ Exosome secretion by metastatic cells is an adaptive strategy for tumour cells to corrupt the surrounding microenvironment, thereby favouring tumour progression.” Héctor Peinado Junior Group Leader Staff Scientist Susana García Post-Doctoral Fellows Marta Hergueta, Claudia Savini ( since November ) Graduate Students Ana I. Amor, Teresa González ( since June ), Lucía Robado SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 53 MOLECULAR ONCOLOGY PROGRAMME | MICROENVIRONMENT & METASTASIS JUNIOR GROUP RESEARCH HIGHLIGHTS Role of tumour-derived exosomes in lymph node metastasis Melanoma-secreted exosomes have been shown to home to specific niches in lymph nodes. We are studying how tumour- secreted exosomes promote cellular and molecular alterations in the lymph node microenvironment, fostering metastasis ( FIGURE, A ). The goal of the current project is to determine the mechanisms through which tumour-derived exosomes promote lymph node and distal metastasis. Our studies in melanoma patients will be the first ones evaluating the use of circulating vesicles in lymphatic fluid as biomarkers to predict relapse and metastatic potential. Linking obesity with metastatic risk Obesity has been associated with the increased risk of developing metastasis in certain cancers. Although the implication of obesity in cancer is clear, there is, to date, a lack of studies analysing the impact of obesity on metastasis. We are investigating the mechanisms involved in the crosstalk between the adipose tissue, platelets and tumour cells during the metastatic process ( FIGURE, B ). We are dissecting the systemic effects of tumour- derived exosomes in adipose tissue as well as the involvement of platelets, determining their role in metastasis. Ultimately, we aim to determine specific signatures in circulating exosomes and platelets of cancer patients in order to define new prognostic and therapeutic markers that can be applied in the clinical setting. Novel pathways involved in neurofibromatosis progression Although neurofibromatosis is a genetic disorder, in this project we aim to develop a very innovative concept, which focuses on unveiling unknown pathways involved in exosome secretion during neurofibromatosis progression. We are investigating the molecular signature of exosomes secreted from highly metastatic neurofibromatosis models. Our data support that tumour-secreted exosomes carry a specific signature that can be detected in the circulation. This approach will result in the development of new diagnostic tests and therapies to block neurofibromatosis progression. s Technicians Marina Mazariegos, Cristina Merino, Sara Sánchez-Redondo ( since June ) Visiting Scientist Olwen Leaman ∞ PUBLICATIONS ∞ Becker A, Thakur BK, Weiss JM, Kim HS, Peinado H, Lyden D ( 2016 ). Extracellular Vesicles in Cancer : Cell-to-Cell Mediators of Metastasis. Cancer Cell 30, 836-848. ∞ García-Silva S, Peinado H ( 2016 ). Melano- somes foster a tumour niche by activating CAFs. Nat Cell Biol 18, 911-913. ∞ Zhou J, Ghoroghi S, Benito-Martin A, Wu H, Unachukwu UJ, Einbond LS, Guariglia S, Peinado H, Redenti S ( 2016 ). Charac- terization of Induced Pluripotent Stem Cell Microvesicle Genesis, Morphology and Pluripotent Content. Sci Rep 6, 19743. ∞ Torrano V, Royo F, Peinado H, Loizaga-Iri- arte A, Unda M, Falcón-Perez JM, Carrace- do A ( 2016 ).Vesicle-MaNiA : extracellular vesicles in liquid biopsy and cancer. Curr Opin Pharmacol 29, 47-53. ∞ Veerappan A, Thompson M, Savage AR, Silverman ML, Chan WS, Sung B, Summers B, Montelione KC, Benedict P, Groh B, Vicencio AG, Peinado H, Worgall S, Silver RB ( 2016 ). Mast cells and exosomes in hyperoxia-induced neonatal lung disease. Am J Physiol Lung Cell Mol Physiol 310, L1218-L1232. ∞ AWARDS AND RECOGNITION ∞ XVII Fundacion Pfizer Research Award 2016, Spain. ∞ FERO Grant for Translational Research in Oncology ( XI BECA FERO 2016 ), FERO Foundation for Oncology Research, Spain. Figure ( A ) Analysis of exosome distribution in sentinel lymph nodes. Green-labelled exosomes from B16- F1R2 melanoma cells were injected in the footpad and followed for 16 hours. Analysis of lymph nodes demonstrated that exosomes reach popliteal ( sentinel ) lymph nodes with a specific distribution found mainly in subcortical areas co-localising with lymphatic endothelial cells ( in red ). ( B ) Metastasis of breast cancer cell lines in lung metastatic niches. Tumour breast cancer cell lines ( in red ) were injected by tail vein in mice, in combination with platelets. Analysis of metastasis demonstrates that tumour cells reach metastatic lungs in areas surrounding terminal bronchioles, formerly known as areas where pre-metastatic niches were formed. B A ANNUAL REPORT 2016 54 VICE-DIRECTION OF BASIC RESEARCH BRAIN METASTASIS JUNIOR GROUP OVERVIEW Brain metastasis is the most common neurological complication of cancer. When metastatic cells reach the brain, prognosis is poor given that available therapies ( i.e. surgery and radiation ) have limited benefits for patients and the disease inevitably progresses. The rise in the number of patients with brain metastasis is partially due to the increasing number of systemic therapies that work extracranially but not in the brain. In this scenario, cancer cells present at this highly demanding secondary site have additional time to evolve and develop into clinically detectable lesions. In the laboratory, we study why and how cells from different cancer types ( breast cancer, lung cancer and melanoma ) are able to access the brain, survive and colonise this vital organ. We dissect the biology of these processes in vivo using experimental models in order to challenge the current status of this unmet clinical need. “ The Brain Metastasis Group is seeking to identify novel ways to target both cancer cells and the associated microenvironment in order to reduce metastatic burden in the brain.” Manuel Valiente Junior Group Leader Post-doctoral Fellow Neibla Priego ( since April ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 55 MOLECULAR ONCOLOGY PROGRAMME | BRAIN METASTASIS JUNIOR GROUP RESEARCH HIGHLIGHTS The Brain Metastasis Group investigates the progression of cancer to the Central Nervous System ( CNS ). During 2016, we focused our efforts on various projects : ɗ Using a novel medium-throughput drug discovery platform, the laboratory identified two compounds with the potential to target established brain metastasis from experimental lung and breast cancer models. ɗ We identified two novel mediators of brain metastasis that are enabling us to explore the influence of epigenetics on brain colonisation as well as the ability of cancer cells to interact with neurotransmitters. ɗ We are evaluating the therapeutic potential of targeting specific components of the microenvironment that are only present surrounding metastatic lesions in the brain. Our research suggests that the viability of brain metastasis is highly dependent on altered components of the microenvironment, thus highlighting potential vulnerabilities. s Graduate Students Catia P. Domingues, Maria Figueres ( until February ), Almudena Saiz ( until October ), Lucia Zhu ( since September ) Technician Laura E. Doglio ( since January ) Students in Practice Manon Mulders ( until June ), David Wasilewski ( until May ), Pablo Sánchez ( July-September ), Marta Serrano ( June-August ), Zira Dorado ( January-July ), Carmen Díaz ( May- June ) ∞ PUBLICATIONS AT OTHER INSTITUTIONS ∞ Chen Q *, Boire A *, Jin X, Valiente M, Er EE, Lopez-Soto A, Patwa R, Sha H, Xu K, Cross JR, Massagué J ( 2016 ). Carcinoma-astrocyte gap junctions promote brain metastasis by cGAMP transfer. Nature 533, 493-498. *Shared authorship. ∞ AWARDS AND RECOGNITION ∞ IV “ Profesor Durántez ”-Fundación LAIR Award 2016, Spain. Figure The biology of brain metastasis is at the centre of all our projects. We are developing better pre-clinical models, discovering novel mediators required for the colonisation of the brain and are dissecting molecular interactions with the microenvironment in order to explore new therapies. ANNUAL REPORT 2016 56 VICE-DIRECTION OF BASIC RESEARCH METABOLISM AND CELL SIGNALLING JUNIOR GROUP OVERVIEW In the Metabolism and Cell Signalling Lab, we study the interplay between nutrients, metabolism and cancer. The alarming increase of overweight and obesity over the last decades and the epidemiological links between elevated nutrient levels and human disease calls for a better understanding of the molecular underpinnings of these connections. Conversely, limiting nutrient intake to an extent that does not cause malnutrition is not only protective against diabetes, but also prevents cancer development and delays ageing in most multicellular species by mechanisms that are poorly understood. In the lab, we combine mouse genetics and cell biological tools to gain insight into the cellular processes that become corrupted upon nutrient overabundance, aiming to conceive therapeutic interventions targeting these processes in the context of cancer and the process of ageing. “ By means of novel strains of mice with deregulated nutrient sensing we identified metabolic alterations that drive uncontrolled proliferation of B lymphocytes and nutrient imbalances associated with diabetes.” Alejo Efeyan Junior Group Leader Post-Doctoral Fellow Ana Ortega SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 57 MOLECULAR ONCOLOGY PROGRAMME | METABOLISM AND CELL SIGNALLING JUNIOR GROUP RESEARCH HIGHLIGHTS Mammals, including humans, have evolved in an environment where the ability to efficiently use limiting nutrient sources has been a key survival adaptation that has shaped all our responses to nutrients. Unprecedented nutrient overabundance is in conflict with our cellular and organismal responses, which are best tuned to operate under scarcity. These aberrant responses not only lie at the core of the pathogenesis of the metabolic alterations observed in diabetes, but are also key in cancer and the process of ageing. We use genetically engineered strains of mice as a physiological framework to understand the molecular bridges from elevated nutrient levels to human disease. In particular, we have genetically modified the RagA and RagC GTPases, key players in the sensing of nutrients that activate a master regulator of metabolism, a kinase called mTOR. Mice with gain-of-function mutations in RagA − therefore unable to sense a drop in nutrient levels − have an increased glycaemia in spite of a normal food intake and decreased adiposity. Furthermore, these mice show intolerance to glucose, which means that when glucose is administered it remains in the circulation, and peripheral organs ( such as liver and skeletal muscle ) are unable to uptake it. These perturbations are tightly associated with the development of type 2 diabetes. Indeed, when we examined the ability of peripheral tissues to respond to insulin we observed an impaired response to insulin, also known as insulin resistance, which leads to increased levels of glucose in circulation. We are currently characterising other metabolic imbalances observed in these mice and are performing a genetic dissection of these alterations by deregulating nutrient sensing in an organ-specific manner. s Graduate Students Celia de la Calle, Nerea Deleyto Technician Ana Sagrera ( since August ) ( TS )* *Titulado Superior ( Advanced Degree ) Student in Practice Andrew Vandenberg ( until August ) ∞ PUBLICATIONS AT OTHER INSTITUTIONS ∞ Okosun J, Wolfson RL, Wang J, Araf S, Wilkins L, Castellano BM, Escudero-Ibarz L, Al Seraihi AF, Richter J, Bernhart SH, Efeyan A, Iqbal S, Matthews J, Clear A, Guerra-As- sunção JA, Bödör C, Quentmeier H, Mansbridge C, Johnson P, Davies A, Strefford JC, Packham G, Barrans S, Jack A, Du MQ, Calaminici M, Lister TA, Auer R, Montoto S, Gribben JG, Siebert R, Chelala C, Zoncu R, Sabatini DM, Fitzgibbon J ( 2016 ). Recurrent mTORC1-activating RRAGC mutations in follicular lymphoma. Nat Genet 48, 183-188. Figure Mice that express mutant variants of the Rag GTPases are genetically unable to respond to a drop in nutrient levels. These mice show metabolic defects that impact on glucose homeostasis and on the control of B lymphocyte behaviour. ANNUAL REPORT 2016 58 VICE-DIRECTION OF BASIC RESEARCH CANCER CELL BIOLOGY PROGRAMME ERWIN F. WAGNER Programme Director SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 59 CANCER CELL BIOLOGY PROGRAMME The overall strategic goals of the Cancer Cell Biology Programme are to achieve a better understanding of the events leading to cancer development, progression and metastasis, and to discover molecular mechanisms that could provide a basis for novel therapies. The 5 Groups investigate how tumours grow as ‘ extrinsic organs ’; the spectrum of investigations ranges from epithelial cancers such as liver, skin and intestinal cancer, to bone and brain tumours. The research covers aspects of tumour cell biology, ranging from tumour stem cells, tumour cell interactions with host cells/environment such as tumour-associated cells ( like macrophages and fibroblasts ), to the role of inflammation, as well as cell adhesion, metabolism and metastasis. Powerful state-of-the-art mouse genetic models, human cellular systems, high-throughput genomic/proteomic and biochemical tools, as well as patient-derived materials, are employed. At present, these aspects are successfully covered and integrated in an interactive and collaborative manner by the complementary research areas of 2 Senior and 3 Junior Groups. The Senior Group, led by Francisco X. Real, studies epithelial tumours and focuses mainly on pancreatic and bladder cancer. The Group employs an integrative approach to understand the molecular patho-physiology of these tumours and applies this knowledge in the clinical setting. Mirna Pérez-Moreno’s Group investigates the role of cell adhesion, inflammation and cellular signalling in normal skin physiology and cancer. Nabil Djouder’s Group aims to dissect the contribution of nutrient and growth factor signalling pathways to cancer development, and in particular to gastro-intestinal cancers. Massimo Squatrito’s Group, which is partly supported by the Seve Ballesteros Foundation, studies how brain tumours, mainly glioblastomas and medulloblastomas, develop and how they respond to therapy. Finally, my own Group focuses on understanding the role of the transcription factor complex AP-1 ( Fos/Jun ) in physiological and pathological processes, with a strong focus on aspects of inflammation and cancer, e.g. in the liver, lung, skin and bone. We are investigating the role of AP-1 in inflammatory skin diseases, such as psoriasis, but also aim to molecularly define the causes of lung fibrosis. We have continued our efforts to study how the whole organism responds to a locally growing tumour in the context of a complex metabolic impairment in cancer-associated-cachexia. “ Our main goal is to keep CNIO globally competitive and to ensure that CNIO remains an international institution. Fourteen different nationalities are represented in our Programme and the goal is to perform first-class cancer cell biology, as well as to train students and postdocs to become the next- generation of promising scientists.” ANNUAL REPORT 2016 60 VICE-DIRECTION OF BASIC RESEARCH GENES, DEVELOPMENT AND DISEASE GROUP Erwin F. Wagner Group Leader Staff Scientists Latifa Bakiri, Nuria Gago, María Jiménez, Liliana Fajardo Mellor, Özge Uluçkan SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 61 CANCER CELL BIOLOGY PROGRAMME | GENES, DEVELOPMENT AND DISEASE GROUP OVERVIEW Our studies aim to analyse gene function in healthy and pathological conditions, e.g. in tumour development, using the mouse as a model organism but also employing patient- derived samples. Specifically, the functions of the AP-1 ( Fos/ Jun ) transcription factor complex regulating cell proliferation, differentiation and oncogenesis, as well as the cross-talk between organs, are being investigated. The goal is to define molecular pathways that lead to disease/cancer development and to identify novel therapeutic targets ( FIGURE ). We focus on : ɗ Elucidating a causal link between inflammation, cancer and AP-1 ( Fos/Jun ) expression, using cell type-specific, switchable genetically engineered mouse models ( GEMMs ). ɗ Developing and characterising new GEMMs for cancer and human diseases, such as bone loss, fibrosis and psoriasis, and applying these to preclinical studies. ɗ Using multiple approaches to compare mouse models of disease to human disease and to identify therapeutically relevant targets. “ Our goal is for CNIO to remain an international and competitive institution. At present, 4 out of 5 Group Leaders in our department are foreigners, one of whom is partly funded by the Seve Ballesteros Foundation. Fourteen different nationalities from 4 continents are a testament to our international science culture and we all focus on unravelling the mysteries of inflammation, metabolism and cancer.” Post-Doctoral Fellows Albanderi Alfraidi, Kazuhiko Matsuoka, Álvaro Ucero Graduate Student Lucía T. Díez Technicians Vanessa Bermeo Ana Guío ( TS )* *Titulado Superior ( Advanced Degree ) ANNUAL REPORT 2016 62 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS We have developed a powerful technology for switchable, reversible and tissue-specific ectopic gene expression of specific AP-1 monomers/dimers in the liver, lung, skin and bone. We use mouse and human tissue samples for large-scale studies, such as deep sequencing ( RNA-Seq, ChIP-Seq ) and mass spectrometry analyses. We evaluate possible biomarkers and therapeutic approaches in small-scale pre-clinical studies based on these screenings. Bone development, osteosarcomas and arthritis We are studying the function of AP-1 proteins and their targets in bone development and disease using loss-( LOF ) and gain- of-function ( GOF ) mouse models. In mice, c-Fos expression leads to osteosarcomas ( OSs ) and chondrogenic hyperplasias. We found that loss of Wnt signalling delays OS development in c-Fos GOF mice, pointing to a novel mechanism linking c-Fos/ AP-1 and OS development. Rheumatoid Arthritis ( RA ), Psoriatic Arthritis ( PsA ) and Osteoarthritis ( OA ) are destructive joint pathologies linked to chronic inflammatory diseases. We are studying the function of AP-1 factors and their target genes in the development of arthritis using GEMMs, experimental arthritis models and local gene manipulation approaches. Additionally, we are investigating how crosstalk from other organs, like skin and bone, may contribute to the development and progression of different types of arthritis, as well as whether inflammation generated from the joint, using arthritic models, can influence or induce disease development in adjacent and distant organs. Liver disease – metabolism, fibrosis, inflammation and cancer AP-1 proteins are important modulators of hepatic lipid metabolism as specific AP-1 dimers can either activate or repress PPARγ transcription. Therefore, fatty liver disease and obesity most likely depend on AP-1 dimer composition. In addition, while Fra proteins protect against steatosis, ectopic expression of Fra-2, but not Fra-1-containing AP-1 dimers in hepatocytes, leads to liver dysplasia in aged mice. Mechanistically, molecular analyses point to the involvement of pathways connected to human hepatocellular carcinoma ( HCC ), such as the Wnt/β- catenin and Myc pathways. Deletion of c-Fos in hepatocytes protects from chemically- induced liver carcinogenesis, whereas additional inactivation in immune cells abrogates this protective effect. Ectopic c-Fos or expression of Fos-dimers leads to altered cholesterol and bile Figure Tet-switchable AP-1 transgenic mice were generated for ectopic expression of specific AP-1 monomers/ dimers in skin, bone, liver and lung, which are complemented by loss-of- function mouse models. Proteomics, expression profiling, RNA-sequencing and ChIP-sequencing are employed to compare mouse models of disease to human disease as well as to identify novel targets. Furthermore, we are investigating the systemic response of the mouse organism to a growing tumour in cancer cachexia. Preclinical studies are performed using different genetical ly engineered mouse models with compounds that target the identified molecules in order to determine the potential of translating our findings for the treatment of human disease. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 63 CANCER CELL BIOLOGY PROGRAMME | GENES, DEVELOPMENT AND DISEASE GROUP acid metabolism, inflammation, fibrosis, hepatocyte/bile duct proliferation and tumours with human HCC gene signatures. A robust connection between c-Fos expression and the activity of the LXR/RXR pathway, an important regulator of cholesterol homeostasis, was observed and it most likely contributes to the oncogenic function of c-Fos in hepatocytes. Cancer-associated cachexia ( CAC ) We previously demonstrated that ‘ browning ’, a switch from white to brown fat, is a contributor to the wasting process in CAC, and we also documented the importance of IL-6 and β-adrenergic signalling. Using GEMMs, as well as syngeneic mouse models, we are investigating the role of inflammation in CAC and are also studying the systemic events in CAC, such as the role of the neuro-endocrine system, e.g. the renin-angiotensin-aldosterone system. In collaboration with the Medical University of Vienna and Attoquant Diagnostics ( Vienna ), we are analysing human serum samples from cancer patients to validate the findings from the GEMMs. Our goal is to understand the molecular switch from a local inflammation-associated tumour to the systemic effects of CAC, and to potentially identify novel biomarkers ( in collaboration with Drs R. Senaris, Santiago de Compostella, Spain and M. Petruzzelli, Cambridge, UK ). Defining a function for AP-1 in lung disease Lung fibrotic diseases and non-small cell lung cancer ( NSCLC ) share the same target organ as well as similar characteristics such as higher incidence in smokers, high morbidity and lack of effective treatments leading to high mortality. Our studies using GEMMs provide experimental tools for studying the important contribution of Fra proteins to these diseases. While Fra-2 is required for the innate immune response associated with disease progression in experimental lung fibrosis, Fra-2 promotes tumour growth in NSCLC. We are currently testing the therapeutic value of Fra-2 inhibition in our pre-clinical models for fibrosis and NSCLC, and are validating our findings using patients ’ tissue samples. These studies are conducted in collaboration with Daiichi Sankyo Company ( Japan ) and Mariano Barbacid’s Experimental Oncology Group at CNIO, respectively. Skin cancer, inflammation and human disease We have demonstrated that loss of epidermal Fra-2 protein results in skin barrier defects. Mechanistically, Fra-2 binds and transcriptionally regulates epidermal differentiation gene promoters. We are currently investigating the targets of Fra-2 in skin that play a role in barrier function and inflammation. Characterisation of the epidermal inflammatory disease in mice lacking JunB suggests a skin to bone crosstalk. We have recently reported that IL-17A production in skin causes bone loss by inhibiting Wnt signalling in bone-forming osteoblasts. We have extended our studies and shown that psoriasis patients suffer from bone loss that correlates with IL-17A levels. We are currently evaluating the role of the microbiota in skin inflammation by antibiotic treatments, high-throughput microbiota sequencing and germ-free housing conditions. High-throughput proteomics and transcriptomic analyses unravelled novel pathways and molecules for targeted therapies, such as S100A8/A9 and complement C3. We have now generated new GEMMs to define the role of these novel target molecules in inflammatory skin disease with a focus on the systemic effects beyond the skin in arthritis and bone loss. Another angle of research in psoriasis involves the analysis of the role of epidermal stem cells in the disease initiation and progression using state-of-the-art lineage-tracking models. Recent data suggest that a subtype of epidermal stem cells is important for disease progression ; we are currently characterising these cells and expanding these studies to patient samples. Finally, we are using GEMMs for Squamous Cell Carcinomas ( SCCs ), previously generated in the lab, for strategies to prevent skin cancers and to develop novel therapeutic approaches to treat peri-neural invasion with reduced incidence of metastasis. s ∞ PUBLICATIONS ∞ Wagner EF ( 2016 ). Cancer : Fibroblasts for all seasons. Nature 530, 42-43. ∞ Nowell CS, Odermatt PD, Azzolin L, Hohnel S, Wagner EF, Fantner GE, Lutolf MP, Barran- don Y, Piccolo S, Radtke F ( 2016 ). Chronic inflammation imposes aberrant cell fate in regenerating epithelia through mech- anotransduction. Nat Cell Biol 18, 168-180. ∞ Uluçkan Ö, Jimenez M, Karbach S, Jeschke A, Graña O, Keller J, Busse B, Croxford AL, Finzel S, Koenders M, van den Berg W, Schinke T, Amling M, Waisman A, Schett G, Wagner EF ( 2016 ). Chronic skin inflamma- tion leads to bone loss by IL-17-mediated inhibition of Wnt signaling in osteoblasts. Sci Transl Med 8, 330ra37. ∞ Petruzzelli M, Wagner EF ( 2016 ). Mech- anisms of metabolic dysfunction in can- cer-associated cachexia. Genes Dev 30, 489-501. ∞ Blanpain C, Wagner EF ( 2016 ). Editorial Overview : The ins and outs of stem cells in differentiation, inflammation & disease. Curr Opin Cell Biol 43, iv-vi. ∞ Trierweiler C, Hockenjos B, Zatloukal K, Thimme R, Blum HE, Wagner EF, Hassel- blatt P ( 2016 ). The transcription factor c-JUN/AP-1 promotes HBV-related liver tumorigenesis in mice. Cell Death Differ 23, 576-582. ∞ Puujalka E, Heinz M, Hoesel B, Friedl P, Schweighofer B, Wenzina J, Pirker C, Schmid JA, Loewe R, Wagner EF, Berg- er W, Petzelbauer P ( 2016 ). Opposing Roles of JNK and p38 in Lymphangio- genesis in Melanoma. J Invest Dermatol 136, 967-977. ∞ Shatirishvili M, Burk AS, Franz CM, Pace G, Kastilan T, Breuhahn K, Hinterseer E, Dierich A, Bakiri L, Wagner EF, Ponta H, Hartmann TN, Tanaka M, Orian-Rousseau ( 2016 ). Epidermal-specific deletion of CD44 reveals a function in keratinocytes in response to mechanical stress. Cell Death Dis 7, e2461. ∞ Uluçkan Ö, Wagner EF. ( 2016 ). Role of IL- 17A signalling in psoriasis and associated bone loss. Clin Exp Rheumatol 34, 17-20. ANNUAL REPORT 2016 64 VICE-DIRECTION OF BASIC RESEARCH EPITHELIAL CARCINOGENESIS GROUP Francisco X. Real Group Leader Staff Scientist Victor J. Sánchez-Arevalo SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 65 CANCER CELL BIOLOGY PROGRAMME | EPITHELIAL CARCINOGENESIS GROUP OVERVIEW We focus on the molecular pathophysiology of pancreatic ductal adenocarcinoma ( PDAC ) and urothelial carcinoma ( UC ), adopting a disease-oriented approach. We use cultured cells, genetically modified mice, and patient samples, giving similar weight to the 3 model systems. Primary observations are made at either of these levels and are then extended through additional work. To translate the findings, we bring this knowledge to a ‘ population ’ level, harnessing information and samples from large patient cohorts. In PDAC, we study cell differentiation as a potent tumour suppressor mechanism acting early during carcinogenesis. We use the excellent genetic mouse models available because these processes cannot be readily studied using human samples. PDAC can originate both in pancreatic progenitors and in acinar cells. Understanding the contribution of these cell types to PDAC is crucial to design better strategies for early tumour detection and prevention in subjects at risk. “ In UC, we focus on identifying new genes, using them for improved tumour taxonomy, characterising their mechanisms of action, and applying this knowledge for improved prediction of outcome and therapy.” Post-Doctoral Fellows Enrique Carrillo, Eleonora Lapi, Miriam Marqués Graduate Students Isidoro Cobo, Francesc Madriles ( until March ), Catarina Pereira, Mónica Pérez ( since September ) Technicians Itxaso Bellón ( TS )*, Natalia Del Pozo, María Tania Lobato, Laia Richart ( until September ) ( TS )* *Titulado Superior ( Advanced Degree ) Visiting Scientist Juan R. Tejedor ( Universitat Pompeu Fabra, Barcelona ) ANNUAL REPORT 2016 66 VICE-DIRECTION OF BASIC RESEARCH RESEARCH HIGHLIGHTS Pancreas cancer molecular pathophysiology We are analysing the tumour suppressive role of several transcription factors involved in pancreatic differentiation. PDAC is characterised by highly prevalent alterations in KRAS, p16, TP53, and SMAD4, and by low-frequency alterations in a plethora of other genes converging in a few critical genetic pathways. The currently accepted progression model proposes that the sequential acquisition of these genetic changes drives the development of PanIN-1, -2, and -3 lesions. We have previously highlighted the weaknesses of this model. Using mutant KRasG 12V as the driving oncogene in a pancreatic Gata4-null background, we have shown that PDAC can be initiated from pancreatic progenitors or adult acinar cells without the development of acino-ductal metaplasia ( ADM ) or preneoplastic PanINs ( FIGURE 1 ). These findings, together with recent evidence using whole genome sequencing of human PDAC, suggest the existence of alternative mechanisms of tumour evolution and call for the identification of relevant precursor lesions. RNA-Seq analyses of KRasG 12V ;Gata4-null vs. KRasG 12V ;Gata4-wild type pancreata point to differential activation of inflammatory pathways, possibly involved in the PanIN-less phenotype. These findings converge with our data showing the requirement of a full dose of Nr5a2 to suppress inflammation in mouse and human pancreas : reduced Nr5a2 dosage leads to the activation of AP-1 and an epithelial cell-autonomous pre-inflammatory state characterised by production of chemotactic factors. Importantly, Nr5a2 controls both epithelial differentiation and inflammatory programmes, providing mechanistic evidence that these processes are linked at the transcriptional level in pancreatic cells. We are also exploring the potential of enhancing Nr5a2 activity to suppress pancreatitis and tumour development. This work benefits from a close collaboration with the CNIO Groups of E. Wagner and N. Malats. Urothelial carcinoma ( UC ) genetics, biology, and clinical translation Our goal is to refine current knowledge on the genomic landscape of UC and apply this in the clinical setting. Through exome sequencing we identified STAG2 and RBM10 as new UC genes that are more broadly involved in human cancer. STAG2 codes for a cohesin subunit ; its inactivation in UC is not associated with aneuploidy, suggesting that regulation of chromatin architecture and gene expression mediate its tumour suppressor role. Transcriptomic analyses of human tumours and cultured cells, as well as biochemical studies, support a cooperation with transcriptional networks involved in urothelial differentiation. In collaboration with A. Losada ( CNIO ), we have developed a conditional Stag2 knockout strain and are analysing the role of cohesin in urothelial cell transformation. RBM10 codes for a splicing factor and it is mutated in several other epithelial tumours. Inactivation in UC is not associated with stage or grade, but it occurs mainly in tumours with Figure 1 Pancreatic KRasG 12V ;Gata4- null mice develop PanIN-less PDAC. Quantification of PanINs and PDAC in young (<40 weeks ) and old (>1 year ) mice ( A ). Acini from mice lacking Gata4 display reduced cyst formation upon culture in Matrigel. They also show a blunted ADM response to EGF but are able to respond to IL17 ( B ). SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 67 CANCER CELL BIOLOGY PROGRAMME | EPITHELIAL CARCINOGENESIS GROUP urothelial differentiation. In collaboration with J. Valcárcel ( CRG, Barcelona ), we have generated a conditional Rbm10 knockout strain and are analysing the molecular mechanisms through which this gene contributes to UC development using a combination of molecular and bioinformatics strategies. These studies will be complemented with the use of normal urothelial organoids, for which we have established robust culture methods and have shown their strict dependence on EGF and Wnt signalling. We have characterised an organoid cell-of-origin with stem cell properties in vitro and have identified conditions promoting urothelial differentiation ( FIGURE 2 ). In addition, we are expanding these studies to human bladder cancers. Within the context of a project funded by the Spanish Association Against Cancer ( AECC ), we are analysing the clinical usefulness of the new UC taxonomy. The main aim is to identify predictors of outcome and of response to cisplatin-based therapies in patients receiving perioperative chemotherapy. These studies are linked to the design of clinical trials that include molecular stratification criteria. This work is carried out in collaboration with N. Malats at the CNIO and the SOGUG cooperative group. s ∞ PUBLICATIONS ∞ Notta F et al. ( incl. Real FX ) ( 2016 ). A renewed model of pancreatic cancer evo- lution based on genomic rearrangement patterns. Nature 538, 378-382. ∞ Fernández LC, Torres M, Real FX ( 2016 ). Somatic mosaicism : on the road to cancer. Nat Rev Cancer 16, 43-55. ∞ Zhou W et al. ( incl. Real FX, Malats N ) ( 2016 ). Mosaic loss of chromosome Y is associated with common variation near TCL1A. Nat Genet 48, 563-568. ∞ Hedegaard J et al. ( incl. Real FX, Malats N ) ( 2016 ). Comprehensive Transcriptional Analysis of Early-Stage Urothelial Carci- noma. Cancer Cell 30, 27-42. ∞ Gubern A, Joaquin M, Marquès M, Maseres P, Garcia-Garcia J, Amat R, González- Nuñez D, Oliva B, Real FX, de Nadal E, Posas F ( 2016 ). The N-Terminal Phos- phorylation of RB by p38 Bypasses Its Inactivation by CDKs and Prevents Prolif- eration in Cancer Cells. Mol Cell 64, 25-36. ∞ Machiela MJ et al. ( incl. Real FX, Malats N ) ( 2016 ). Female chromosome x mo- saicism is age-related and preferentially affects the inactivated x chromosome. Nat Commun 7, 11843. ∞ Richart L, Carrillo-de Santa Pau E, Rio- Machin A, Pérez de Andrés M, Cigudosa JC, Sanchez-Arevalo Lobo VJ, Real FX ( 2016 ). BPTF is required for c-MYC tran- scriptional activity and in vivo tumorigen- esis. Nat Commun 7, 10153. ∞ Ferrer J, Real FX ( 2016 ). The cis-regu- latory switchboard of pancreatic ductal cancer. EMBO J 35, 558-560. ∞ Toll A, Fernández LC, Pons T, Groesser L, Sagrera A, Carrillo-de Santa Pau E, Vi- cente A, Baselga E, Vázquez M, Beltrán S, Pisano DG, Rueda D, Gut M, Pujol RM, Hafner C, Gut I, Valencia A, Real FX ( 2016 ). Somatic embryonic FGFR2 mutations in keratinocytic epidermal nevi. J Invest Dermatol 136, 1718-1721. ∞ Millán-Uclés A, Zuluaga S, Marqués M, Vallejo-Díaz J, Sanz L, Cariaga-Martínez AE, Real FX, Carrera AC ( 2016 ). E-cadherin downregulation sensitizes PTEN mutant cells. Oncotarget 7, 84054-84071. ∞ Mazarico JM, Sánchez-Arévalo Lobo VJ, Favicchio R, Greenhalf W, Costello E, Carrillo-de Santa Pau E, Marqués M, Lacal JC, Aboagye E, Real FX ( 2016 ). Choline Kinase Alpha ( CHKa ) as a Therapeutic Target in Pancreatic Ductal Adenocarci- noma : Expression, Predictive Value, and Sensitivity to Inhibitors. Mol Cancer Ther 15, 323-333. ∞ Masson-Lecomte A, López de Matura- na E, Goddard ME, Picornell A, Rava M, González-Neira A, Márquez M, Carrato A, Tardon A, Lloreta J, Garcia-Closas M, Silverman D, Rothman N, Kogevinas M, Allory Y, Chanock SJ, Real FX, Malats N ; SBC/EPICURO Study Investigators ( 2016 ). Inflammatory-Related Genetic Variants in Non-Muscle-Invasive Bladder Cancer Prognosis : A Multimarker Bayesian As- sessment. Cancer Epidemiol Biomarkers Prev 25, 1144-1150. ∞ López de Maturana E, Picornell A, Mas- son-Lecomte A, Kogevinas M, Márquez M, Carrato A, Tardón A, Lloreta J, García-Clo- sas M, Silverman D, Rothman N, Chanock S, Real FX, Goddard ME, Malats N ; SBC/ EPICURO Study Investigators ( 2016 ). Pre- diction of non-muscle invasive bladder cancer outcomes assessed by innova- tive multimarker prognostic models. BMC Cancer 16, 351. ∞ Lerner SP, McConkey DJ, Hoadley KA, Chan KS, Kim WY, Radvanyi F, Höglund M, Real FX ( 2016 ). Bladder Cancer Molecular Taxonomy : Summary from a Consensus Meeting. Bladder Cancer 2, 37-47. ∞ Richart L, Real FX, Sanchez-Arevalo Lobo VJ ( 2016 ). c-MYC partners with BPTF in human cancer. Mol Cell Oncol 3, e1152346. ∞ AWARDS AND RECOGNITION ∞ Scientific Advisory Board, Centre de Recherche des Cordeliers, Paris, France. Figure 2 Normal mouse urothelial organoids ( NMU-o ) express E-cadherin ( A ). Organoids have been passaged for >1 year. Quantification of organoid growth in a ‘ Leave-one-out growth factor ’ experiment ( B ). Expression of urothelial differentiation markers ( C ). Cd49f labels a population with stem cell properties ( D ). ANNUAL REPORT 2016 68 VICE-DIRECTION OF BASIC RESEARCH EPITHELIAL CELL BIOLOGY JUNIOR GROUP OVERVIEW Tumour cells evolve into a progressively complex interplay between heterogeneous tumour cells and their tissue macroenviroment, which influences their proliferation and malignancy. Identifying the signalling mechanisms and cell types that sustain this complexity is one of the major goals in cancer biology. In adult skin, epithelial progenitor cells have been identified as the cell of origin of skin carcinomas. Several studies have been instrumental for defining regulatory pathways controlling their proliferation and/or differentiation. However, the identification of extrinsic factors modulating stem cell behaviour has not progressed very far to date. Using skin as a model system and employing mouse genetics and human samples, our research aims to understand how the interactions between epithelial progenitor cells, and also the interactions with their surrounding macroenvironment, sustain skin homeostasis, regeneration, and when perturbed lead to cancer. This information may provide insights for the future development of regenerative and anti-cancer therapies. Mirna Pérez-Moreno Junior Group Leader Post-Doctoral Fellow Silvia M. Janeiro “ During 2016, we continued our efforts to uncover novel events controlling the behaviour of skin stem cells in order to open up new insights into the mechanisms that control their regenerative characteristics, and how when disrupted they can lead to cancer.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 69 CANCER CELL BIOLOGY PROGRAMME | EPITHELIAL CELL BIOLOGY JUNIOR GROUP RESEARCH HIGHLIGHTS Regulation of epidermal progenitor cells self-renewal and differentiation During 2016, we continued exploring how tissues acquire an adequate control of cell division and differentiation. In particular, using mouse epidermal development as a model system, we investigated the contributions of mitotic and cytoskeletal proteins in the regulation of skin progenitors ’ self-renewal through oriented cell divisions. Contributions of stromal cells to the skin stem cell niche in homeostasis We have recently identified a novel connection between macrophages and skin progenitor cells, which modulates their stem cell properties and regenerative potential. We are expanding these results to decipher how other signals, and cells from the stroma, are connected with the skin stem cell niche and regulate skin regeneration. Contributions of stromal cues in cancer stem cell maintenance and tumour progression The formation of tumours and their progression to malignancy undoubtedly involves the contributions of the tumour macroenvironment. Identifying the signalling mechanisms and cell types that contribute to tumour initiation and progression to malignancy is instrumental for detecting potential targets for clinical applications aimed at eradicating tumours. The macroenvironment of many tumours is rich in cytokines, chemokines, and inflammatory enzymes. During 2016, we continued exploring the role of diverse cell-derived soluble mediators in modulating proliferation, migration and survival of skin cancer stem cells. In addition, we focused our efforts on dissecting the contributions of immune cells to the cancer stem cell niche in tumour initiation and development. We are employing conditional loss- and gain- of-function studies in genetically modified mice in order to demonstrate the role of specific cell types and their derived soluble mediators in tumourigenesis ; this may provide further insights for the potential development of immunotherapeutic approaches. s. Graduate Student Daniel Peña Technician Francesca Antonucci ( TS )* ( until March ) *Titulado Superior ( Advanced Degree ) Figure Skin carcinoma showing the presence of a high density of inflammatory cells within the tumour. Inset shows a magnification of immune infiltrates within the tumour. *Arrows point to some immune cells ANNUAL REPORT 2016 70 VICE-DIRECTION OF BASIC RESEARCH GROWTH FACTORS, NUTRIENTS AND CANCER JUNIOR GROUP OVERVIEW The incidence of metabolic disease and cancer has increased to epidemic proportions, possibly due to hypernutrition and a more sedentary life style with less energy expenditure. Our laboratory studies the molecular mechanisms of disease associated with the deregulation of growth factor and nutrient signalling pathways. Identifying new components of the growth factor and nutrient cascades, as well as elucidating their role and functions in vivo by generating new genetically engineered mouse models ( GEMMs ), will help us to better understand how nutrient overload can induce metabolic disorders and cancer. Thus, using cell biological and biochemical techniques, combined with in vivo mouse models and human data, our lab devotes efforts to the development of innovative mechanism-based therapeutics to potentially treat metabolic dysfunctions and cancer. “ Our research focus is to generate mouse models recapitulating human disease associated to nutrient overload in order to guide research perspectives and applications.” Research concepts from our laboratory ɗ Metabolic alterations initiate tumorigenesis prior to genomic instability. ɗ Inhibition of de novo NAD+ synthesis functions a non- oncogene addiction pathway in liver and pancreas cancer. ɗ Oncogene-induced NAD+ depletion in DNA damage. Junior Group Leader Nabil Djouder Post-Doctoral Fellow Hugo Bernard SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 71 CANCER CELL BIOLOGY PROGRAMME | GROwTH FACTORS, NUTRIENTS AND CANCER JUNIOR GROUP RESEARCH HIGHLIGHTS We have a particular interest in studying gastrointestinal track disorders. Our work in this area focuses on metabolic organs such as the liver, intestine and pancreas, as these 3 organs are physiologically interconnected and influenced through their exocrine and/or endocrine functions and microbiota ( FIGURE ). Our task is thus to generate new mouse models mimicking human disease and to study mechanisms and events iniating disease development. We also use patient-derived xenograft models and organoids to translate our findings into clinical perspectives. Guided by experimental mouse models combined with the use of human data, we aim to provide a comprehensive study for a rational approach towards the development of novel mechanism-based therapeutics to prevent, ameliorate and treat diseases. Identifying new components of growth factor and nutrient signalling cascades We identified 2 components of the growth factor and nutrient signalling cascades regulating the mTORC1 pathway : Uncoventional prefoldin RPB5 interactor ( URI ) ( Djouder et al., 2007 ) and Microspherule protein 1 ( MCRS1 ) ( Fawal et al., 2015 ). Uncoventional prefoldin RPB5 interactor ( URI ): URI is member of the R2TP/URI-prefoldin like complex, which contains not only prefoldin subunits but also RNA polymerase binding subunit ( RPB5 ), ATPases/helicases RuvB-like protein 2 ( RUVBL2, also known as 48- kDa TATA box-binding protein- interacting [ TIP48 ] or reptin ) and RuvB-like protein 1 ( RUVBL1, also known as 49-kDa TATA box-binding protein- interacting [ TIP49 ] or pontin ) and co-chaperones such as heat shock protein 90 ( HSP90 ). URI is a downstream component of the growth factor and nutrient signalling pathways. It is phosphorylated by S6K1 and has an oncogenic role in ovarian cancer and HCC development. Microspherule protein 1 ( MCRS1 ): MCRS1, in an amino acid- dependent manner, maintains Rheb at lysosome surfaces, connecting Rheb to mTORC1. MCRS1 depletion promotes Rheb/ TSC2 interaction, rendering Rheb inactive and delocalising it from lysosomes to recycling endocytic vesicles, leading to mTORC1 inactivation. Generation of genetically engineered mouse models ɗ 2 conditional knock-outs ( URI and MCRS1 loss-of-function ). ɗ 3 knock-ins ( over-expression of URI ( wt ), URI ( S371A ) and MCRS1 ). Research achievements ɗ Inflammatory cues and nutrient overloads up-regulate hepatic URI. ɗ URI is an oncogene initiating NASH and HCC. ɗ Nicotinamide riboside to prevent liver and pancreas cancers. ɗ MCRS1 activates mTORC1 in response to amino acids. ɗ URI is the first identified OGT regulator in response to glucose fluctuations. ɗ Glucose depletion can induce oncogenic signals through OGT/c-MYC regulation. ɗ c-MYC is oncogenic and tumour suppressive depending on nutrient availability. s Graduate Students Marta Brandt, Almudena Chaves, Amanda Garrido, Ana Teijeiro Technician Eva Martínez ( since March ) ( PEJ )* Student In Practice Tatiana Grazioso ( Universidad Complutense de Madrid ) *Plan de Empleo Joven ( Youth Employment Plan ) ∞ PUBLICATIONS ∞ Burén S, Gomes AL, Fawal MA, Teijeiro A, Yilmaz M, Tummala KS, Perez M, Rodri- guez-Justo M, Campos-Olivas R, Megias D, Djouder N ( 2016 ). Regulation of OGT by URI in Response to Glucose Confers c-MYC-Dependent Survival Mechanisms. Cancer Cell 30, 290-307. ∞ Gomes AL, Teijeiro A, Burén S, Tummala KS, Yilmaz M, Waisman A, Theurillat JP, Perna C, Djouder N ( 2016 ). Metabolic Inflammation-Associated IL17A Causes Non Alcoholic Steatohepatitis and Hepa- tocellular Carcinoma Development. Cancer Cell 30, 161-175. ∞ Djouder N ( 2016 ). Adaptive survival mech- anism to glucose restrictions. Oncoscience 3, 302-303. ∞ Garrido A, Brandt M, Djouder N ( 2016 ). Transport to Rhebpress Activity. Small GTPases 7, 12-15. Figure Milestones in growth factors and nutrients research in my laboratory. The scheme illustrates our present and future research. Time and effort are dedicated to better understand how misregulation in growth factor and nutrient signalling pathways can lead to gastrointestinal track disease development. ANNUAL REPORT 2016 72 VICE-DIRECTION OF BASIC RESEARCH SEVE BALLESTEROS FOUNDATION-CNIO BRAIN TUMOUR JUNIOR GROUP OVERVIEW Malignant gliomas ( astrocytomas, oligodendrogliomas and oligoastrocytomas ) are the most frequent form of brain tumours and Glioblastoma Multiforme ( GBM ), a grade IV astrocytoma, is the most lethal tumour of the central nervous system in the adult. Standard GBM therapy consists of tumour resection and postsurgical treatment with chemotherapy and ionising radiation ( IR ). Although there have been improvements in surgical and imaging techniques, available treatments for GBMs are still inefficient, most likely due to intrinsic resistance to the current therapeutic modalities and high cellular heterogeneity. In our laboratory, we use a combination of genomic analysis, mouse models and primary tumour cell cultures, with the main goal of identifying the molecular mechanisms that could provide the basis for novel treatments for GBM patients. “ The main focus of our Group is to uncover the genetic alterations present in GBM patients that are responsible for the aggressiveness and the poor treatment response of this tumour type.” Massimo Squatrito Junior Group Leader Staff Scientists Bárbara Oldrini, Alberto J. Schuhmacher SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 73 CANCER CELL BIOLOGY PROGRAMME | SEVE BALLESTEROS FOUNDATION-CNIO BRAIN TUMOUR JUNIOR GROUP RESEARCH HIGHLIGHTS Next generation glioma mouse models A decade of studies has underlined the complexity of the glioma genome, however, the functional significance of the vast majority of the genetic alterations remains elusive. Understanding the genetic events that lead to glioma formation and the mechanisms of resistance to therapy will be instrumental for the development of new treatment modalities for gliomas. To accurately reproduce the high genetic heterogeneity observed in glioma patients, we would have to recreate not just a handful of genetic alterations, but possibly dozens. The advent of the CRISPR/Cas genome editing technology has now made it possible to target almost any candidate cancer gene in the in vivo setting. We are actively working to develop the ‘ next-generation ’ glioma mouse models that more faithfully recapitulate in vivo the complexity of the GBM genome, with a particular interest in tumour suppressor genes and complex gene rearrangements. Overcoming therapy resistance in GBM The standard therapies for GBM patients, IR and temozolomide ( TMZ ), generate double-stranded DNA breaks ( DSDBs ), the most deleterious form of DNA damage. The DSDBs are then responsible for the initiation of the DNA Damage Response ( DDR ) and, consequently, the activation of DNA repair pathways and cell-cycle checkpoints. DDR signalling is a very intricate pathway and many of its elements can be altered in a given tumour patient, offering both challenges and opportunities from a treatment perspective. The most frequent resistance mechanism to TMZ treatment is the expression of the DNA- repair gene O 6-methylguanine DNA methyltransferase ( MGMT ), however, other resistance mechanisms have still to be identified. Through a variety of genetic approaches ( Haploid cells transposone mutagenesis, gRNA and shRNA screenings ) we have identified the main signalling pathways that mediate resistance to TMZ. We are currently performing a series of synthetic lethality screenings in order to bypass these mechanisms of resistance. s Graduate Students Carolina Almeida, Alvaro Curiel, Veronica Matía ( since October ) Technician Claudia S. Troncone Students in Practice Paula Nogales ( until July ), Anna Salamero ( until July ) ∞ PUBLICATIONS ∞ Bowman R, Wang Q, Carro A, Verhaak RGW, Squatrito M ( 2016 ). GlioVis data portal for visualization and analysis of brain tumor expression datasets. Neu- ro-Oncology. PMID : 28031383. ∞ de Lucas AG *, Schuhmacher AJ *, Oteo M, Romero E, Cámara JA, de Martino A, Arroyo AG, Morcillo MÁ, Squatrito M#, Martinez-Tor- recuadrada J#, Mulero F# ( 2016 ). Targeting MT1-MMP as an immunoPET-based strategy for imaging gliomas. PloS One 11, e0158634. *Co-first author, #corresponding author ∞ AWARDS AND RECOGNITION ∞ Alberto J. Schuhmacher has been awarded a Research Contract from the Ramon y Ca- jal Programme ( funded by the Ministerio de Economía, Industria y Competitividad, MEIC ), and the “ Eduardo Gallego ” Grant of the Francisco Cobos Foundation, Spain. ∞ Álvaro Curiel has received the Beca de Honor - Colegio Mayor Larraona given by the University of Navarra, Spain. Figure Schematic representation of the RCAS-CRISPR-Cas9 system to generate gliomas with tailored genetic alterations. ANNUAL REPORT 2016 74 VICE-DIRECTION OF BASIC RESEARCH STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME ALFONSO VALENCIA Programme Director SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 75 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME The objective of the Structural Biology and Biocomputing Programme is the mechanistic understanding of key cancer related molecular systems. The Programme was designed to combine computational and structural approaches, and to collaborate with the CNIO Basic and Translational Research activities. Our 3 main research goals are to : ɗ Reconstruct the structural details of protein complexes that are active in cancer and related processes. ɗ Predict the consequences of cancer related alterations ; we are focusing on alterations of compensatory nature ( co-evolutionary related mutations ) as well as those affecting alterative splicing patterns. ɗ Contribute to the analysis of cancer epigenomic and genomic information as part of international genome projects. Currently the Programme includes 3 Research Groups and 6 Core Units that provide support to the CNIO’s research activities. Following the recommendations of the CNIO’s External Scientific Advisory Board ( SAB ), resulting from the 2015 review of the Structural Biology and Biocomputing Programme, we started the process of recruiting additional Groups for the Programme. The selection of candidates for two Junior Group Leader positions was carried out with the help of an ad-hoc external and an internal selection committee. Six outstanding candidates were invited to visit the CNIO, defend their work and have a discussion with the corresponding committees. Finally, two candidates were selected covering the areas of biochemistry-structural biology and electron microscopy - protein complexes. These two new Groups will start their work at the CNIO in the first part of 2017. Once these Groups are consolidated, the plan is to re-evaluate the possibility of hiring a senior crystallographer as recommended by the SAB. On the computational side, the Programme has seen the departure of the Heads of the Bioinformatics and the National Bioinformatics Institute ( INB-ISCIII ) Units ; they have since been replaced by Salvador Capella, as Head of the INB-ISCIII/ ELIXIR Unit, and by Fátima Al-shahrour, who will coordinate the Bioinformatics Unit and absorb the activities of the previous Translational Bioinformatics Unit. Of particular relevance for the CNIO’s activities in Computational Biology, was the finalisation of the negotiation with the Ministerio de Energía, Turismo y Agenda Digital for the implementation of a biological text mining platform in the framework of the ‘ Plan de Impulso de las Tecnologías del Lenguaje ’. Within the CNIO structure, this activity will fall under and be developed by a new ‘ Text Mining ’ Unit headed by Martin Krallinger. This Unit will start operations at the beginning of 2017. Despite these positive developments, the CNIO still needs to reinforce the computational side of the Programme ; particularly, research related to the Experimental Therapeutics Programme as well as Computational Cancer Genomics needs to be further strengthened. “ The Programme is about to undergo an important transition with the addition of two new Groups. Several very positive new activities in the area of data and text mining have also come about. The establishment of the new Groups and the reinforcement of the structural and computational activities at the CNIO remain challenges for the coming year.” ANNUAL REPORT 2016 76 VICE-DIRECTION OF BASIC RESEARCH STRUCTURAL COMPUTATIONAL BIOLOGY GROUP Alfonso Valencia Group Leader Staff Scientists Andrea Nicole Dölker, Vera Pancaldi, Tirso Pons, Daniel Rico ( until July ), Michael Tress SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 77 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME | STRUCTURAL COMPUTATIONAL BIOLOGY GROUP OVERVIEW The main interest of our Group is the study of the molecular bases of cancer by bringing an evolutionary perspective to the study of the interplay between genomics and epigenomics in tumour progression. Our research is largely carried out in the context of large-scale genome projects, in which we develop new computational methods for the study of genome-cancer relationships. In this general scenario, the strategic goals of the Structural Computational Biology Group are to : ɗ Develop new ideas, methods and software platforms for the extraction, integration and representation of cancer data, including the analysis of molecular, genomic, epigenomic and phenotypic information in collaboration with large-scale genome projects. ɗ Include new technologies for data and text mining, together with Machine Learning methods, in our cancer genome analysis framework. ɗ Analyse the function, structure and specific interactions of cancer- related proteins. “ This year the initial phase of two large scale projects was completed ; i.e. the International Human Epigenome Consoritum ( iHEC ) and the Pancancer Analysis of Whole Genomes ( PCAWG ). In both cases, we have contributed to the computational analysis, including the implementation of the data analysis infrastructures and the development of new analysis methods, as well as collaborating towards the interpretation of the biological results.” RESEARCH HIGHLIGHTS The Group has contributed to several community efforts in different areas : ɗ Epigenomics with the BLUEPRINT EU flagship project, which is part of the iHEC consortium ; the results from this work were published at the end of 2016. ɗ Pancancer Analysis of Whole Genomes ( PCAWG ), global analysis of 2500 complete cancer genomes ; these results will be published in 2017. ɗ The BioCreative text mining challenge in chemical compounds resulted in a number of resources and publications that appeared throughout 2016. We have introduced a new computational method for the prediction of pairs of residues in protein interfaces. This method can help in the analysis of cancer related mutations. We have also introduced new methods for the analysis of epigenomes at the linear two dimensional level ( chromatin states ) and three dimensional level ( chromatin structure in the nucleus ). The cancer genome analysis system Our Group is deeply involved in the development of a computational framework for the analysis of human genomes with specific application to the analysis of cancer genomes. Over the years, this framework has been applied to a number of collaborative cancer projects, and it has been particularly instrumental in the CLL-ICGC project. We have now moved on to a new phase in which the framework is used for the analysis of the large set of full cancer genomes Post-Doctoral Fellows Simone Marsili ( until October ), Miguel Vázquez ( until September ) Graduate Students Maria Rigau, Juan Rodríguez, Jon Sánchez Technicians David A. Juan ( TS )*, Martin Krallinger ( TS )*, Miguel Madrid ( since November ) ( TS )*, Filipe N. Were ( TS )* *Titulado Superior ( Advanced Degree ) Visiting Scientists Dimitrios Morikis ( University of California, Riverside, USA ), Miguel Vazquez ( Norwegian University of Science and Technology, Trondheim, Norway ) ANNUAL REPORT 2016 78 VICE-DIRECTION OF BASIC RESEARCH of the Pancancer Analysis of Whole Genomes ( PCAWG ); it is one of the four frameworks for data organisation, analysis and exploration used by the consortium. With regards to the future, given the characteristics of the framework in terms of its modular structure, capacity of integration of new methods in working pipelines, and ease of installation ( e.g. adoption of docker and cloud technologies ), we consider that it can be the seed of new developments in the overarching analysis of human disease genomes. Protein structure prediction and cancer genomes In the context of cancer genome analysis, and as part of the Pan Cancer global effort, we have developed a set of methods to analyse the consequences of mutations in the interface of proteins. The underlying logic is that cellular functions are governed by signals transmitted via protein interactions and protein complexes. In these interactions, the amino acids located in interacting surfaces determine the intensity of the interactions and, very importantly, the specificity of the interactions. The exquisite functioning of cellular systems between proteins depends critically on the pairing of the proteins with their correct partners, and the accuracy of the interactions depends on the correct formation of pairs of residues of the 2 proteins in the interface. We have shown that cancer associated mutations tend to accumulate in the protein interfaces to the point that, with the information available, it is possible to say that cancer related  mutations specifically target protein interfaces. Therefore,  understanding the nature of protein-protein interactions is important for understanding the impact of cancer mutations. We have developed a new methodology able to predict, with high accuracy, a small set of pairs of residues located in the interface of interacting human proteins. The new methodology, based on the study of the co-evolution of the corresponding protein families, does not require any information about the corresponding structures and it is applicable to many human protein complexes for which no other information is available. Furthermore, we have shown that the pairs of residues predicted to interact are very conserved in structural terms ( they occupy the same position in space over the lengthy evolutionary time ), which is indicative of their importance in the organisation of the corresponding interfaces. Based on these results, we are now exploring the use of the newly developed computational methods as an alternative approach for the interpretation of the consequences of cancer related mutations. EPIGENOME analysis infrastructure and portal In the context of the BLUEPRINT iHEC project we have designed a system for the comparative analysis of epigenetics data ( the BluePrint analysis portal http ://blueprint-data.bsc.es/ release_2016-08/, developed in collaboration with the BSC-CNS and EBI-EMBL ). This portal is now the main point of access for the project’s results ( e.g. chromatin states, ChIP-Seq positions of histone modifications ), enabling the direct comparison of the epigenetic structure of different cell types. Based on the information provided by the Blueprint Analysis Portal, we have developed the methodology to compare epigenomes at the level of their organisation in functional segments ( chromatin states ). The initial results show that the system is not only able to reproduce the structure of the lineage differentiation during haematopoiesis, but also to detect what the main potential epigenetic driving factors of the differentiation are. The method, initially developed for the Blueprint data sets, is now being extended to other data types provided by the iHEC consortium. Alternative splicing at the protein level In 2016, we continued our work on alternative splicing in the context of the NIH-funded GENCODE project. Our results, summarised in a review published in TIBS ( Tress et al., 2016 ), show that in light of combined approaches, including protein modelling, proteomics and evolutionary analysis, there is little evidence to demonstrate that alternative isoforms are expressed at the protein level in detectable quantities. In other words, the only available evidence is that normal proteins are coded by the principal isoform of each gene and not by any of the potential alternative forms that are undoubtedly produced at the mRNA level. Even if this observation is in line with recent results of the large scale analysis of gene expression in human tissues ( publications of the ENCODE/GTEx -www.gtexportal.org ), it is still somewhat controversial since it indicates a big unexplained discrepancy Figure 1 Co-evolution based correct prediction of pairs of residues in the interface of 2 domains of the human cytosolic phenylalanine tRNA synthetase (a subunit in dark red, B5 and B3/4 domains in β subunit in purple and dark blue, respectively); taken from Rodriguez-Rivas et al., 2016 . SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 79 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME | STRUCTURAL COMPUTATIONAL BIOLOGY GROUP between the results obtained at the level of gene and protein expression ; a discrepancy that might have profound implications for our understanding of the role of mRNA in cells and the overall understanding of the biological function of processed RNAs. Biological Text Mining Text mining, an important part of the Group’s activity, has broad implications in Biomedicine. In 2016, we completed this year an exhaustive review of the application of text mining to the area of chemistry ( Krallinger et al., this work has been submitted to Chem Rev ); this review was based on our experience in the analysis of text mining systems and the results in the context of the 2015 BioCreative Chemdner challenge ( http ://www. biocreative.org/tasks/biocreative-iv/chemdner/). During 2016, we reached an agreement with the Ministerio de Energía, Turismo y Agenda Digital for the implementation of a biological text mining platform in the framework of the ‘ Plan de Impulso de las Tecnologías del Lenguaje ’; this project is to develop tools and procedures in line with the recommendations of the European e-Infrastructure in text mining OpenMinted, in which we also participate. s ∞ PUBLICATIONS ∞ Astle WJ et al. ( incl. Valencia A ) ( 2016 ). The Allelic Landscape of Human Blood Cell Trait Variation and Links to Common Complex Disease. Cell 167, 1415-1429. ∞ Chen L et al. ( incl. Valencia A ) ( 2016 ). Drivers of Epigenetic and Transcriptional Variation in Human Immune Cells. Cell 167, 1398-1414. ∞ Stunnenberg HG ; International Human Epigenome Consortium., Hirst M ( 2016 ). The International Human Epigenome Con- sortium : A Blueprint for Scientific Collab- oration and Discovery. Cell 167, 1145-1149. ∞ Tress ML, Abascal F, Valencia A ( 2016 ). Alternative Splicing May Not Be the Key to Proteome Complexity. Trends Biochem Sci. PMID : 27712956. ∞ Rodriguez-Rivas J, Marsili S, Juan D, Valencia A ( 2016 ). Conservation of coevolving protein interfaces bridges prokaryote-eukaryote homologies in the twilight zone. Proc Natl Acad Sci USA 113, 15018-15023. ∞ Galindo-Albarrán AO et al. ( incl. Va- lencia A ) ( 2016 ). CD8+ T Cells from Human Neonates Are Biased toward an Innate Immune Response. Cell Rep 17, 2151-2160. ∞ Juan D, Perner J, Carrillo de Santa Pau E, Marsili S, Ochoa D, Chung HR, Vingron M, Rico D, Valencia A ( 2016 ). Epigenomic Co-localization and Co-evolution Reveal a Key Role for 5hmC as a Communication Hub in the Chromatin Network of ESCs. Cell Rep 14, 1246-1257. ∞ Toll A, Fernández LC, Pons T, Groesser L, Sagrera A, Carrillo-de Santa Pau E, Vi- cente A, Baselga E, Vázquez M, Beltrán S, Pisano DG, Rueda D, Gut M, Pujol RM, Hafner C, Gut I, Valencia A, Real FX ( 2016 ). Somatic embryonic FGFR2 mutations in keratinocytic epidermal nevi. J Invest Dermatol 136, 1718-1721. ∞ Abascal F et al. ( incl. Valencia A ) ( 2016 ). Extreme genomic erosion after recurrent demographic bottlenecks in the highly endangered Iberian lynx. Genome Biol 17, 251. ∞ Jiang Y et al. ( incl. Valencia A ) ( 2016 ). An expanded evaluation of protein function prediction methods shows an improve- ment in accuracy. Genome Biol 17, 184. ∞ Pancaldi V, Carrillo-de-Santa-Pau E, Javierre BM, Juan D, Fraser P, Spivakov M, Valencia A, Rico D ( 2016 ). Integrating epigenomic data and 3D genomic struc- ture with a new measure of chromatin assortativity. Genome Biol 17, 152. ∞ Gurard-Levin ZA, Wilson LO, Pancaldi V, Postel-Vinay S, Sousa FG, Reyes C, Ma- rangoni E, Gentien D, Valencia A, Pommier Y, Cottu P, Almouzni G ( 2016 ). Chromatin regulators as a guide for cancer treatment choice. Mol Cancer Ther 15, 1768-1777. ∞ Berger B, Gaasterland T, Lengauer T, Orengo C, Gaeta B, Markel S, Valencia A ( 2016 ). ISCB’s Initial Reaction to The New England Journal of Medicine Editorial on Data Sharing. PLoS Comput Biol 12, e1004816. Bioinformatics. PMID:27153698. & F1000Res 5, pii: ISCB Comm J-157. ∞ Pons T, Vazquez M, Matey-Hernandez ML, Brunak S, Valencia A, Izarzugaza JM. ( 2016 ). KinMutRF : a random forest classifier of sequence variants in the human protein kinase superfamily. Bmc Genomics 17, 396. ∞ Pérez-Pérez M, Pérez-Rodríguez G, Rabal O, Vazquez M, Oyarzabal J, Fdez-Riverola F, Valencia A, Krallinger M, Lourenço A ( 2016 ). The Markyt visualisation, predic- tion and benchmark platform for chemical and gene entity recognition at BioCre- ative/CHEMDNER challenge. Database ( Oxford ), pii : baw120. ∞ Durinx C, McEntyre J, Appel R, Apweiler R, Barlow M, Blomberg N, Cook C, Gasteiger E, Kim JH, Lopez R, Redaschi N, Stock- inger H, Teixeira D, Valencia A ( 2016 ). Identifying ELIXIR Core Data Resources. F1000Res 5, pii : ELIXIR-2422. ∞ Fernández JM, de la Torre V, Richard- son D, Royo R, Puiggròs M, Moncunill V, Fragkogianni S, Clarke L ; BLUEPRINT Consortium., Flicek P, Rico D, Torrents D, Carrillo de Santa Pau E, Valencia A ( 2016 ). The BLUEPRINT Data Analysis Portal. Cell Syst 3, 491-495. Figure 2 Representation of chromatin state transitions during haematopoietic differentiation. Boxes represent chromatin states and lines represent the observed transitions between cell types. ANNUAL REPORT 2016 80 VICE-DIRECTION OF BASIC RESEARCH OVERVIEW Our Group studies regulatory mechanisms of key signalling switches controlling growth and adhesion signals. Such signals regulate important cellular processes such as proliferation, adhesion and survival. We use structural techniques, such as X-ray crystallography and electron microscopy, in combination with biochemical and functional studies to understand these mechanisms at atomic detail and to rationalise how oncogenic events result in their deregulation. The structural understanding allows us to design potential anti-cancer therapeutics that interfere with oncogenic deregulation. We focus on mechanisms of growth and adhesion signalling that occur at the plasma membrane and involve specific phosphoinositides. In particular, we aim to answer two main questions : ( i ) what are the events occurring at the cell membrane at integrin adhesion sites that trigger Focal Adhesion Kinase signalling ; and ( ii ) how are phosphatidylinositol ( 3,4,5 )-trisphosphate ( PIP3 ) levels regulated to affect signalling of the Akt pathway. CELL SIGNALLING AND ADHESION JUNIOR GROUP Daniel Lietha Junior Group Leader Post-Doctoral Fellows Iván Acebrón, Johanne Le Coq “ We obtain detailed structural and mechanistic insights in order to understand how growth and adhesion signals are triggered to cause tumour invasion, and we use this information for allosteric targeting.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 81 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME | CELL SIGNALLING AND ADHESION JUNIOR GROUP RESEARCH HIGHLIGHTS We showed that Focal adhesion Kinase ( FAK ) interacts with PIP2 lipids at cell adhesion sites and that this interaction induces FAK clustering and conformational changes, which trigger FAK autophosphorylation. Following this, Src is recruited to FAK and, in turn, phosphorylates the FAK kinase to induce full FAK activation. Currently, we are studying the architecture of FAK clusters bound to lipid membranes by electron microscopy, and we are investigating how force, induced at adhesion sites by actomyosin contraction, induces changes to these structures in order to activate focal adhesion signalling. We utilise these mechanistic insights to discover highly specific allosteric FAK inhibitors. We employ experimental and virtual screening, using fragment based approaches, to identify allosteric ligands, and then use a structure based approach to develop these fragments into inhibitory lead compounds. SH2-domain-containing inositol 5-phosphatases ( SHIP ) remove the 5-phosphate from PIP3 and thereby, like PTEN, negatively regulate PIP3 levels. Despite their importance, little is known about the mechanisms of SHIP regulation. We solved a crystal structure containing the catalytic and C2 domains of SHIP2, showing an extensive interface between the two domains. We have shown that the C2 domain of SHIP2 binds phosphatidylserine, and hence the rigid C2 interaction efficiently positions the active site towards its substrate ( FIGURE ). Although the C2 domain interacts with the phosphatase domain far from the active site, we show that the C2 interaction greatly enhances the catalytic activity of SHIP2. We employed molecular dynamics simulations to guide a mutagenesis study that has identified distinct allosteric signalling pathways emanating from hydrophobic or polar interdomain interactions, which differentially affect lipid chain or head group regions of the substrate. Furthermore, we confirmed via cell biology experiments that mutations at the domain interface affect downstream signalling to Akt. s Graduate Students Marta Acebrón, Marta Camacho ( until May ), José Vicente Velázquez Technician Pilar Redondo ∞ PUBLICATIONS ∞ Lietha D ( 2016 ). Forcing FAK into Tran- scriptional Activity. Structure 24, 1223-1225. ∞ Le Coq J, Heredia Gallego L, Lietha D ( 2016 ). Expression, Purification, Crystalli- sation and X-ray Crystallographic Analysis of a Truncated Form of Human Src Homol- ogy 2 Containing Inositol 5-Phosphatase 2. Protein J 35, 225-230. Figure Using a multidisciplinary approach we defined two allosteric paths emanating from the interface between the phosphatase ( 5-Ptase ) and C2 domains of SHIP2, which via hydrophobic regions (‘ H-path ’) or a connection of polar residues (‘ P-path ’) affect different substrate binding regions in the active site. ANNUAL REPORT 2016 82 VICE-DIRECTION OF BASIC RESEARCH STRUCTURAL BASES OF GENOME INTEGRITY JUNIOR GROUP OVERVIEW Safeguarding genome integrity is essential for correct cell functioning and to prevent cancer. Our Group is interested in understanding central cellular processes that affect the integrity of the genome, such as the metabolism of nucleotides, DNA recombination or the maintenance and recognition of chromatin architecture. These processes depend on the assembly of large and dynamic macromolecular complexes. We combine protein engineering, X-ray crystallography, nuclear magnetic resonance ( NMR ) and single-particle electron microscopy ( EM ), together with biochemical and functional studies, in order to decipher the structure of these protein-protein and protein-DNA complexes, as well as to understand their catalysis and regulatory mechanisms at the atomic level. This knowledge should guide the design of compounds to modulate protein activity and provide novel opportunities for fighting tumours. “ We obtained an atomic view of the ATC domain of human CAD – a metabolic gatekeeper controlling cell proliferation – bound to the anti-tumour drug PALA, and localised CAD within the cell.” Santiago Ramón-Maiques Junior Group Leader Post-Doctoral Fellow Maria Dolores Moreno SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 83 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME | STRUCTURAL BASES OF GENOME INTEGRITY JUNIOR GROUP RESEARCH HIGHLIGHTS Unmasking CAD, a metabolic gatekeeper of cell proliferation CAD is a 1.5 MDa multi-enzymatic complex formed by hexameric association of a ~240 kDa polypeptide with four functional domains : glutaminase ( GLNase ), carbamoyl phosphate synthetase ( CPSase ), aspartate transcarbamoylase ( ATCase ) and dihydroorotase ( DHOase ). Each domain catalyses one of the initiating steps in the de novo biosynthesis of pyrimidine nucleotides. CAD is tightly regulated by allosteric effectors and by phosphorylation through different signalling cascades, and its activity is key to fuel the high demand of pyrimidines during cell growth and proliferation. Despite its central role in metabolism and its potential as an anti-tumour target, there is no detailed information about the architecture of CAD or about the structure of any of its functional domains. We aim to decipher the structure of the complex and to understand its catalytic and regulatory mechanisms at the atomic level. Structure and functioning of the ATC domain of human CAD We resolved the crystal structure of the ATCase domain of human CAD – free or bound to carbamoyl phosphate, or to the anti-tumour drug PALA – confirming its overall similarity with bacterial homologues ( Ruiz-Ramos et al., 2016 ). Unexpectedly, we found a decreasing affinity for PALA that could help to understand tumour resistance to this drug. Mutagenic and biochemical analysis linked the lowered PALA affinity to the communication of conformational changes between the ATCase subunits. The mutation of one key residue in this mechanism was recently found by others to cause the first CAD-related human disease ( Ng B.C. et al., Hum Mol Genet, 2015 ). Using CRISPR to understand the functioning of CAD in vivo We generated fluorescent recombinant chimeras and used CRISPR to introduce green fluorescent protein ( GFP ) into the endogenous CAD gene and to knockout CAD in human cell lines. These tools enable us to interrogate important aspects of CAD functioning in vivo. By tracking the subcellular localisation of CAD in mammalian cells we demonstrated that, contrary to previous reports, CAD is located exclusively at the cytosol and does not translocate into the nucleus during the cell cycle. These engineered proteins and gene edited cells are also proving to be instrumental for the identification of interacting protein partners and for the testing of the disease- causing potential of newly identified clinical mutations in CAD. s Graduate Students Francisco Del Caño, Alba Ruiz ( until February ) Technicians Araceli Grande ( TS )*, Igor Yefimenko ( TS )* *Titulado Superior ( Advanced Degree ) ∞ PUBLICATION ∞ Ruiz-Ramos A, Velázquez-Campoy A, Grande-García A, Moreno-Morcillo M, Ramón-Maiques S ( 2016 ). Structure and functional characterization of human as- partate transcarbamoylase, the target of the anti-tumoral drug PALA. Structure 24, 1081-1094. Figure ( A ) Cartoon representation of human ATCase trimer. ( B ) Crystal structure of human ATCase bound to the anti-tumour drug PALA. ( C, D ) Subcellular localisation of CAD using fluorescence microscopy in U2-OS wild- type cells ( C ) and in CRISPR-generated CAD knock out cells ( D ). C DBA ANNUAL REPORT 2016 84 VICE-DIRECTION OF BASIC RESEARCH SPECTROSCOPY AND NUCLEAR MAGNETIC RESONANCE UNIT OVERVIEW The Unit unifies the technical and scientific management of Nuclear Magnetic Resonance Spectroscopy ( NMR ) and of other biophysical instrumentation available through the Structural Biology and Biocomputing Programme. It provides CNIO researchers with instrumentation and technical support for a variety of spectroscopic and biophysical techniques. This includes the application of NMR to the in vitro characterisation of the structure and dynamics of biomolecules ( proteins in particular ) and their interactions with other biopolymers, as well as with small molecules that could represent initial hits in the drug discovery process or research compounds for biophysical and functional studies. Furthermore, we use NMR to characterise the metabolic profiles of biofluids, cell growth media and cell and tissue extracts from both animal models of cancer and human samples. “ In 2016, we quantified metabolites from cell media, mice blood and liver extracts, thereby contributing to the understanding of the cellular and physiological metabolic responses to fasting and to oncogene activation, which are important aspects of tumour biology.” Ramón Campos-Olivas Unit Head Technician Clara M. Santiveri ( TS )* *Titulado Superior ( Advanced Degree ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 85 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME | SPECTROSCOPY AND NUCLEAR MAGNETIC RESONANCE UNIT RESEARCH HIGHLIGHTS Our Core Unit incorporates a broad range of instrumentation for the biophysical characterisation of biomolecules and their interactions, including spectrophotometers, a fluorimeter, isothermal titration and differential scanning calorimeters, a circular dichrograph, a multi-angle static light scattering apparatus, and a surface plasmon resonance ( SPR ) instrument. Research groups mostly from, but not limited to, the Structural Biology and Biocomputing Programme have extensively used these technologies throughout 2016. For example, we reported the results of a multidisciplinary intra-Programme collaboration this year ( with the former Computational Biophysics Group ), illustrated in the FIGURE, which combines various NMR and SPR experiments with enhanced sampling molecular dynamics simulations to shed light on the conformational dynamics associated with the binding of Imatinib to the proto-oncogene c-Src. We found that both conformational selection and induced fit play a role in the binding mechanism, reconciling opposing views held in the literature. The Unit hosts a 700 MHz NMR spectrometer, which is well equipped with probes, and a sample changer for running up to 120 samples automatically. This provided the required throughput for screening small molecule protein binders ( together with the CNIO’s Structural Biology and Biocomputing and Experimental Therapeutics -ETP- Programmes ), as well as for metabonomics measurements that were performed in collaboration with the CNIO-Lilly Cell Signalling Therapies Section ( from the ETP ), the Tumour Suppression Group ( from the Molecular Oncology Programme ), as well as the Genes, Development and Disease and the Growth Factors, Nutrients and Cancer Groups ( from the Cancer Cell Biology Programme ). Collectively, with these and previous groups, we implemented sample preparation protocols and developed spectroscopic and analysis technology to characterise the metabolites present in different biological samples, as illustrated by two important publications. s ∞ PUBLICATIONS ∞ Burén S, Gomes AL, Teijeiro A, Fawal MA, Yilmaz M, Tummala KS, Perez M, Rodri- guez-Justo M, Campos-Olivas R, Megías D, Djouder N ( 2016 ). Regulation of OGT by URI in Response to Glucose Confers c-MYC-Dependent Survival Mechanisms. Cancer Cell 30, 290-307. ∞ Fernández FJ, Ardá A, López-Estepa M, Aranda J, Peña-Soler E, Garces F, Round A, Campos-Olivas R, Bruix M, Coll M, Tuñón I, Jiménez-Barbero J, Vega MC ( 2016 ). Mech- anism of sulfur transfer across protein-pro- tein interfaces : The cysteine desulfurase model system. ACS Catal, 6, 3975–3984. ∞ Assi HA, Harkness RW 5th, Martin-Pintado N, Wilds CJ, Campos-Olivas R, Mittermai- er AK, González C, Damha MJ. ( 2016 ). Stabilization of i-motif structures by 2 ’-ß-fluorination of DNA. Nucleic Acids Res 44, 4998-5009. ∞ López-Guadamillas E, Fernández-Marcos PJ, Pantoja C, Muñoz-Martin M, Martínez D, Gómez-López G, Campos-Olivas R, Valverde AM, Serrano M ( 2016 ). p21Cip1 plays a critical role in the physiological adaptation to fasting through activation of PPARa. Sci Rep 6, 34542. ∞ Morando MA, Saladino G, D’Amelio N, Pucheta-Martinez E, Lovera S, Lelli M, López-Méndez B, Marenchino M, Cam- pos-Olivas R, Gervasio FL ( 2016 ). Con- formational Selection and Induced Fit Mechanisms in the Binding of an Anti- cancer Drug to the c-Src Kinase. Sci Rep 6, 24439. ∞ Bayó-Puxan N, Rodríguez-Mias R, Goldflam M, Kotev M, Ciudad S, Hipolito CJ, Varese M, Suga H, Campos-Olivas R, Barril X, Gual- lar V, Teixidó M, García J, Giralt E. ( 2016 ). Combined Use of Oligopeptides, Frag- ment Libraries, and Natural Compounds : A Comprehensive Approach To Sample the Druggability of Vascular Endothelial Growth Factor. ChemMedChem 11, 928-939. ∞ Barbero N, Marenchino M, Campos-Olivas R, Oliaro-Bosso S, Bonandini L, Boskovic J, Viscardi G, Visentin S ( 2016 ). Nano- materials-protein interactions : the case of pristine and functionalized carbon nanotubes and porcine gastric mucin. J Nanopart Res 18, 84. Book Chapter ∞ Santiveri CM, López-Méndez B, Huecas S, Alfonso C, Luque-Ortega, JR, Campos-Ol- ivas R. A biophysical toolkit for molecular interactions. In : eLS. John Wiley & Sons Ltd, Chichester. http ://www.els.net/ In press ( A27015 ). Figure Free energies for local unfolding represented on the backbone structure of the kinase domain of Src in its free form ( left ) as well as their variation upon binding to Imatinib ( right ). The indicated values are coded both in the colour and in the thickness of the backbone coil. They were derived from the rates of exchange of backbone amide protons measured from H/D exchange NMR measurements for all but grey-coloured residues. Red circles indicate a-helices aD and aG ; the regions with increased exposure to the solvent in the bound structure. ANNUAL REPORT 2016 86 VICE-DIRECTION OF BASIC RESEARCH BIOINFORMATICS UNIT OVERVIEW Bioinformatics is a key discipline for understanding the cancer genome and, therefore, essential for the future of cancer therapeutics. Bioinformatics-based approaches have the ability to transform the huge amount of biological data into comprehensive models that provide an in-depth understanding of cancer disease and the complex relationships between genotype and phenotype that are needed to identify cancer driver molecular alterations and new therapeutic targets. Our Unit has several goals : ɗ To provide bioinformatics support with data analysis and interpretation using computational and statistical methods. ɗ To achieve genome analysis in cancer patients ’ data in order to identify new biomarkers and mechanisms of drug response. “ We have developed SATIE, a tool that enables us to predict sequential treatments in cancer. SATIE can propose sensitising treatments, second-line therapies or therapeutic interventions for acquired drug resistance.” ɗ To develop new computational methodologies and bioinformatics tools for cancer research. ɗ To maintain CNIO’s scientific computing facilities and to provide training in bioinformatics tools and methods. Fátima Al-Shahrour Unit Head Post-Doctoral Fellow Hector Tejero SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 87 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME | BIOINFORMATICS UNIT RESEARCH HIGHLIGHTS During 2016, the Bioinformatics Unit ( former Head, David G. Pisano ), and the Translational Bioinformatics Unit headed by Fátima Al-Shahrour from the Clinical Research Programme, were reorganised and merged into one single Bioinformatics Unit ( BU ). BU was established with the aim of providing resources to enable the integration of biological and clinical data, using computational biology approaches, as well as to contribute to research projects in need of bioinformatics support. In 2016, BU published 22 peer-reviewed articles as a result of our ongoing research projects and scientific collaborations with CNIO Research Groups as well as other national and international research institutions. We developed several bioinformatics tools for the analysis of next-generation sequencing data in collaboration with the SING group from the University of Vigo – RubioSeq+ ( Rubio-Camarillo et al., 2017 ), nextpresso ( Graña et al., 2016 ) – and 2 web tools to guide the selection of therapies from genome-wide studies in cancer disease – PanDrugs ( http :// pandrugs.bioinfo.cnio.es ) and SATIE ( http ://satie.bioinfo.cnio. es ). RubioSeq+ has been used in several projects, such as the analysis of the lynx genome ( Abascal et al., 2016 ), and for the whole-exome sequencing analysis of patient-derived xenografts for lung cancer ( Pereira et al., 2016 ). Regarding our scientific collaborations, we helped to unveil mechanisms of cellular reprogramming and senescence ( Mosteiro et al., 2016 ), and also to describe the role of p21 in fasting adaptation ( Lopez-Guadamillas et al., 2016 ) in collaboration with Manuel Serrano’s Group ( CNIO ). Other bioinformatics analyses were performed together with Mariano Barbacid’s Group ( CNIO ) ( Ambrogio et al., 2016 ); these identified DDR1/Notch inhibition as a novel therapy for KRAS-driven lung adenocarcinoma. Finally, within the context of our international collaborations with Harvard associated institutions, we have contributed to the study of leukaemia stem cells in AML ( Puram et al. 2016 ) and the mechanisms of CALR mutations in MPN cells ( Elf et al., 2016 ). s Graduate Student Javier Perales Technicians Ángel Carro ( TS )*, Coral Fustero ( PEJ-L )**, Gonzalo Gómez ( TS )*, Osvaldo Graña ( TS )*, Elena Piñeiro ( TS )*, Miriam Rubio ( TS )*, Kevin Troulé ( PEJ-L )** *Titulado Superior ( Advanced Degree ) **Plan de Empleo Joven-Licenciado ( Youth Employment Plan-Graduate ) ∞ SELECTED PUBLICATIONS * ∞ Mosteiro L, Pantoja C, Alcazar N, Marión RM, Chondronasiou D, Rovira M, Fernandez-Mar- cos PJ, Muñoz-Martin M, Blanco-Aparicio C, Pastor J, Gómez-López G, De Martino A, Blasco MA, Abad M, Serrano M ( 2016 ). Tissue damage and senescence provide critical signals for cellular reprogramming in vivo. Science 354, 6315. pii : aaf4445. ∞ Ambrogio C, Gómez-López G, Falcone M, Vidal A, Nadal E, Crosetto N, Blasco RB, Fernández-Marcos PJ, Sánchez-Céspedes M, Ren X, Wang Z, Ding K, Hidalgo M, Serrano M, Villanueva A, Santamaría D, Barbacid M ( 2016 ). Combined inhibition of DDR1 and Notch signaling is a ther- apeutic strategy for KRAS-driven lung adenocarcinoma. Nat Med 22, 270-277. ∞ Puram RV et al. ( incl. Al-Shahrour F ) ( 2016 ). Core Circadian Clock Genes Regulate Leukemia Stem Cells in AML. Cell 165, 303-316. ∞ Elf S Abdelfattah NS et al. ( incl. Al-Shah- rour F ) ( 2016 ). Mutant Calreticulin Re- quires Both Its Mutant C-terminus and the Thrombopoietin Receptor for Oncogenic Transformation. Cancer Discov 6,368-381. ∞ Montero JJ, López de Silanes I, Graña O, Blasco MA ( 2016 ). Telomeric RNAs are essential to maintain telomeres. Nat Commun 17, 12534. ∞ Pérez-Guijarro E, Karras P, Cifdaloz M, Martínez-Herranz R, Cañón E, Graña O, Horcajada-Reales C, Alonso-Curbelo D, Calvo TG, Gómez-López G, Bellora N, Riveiro-Falkenbach E, Ortiz-Romero PL, Rodríguez-Peralto JL, Maestre L, Ronca- dor G, de Agustín Asensio JC, Goding CR, Eyras E, Megías D, Méndez R, Soengas MS ( 2016 ). Lineage-specific roles of the cyto- plasmic polyadenylation factor CPEB4 in the regulation of melanoma drivers. Nat Commun 7, 13418. ∞ Abascal F et al. ( incl. Tress ML, Valencia A ) ( 2016 ). Extreme genomic erosion after recurrent demographic bottlenecks in the highly endangered Iberian lynx. Genome Biol 17, 251. *please see BU’s web site for a list of all publications. Figure SATIE-Sequential Antitumour Treatment Inference and Enrichment tool. http ://satie.bioinfo. cnio.es ANNUAL REPORT 2016 88 VICE-DIRECTION OF BASIC RESEARCH NATIONAL BIOINFORMATICS INSTITUTE UNIT OVERVIEW The Spanish National Bioinformatics Institute ( Instituto Nacional de Bioinformática, INB ) is a component of the National Infrastructure of Biomolecular and Bioinformatics Resources Platform ( Plataforma en Red de Recursos Biomoleculares y Bioinformáticos, PRB2 ) of the Spanish National Institute of Health Carlos III ( Instituto de Salud Carlos III, ISCIII ). The INB is the Spanish Node of ELIXIR, the permanent European Infrastructure for Life Sciences. The INB is composed of 10 working nodes distributed across 9 different research centres. The INB Unit at the CNIO undertakes the coordination of the institute. As the coordination node, the goals of the INB Unit are to : ɗ Coordinate the Spanish participation in ELIXIR. Promote the implementation and adoption of ELIXIR guidelines among the Spanish bioinformatics community. “ The INB Unit has actively participated in the management of data portals for big research consortia like BLUEPRINT, ICGC, and PanCancer, aiming to understand the genetic bases of cancer.” ɗ Design, promote and ensure the execution of the INB’s scientific-technical and training programmes, undertaken with the support of all nodes. ɗ Promote the collaboration between INB nodes and third parties, including research consortia, other research infrastructures, small and medium enterprises ( SMEs ), and the industry. Salvador J. Capella Gutierrez ( since June ) Unit Head Technicians Andrés Cañada ( TS )*, José M. Fernández ( TS )*, José M. Rodríguez ( TS )* *Titulado Superior ( Advanced Degree ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 89 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME | NATIONAL BIOINFORMATICS INSTITUTE UNIT RESEARCH HIGHLIGHTS Despite this Unit’s coordination role, the INB actively participates in different Work Packages ( WP ) of the ELIXIR-EXCELERATE programme. It is also involved in other major projects such as RD-Connect, BLUEPRINT and eTOX. The Unit’s contribution can be divided into three main areas : Data resources and Bio-computing The storage and processing of data have become fundamental tasks within almost all of the current research projects. Through a collaboration model, the Unit participates in several research projects studying the data requirements and developing solutions to store and process the data. An example of this is the BLUEPRINT data-portal ( http ://dcc.blueprint-epigenome.eu ). BLUEPRINT is a high impact FP7 project aimed at producing epigenomes of haemopoetic cell lines from healthy and non- healthy human donors. In the current version, the data portal provides an epigenomic analysis, obtained from 1,019 samples, to the scientific community. Their associated epigenomes are characterised by : gene and transcript expression ( from RNA-Seq experiments ), hyper and hypo methylated regions ( derived from WGBS experiments ), chromatin accessibility ( DNAse-Seq ), and 7 Histone marks binding activity ( ChIP- Seq ). Recently, a scientific article was published illustrating the possibilities offered by this portal ( Fernandez JM et al, 2016 ). Infrastructure development Within the infrastructure development aspect, there is a clear focus on developing a text-mining infrastructure for the processing of biomedical texts. The LiMTox system ( http :// limtox.bioinfo.cnio.es ) is the first text mining approach designed to extract associations between compounds and a particular toxicological end point at various levels of granularity and evidence types, all inspired by the content of toxicology reports. During this time, a second end-point ( http ://melanomamine. bioinfo.cipf.es ) has been built using the same system and focusing on the study of different aspects of melanomas. End users applications and services The Unit actively contributes to the creation of an integrated platform that connects databases, registries, biobanks and clinical bioinformatics for research on rare diseases. RD-Connect ( http :// rd-connect.eu ) allows researchers and clinicians to explore the possible genetic causes of these diseases by combining genomic data with phenotypic information from patients across Europe, in a clear attempt to have enough statistical power to support findings. The INB unit has also developed APPRIS ( http ://appris. bioinfo.cnio.es ) and keeps improving it in order to annotate genes and identify the principal isoform of every single gene. Currently, the GENCODE consortium uses APPRIS to annotate the principal human and mouse isoforms. s ∞ PUBLICATIONS ∞ Chen L et al. ( incl. Fernandez JM, Rico D, Valencia A ) ( 2016 ). Genetic Drivers of Epigenetic and Transcriptional Variation in Human Immune Cells. Cell 167, 1398–1414. ∞ Abascal F et al. ( incl. Capella-Gutierrez S, Valencia A ) ( 2016 ). Extreme genomic erosion after recurrent demographic bot- tlenecks in the highly endangered Iberian lynx. Genome Biology 17, 251. ∞ Ecker S et al. ( incl. Fernandez JM, Rico D, Valencia A ) ( 2016 ). Genome-wide Analysis of Differential Transcriptional and Epigenetic Variability Across Human Immune Cell Types. BioRxiv, 83246. ∞ Fernández JM, de la Torre V, Richardson D, Royo R, Puiggròs M, Moncunill V, Fragkogi- anni S, Clarke L ; BLUEPRINT Consortium., Flicek P, Rico D, Torrents D, Carrillo de Santa Pau E, Valencia A ( 2016 ). The BLUEPRINT Data Analysis Portal. Cell Syst 3,491-495. ∞ PUBLICATIONS AT OTHER INSTITUTIONS ∞ Altenhoff, AM et al. ( incl. Capella-Gutierrez S ) ( 2016 ). Standardized benchmarking in the quest for orthologs. Nat Methods 13, 425–430. ∞ Vlasova A et al. ( incl. Capella-Gutierrez S ) ( 2016 ). Genome and Transcriptome Analysis of the Mesoamerican Common Bean and the Role of Gene Duplications in Establishing Tissue and Temporal Specialization of Genes. Genome Biol 17, 32. ∞ Sebé-Pedros A, Peña MI, Capella-Gut- ierrez S, Antó M, Gabaldón T, Ruíz-Trillo I & Sabidó E. ( 2016 ). High-throughput proteomics reveals the unicellular roots of animal phosphosignaling and cell dif- ferentiation. Dev Cell 39, 186-197. Figure A collage of the different services developed by the INB Unit. Top left shows the experimental information available at BLUEPRINT. Top right shows the information available at the APPRIS home-page. Bottom panels feature information found for Ibuprofen using the LimTox system. ANNUAL REPORT 2016 90 VICE-DIRECTION OF BASIC RESEARCH ELECTRON MICROSCOPY UNIT OVERVIEW The Electron Microscopy ( EM ) Unit is a central core facility and a research laboratory that provides CNIO researchers, as well as the broader research community, with access to Transmission Electron Microscopy and also provides expertise in EM image analysis. As a core facility, we offer standard specimen preparation techniques for proteins, protein complexes and vesicles, data collection and data processing tailored to the particular needs of the users. We also offer training for regular users on the use of equipment, as well as guidance regarding specimen preparation. “ We have used single-particle electron microscopy to elucidate the molecular architecture of full-length TRF1 and to demonstrate how it assists its interaction with other proteins and telomeric DNA.” Jasminka Boskovic Unit Head Technicians Alberto Buscató ( until July ) ( PEJ-L )*, Carlos Rodríguez ( since September ) ( PEJ-L )* *Plan de Empleo Joven-Licenciado ( Youth Employment Plan-Graduate ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 91 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME | ELECTRON MICROSCOPY UNIT RESEARCH HIGHLIGHTS The Electron Microscopy Unit is a research facility that supports biological scientific projects ranging from the cellular to the macromolecular level. The EM Unit performs sample preparation protocols, negative staining, cryo-EM, and data collection methods, as well as 2D and 3D data processing. In collaboration with CNIO’s Telomeres and Telomerase Group ( Molecular Oncology Programme ) and the Crystallography and Protein Engineering Unit ( Structural Biology and Biocomputing Programme ), we used the single-particle electron microscopy technique to obtain the first low resolution structures of full- length TRF1 dimer ( shelterin component ) and its structure in complex with telomeric DNA. We contributed to the understanding of the molecular mechanism that protects the ends of chromosomes : our results demonstrate that full-length TRF1 presents a molecular architecture that assists its interaction with telomeric DNA and at the same time makes TRFH domains accessible to other TRF1 binding partners. Furthermore, our studies suggest hypothetical models on how other proteins such as TIN2 and tankyrase contribute to regulate TRF1 function. In collaboration with Iván Ventoso, from the Centro de Biología Molecular ‘ Severo Ochoa ’ ( CSIC-UAM ) and the Departamento de Biología Molecular, Universidad Autónoma de Madrid ( UAM ), the EM Unit participated in the novel findings that illustrate how viral mRNA is threaded into the 40S subunit during the scanning process. Based on structural and functional data, we generated new insights into the scanning process, describing how a stem-loop in the proximal region of viral mRNA can promote a Eukaryotic Initiation Factor 2 ( eIF2 )-less translation initiation by trapping in RNA extensions of the ribosomal 40S subunit. We continued our collaboration with the CNIO Cell Signalling and Adhesion Group ( Structural Biology and Biocomputing Programme ) on PI( 4,5 )P2-mediated induction of Focal Adhesion Kinase ( FAK ) clustering at the cell membrane, applying 2D electron crystallography. s ∞ PUBLICATIONS ∞ Toribio R, Díaz-López I, Boskovic J, Vento- so I ( 2016 ). An RNA trapping mechanism in Alphavirus mRNA promotes ribosome stalling and translation initiation. Nucleic Acids Res 44, 4368-4380. ∞ Boskovic J, Bragado-Nilsson E, Saligram Prabhakar B, Yefimenko I, Martínez-Gago J, Muñoz S, Méndez J, Montoya G ( 2016 ). Molecular architecture of the recombi- nant human MCM2-7 helicase in complex with nucleotides and DNA. Cell Cycle 15, 2431-2440. ∞ Boskovic J, Martinez-Gago J, Mendez-Per- tuz M, Buscato A, Martinez-Torrecuadrada JL, Blasco MA ( 2016 ). Molecular Archi- tecture of Full Length TRF1 Favors its Interaction with DNA. J Biol Chem 291, 21829-21835. ∞ Barbero N, Marenchino M, Campos-Olivas R, Oliaro-Bosso S, Bonandini L, Boskovic J, Viscardi G, Visentin S ( 2016 ). Nano- materials-protein interactions : the case of pristine and functionalized carbon nanotubes and porcine gastric mucin. J Nanopart Res 18, 84. Figure TRF1 structure (  A   ) and model for TRF1 DNA binding ( B ) and release ( C ). The interaction with TIN2 stabilises the complex with DNA through direct interaction of TIN2 with DNA. Tankyrase 1 engages the TRF1 dimer on two opposite sides of the molecule, introducing the PARylation and the release of TRF1. ANNUAL REPORT 2016 92 VICE-DIRECTION OF BASIC RESEARCH CRYSTALLOGRAPHY AND PROTEIN ENGINEERING UNIT OVERVIEW Nowadays, knowledge of the three-dimensional ( 3D ) structure of a protein is critical in order to gain a full understanding of its function. The structures of proteins, alone or in complex with other biological partners, reveal functional networks thereby providing a better understanding of the behaviour of the cell’s molecular machinery. This implies knowing how proteins move, detecting their interacting partners, and comprehending the changes that they undergo. Close images facilitated by 3D structures provide the possibility of introducing rationally designed mutants that alter their affinity and specificity towards interacting molecules, aiding in the recognition of the physicochemical mechanisms that govern their function. This is why structural data has become crucial in guiding the drug design process, and the results have proven to be relevant for the development of novel therapies. To achieve this goal, the Crystallography and Protein Engineering Unit provides services at different levels in order to meet the demands of research groups at the CNIO and outside our institute. At the structural determination level, the Unit offers state-of-the- art, high-throughput protein crystallisation screening facilities that include sophisticated equipment for the identification of protein crystals, as well as a full-service offering of X-ray crystallography and small-angle x-ray ( SAXS ) analyses. As an academic Unit, we have access to high-tech European infrastructures such as the synchrotron light sources. At the protein production level, we have at our disposal a wide array of instrumentation and technical support for the design and purification of soluble recombinant proteins required in large amounts up to milligram quantities, for structural, biophysical or biochemical characterisation, and also for antibody production. Inés Muñoz Unit Head Staff Scientist Jorge L. Martínez SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 93 STRUCTURAL BIOLOGY AND BIOCOMPUTING PROGRAMME | CRYSTALLOGRAPHY AND PROTEIN ENGINEERING UNIT RESEARCH HIGHLIGHTS This has been a year of growth and exciting changes for this ‘ new ’ Crystallography and Protein Engineering Unit. It is the result of the inclusion of CNIO’s protein production facility, previously integrated in the Proteomics Unit ( Biotechnology Programme ), in the Crystallography Unit. The Unit continues to be shared between the Structural Biology and Biocomputing Programme and the Experimental Therapeutics Programme. Throughout 2016, we have worked closely with the Experimental Therapeutics Programme on several projects, some of them also in collaboration with other CNIO Groups. The scaling up of the production of proteins, like full-length human MASTL, has permitted a wide range of biochemical experiments to take place. Other projects were directly focused on structural characterisation by x-ray crystallography in support of drug discovery, as in the case of the human proteins HASPIN and CDK8/CyclinC complex where we obtained several crystal structures of the protein-ligand complexes ( FIGURE ). Especially relevant was our continuous work on the production of proteins for the generation of antibodies by the CNIO Monoclonal Antibody Unit ( Biotechnology Programme ). During 2016, this smooth collaboration has led to the production of several proteins involved in cancer such as CDC25A, IDO1, TDO2, IL11, PDL1, PDL2 or NOMO1. The Unit also undertakes several collaborations with different CNIO groups. It is noteworthy to mention the collaborations established with CNIO’s Telomeres and Telomerase Group, the Gastrointestinal Cancer Clinical Research Unit, the Epithelial Carcinogenesis Group and the Structural Computational Biology Group. Additionally, the Unit maintains external collaborations with groups at the Physical Chemistry Department ( University of Granada ), the Environmental Biology Department ( CIB-CSIC ), the Pharmacology and Therapeutics Department ( Roswell Park Cancer Institute, USA ), the Department of Biomedicine ( University of Bergen, Norway ), and the Department of Molecular Engineering ( Åarhus University, Denmark ). Finally, the Unit has continued the study of the role of ephrinB2 in different pathologies. This was done by blocking its activity with specific recombinant antibodies generated by us, in collaboration with groups from the MRC Clinical Sciences Centre ( UK ) and the NCI Center for Cancer Research ( USA ). s Student in Practice Silvia L. Gomes Technicians Daniel Calvo ( since February ) ( PEJ-L )*, Jaime Martínez ( until April ) ( TS )**, Álvaro Otero ( PEJ-L )*, Alicia Virseda ( PEJ-L )* *Plan de Empleo Joven-Licenciado ( Youth Employment Plan-Graduate ) **Titulado Superior ( Advanced Degree ) ∞ PUBLICATIONS ∞ Krusche B, Ottone C, Clements MP, John- stone ER, Goetsch K, Lieven H, Mota SG, Singh P, Khadayate S, Ashraf A, Davies T, Pollard SM, De Paola V, Roncaroli F, Martin- ez-Torrecuadrada J, Bertone P, Parrinello S ( 2016 ). EphrinB2 drives perivascular invasion and proliferation of glioblastoma stem-like cells. Elife pii, e14845. ∞ Pucheta-Martínez E, Saladino G, Morando MA, Martinez-Torrecuadrada J, Lelli M, Sutto L, D’Amelio N, Gervasio FL ( 2016 ). An Allosteric Cross-Talk Between the Ac- tivation Loop and the ATP Binding Site Regulates the Activation of Src Kinase. Sci Rep 6, 24235. ∞ Pucheta-Martinez E, D’Amelio N, Lelli M, Martinez-Torrecuadrada JL, Sudol M, Saladino G, Gervasio FL ( 2016 ). Chang- es in the folding landscape of the WW domain provide a molecular mechanism for an inherited genetic syndrome. Sci Rep 6, 30293. ∞ Boskovic J, Martinez-Gago J, Mendez-Pertuz M, Buscato A, Martinez-Torrecuadrada JL, Blasco MA ( 2016 ). Molecular Architecture of Full Length TRF1 Favors its Interaction with DNA. J Biol Chem 291, 21829-21835. ∞ Fuchs JE, Muñoz IG, Timson DJ, Pey AL ( 2016 ). Experimental and computational evidence on conformational fluctuations as a source of catalytic defects in genetic diseases. RSC Adv 6, 58604-58612. ∞ de Lucas AG, Schuhmacher AJ, Oteo M, Romero E, Cámara JA, de Martino A, Arroyo AG, Morcillo MÁ, Squatrito M, Martinez-Torrecuadrada JL, Mulero F ( 2016 ). Targeting MT1-MMP as an im- munoPET-based strategy for imaging gliomas. PLoS One 11, e0158634. ∞ Flydal MI, Alcorlo M, Skjaerven S, Muñoz IG, Teigen K, Hermoso JA, Martinez A ( 2016 ). Structural and dynamics studies of human phenylalanine hydroxilase, a highly regulated allosteric enzime. Acta Crystallogr A A72, S239. Figure Close views of the active sites from the x-ray structures of HASPIN ( A ) and CDK8/CyclinC ( B ). The coloured omit maps correspond to the electron density of the bound compounds ( synthesised in CNIO’s Medicinal Chemistry Section ). ( C ) SAXS ab initio shape reconstructions of wild type UDP- galactose 4’-epimerase ( GALE ) and its mutant G90E, superimposed on the crystal structure. The data explains the drastic conformational changes that reduce NAD+ binding affinity in the mutant, causing type III galactosemia. This work was done in collaboration with the Physical Chemistry Department ( University of Granada, Spain ). ANNUAL REPORT 2016 94 Vice-Direction of Translational Research SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 95 Human Cancer Genetics Programme 98 Human Genetics Group 100 Hereditary Endocrine Cancer Group 104 Genetic and Molecular Epidemiology Group 108 Familial Cancer Clinical Unit 112 Molecular Cytogenetics and Genome Editing Unit 114 Human Genotyping-CEGEN Unit 116 Clinical Research Programme 118 Gastrointestinal Cancer Clinical Research Unit 120 Breast Cancer Junior Clinical Research Unit 122 Prostate Cancer Junior Clinical Research Unit 124 Molecular Diagnostics Unit 126 H12O-CNIO Haematological Malignancies Clinical Research Unit 128 H12O-CNIO Lung Cancer Clinical Research Unit 130 Biobank 132 ANNUAL REPORT 2016 96 ÓSCAR FERNÁNDEZ-CAPETILLO Vice-Director of Translational Research SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 97 “ With our translational research efforts, at the CNIO we are trying to reduce the gap between research laboratories and cancer patients.” The ultimate goal of the CNIO is to contribute to the global effort in the fight against cancer. This involves the work of many different laboratories and individuals, who collectively help to achieve this mission. During 2016, the CNIO maintained an important focus on translational research, placing significant importance on those research aspects that are in close proximity to the patients. These included the identification of novel mutations and altered pathways in samples from cancer patients that could perhaps be used to guide novel treatments. It is noteworthy to mention that a lot of this work was done under the framework of large international Consortia in which we participated, again underscoring CNIO’s active contribution towards the worldwide effort in cancer research. One example of our key role in collaborative efforts is the important function that the CNIO Biobank carries out in coordinating the Spanish National Biobank Network, which manages patient samples from 52 institutions across Spain. Finally, and in addition to our work in the discovery of molecular alterations that are present in cancer patients, the Clinical Research Programme tries to capitalise on these discoveries ( as well as others from all around the world ) by bringing them to the clinic. Besides from their efforts in trying to implement new clinical trials, the research from our clinical groups has revealed new insights that can be used to benefit cancer patients, such as novel strategies directed towards overcoming resistance to antiangiogenic agents. Finally, through our collaborative agreement with Hospital 12 de Octubre, two Clinical Research Units established by investigators from that hospital are also housed at the CNIO, in an effort to further strengthen our links with neighbouring hospitals. ANNUAL REPORT 2016 98 VICE-DIRECTION OF TRANSLATIONAL RESEARCH HUMAN CANCER GENETICS PROGRAMME JAVIER BENÍTEZ Director SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 99 HUMAN CANCER GENETICS PROGRAMME | The Human Cancer Genetics Programme is currently composed of three Research Groups : Human Genetics, Endocrine Cancer, and Genetic and Molecular Epidemiology Groups ; and three Units : Human Genotyping-CEGEN and Molecular Cytogenetics and Genome Editing Units ; and the Familial Cancer Clinical Unit. In addition, there is a Familial Cancer Consultancy for the evaluation of families with cancer and the provision of genetic counselling. The Consultancy is located in the Hospital Universitario de Fuenlabrada and works in close collaboration with the Oncology Service at that Hospital. The number of consultancy days and amount of families attended have increased since we set it up five years ago. Currently, we are attending to around 350 families/ year. This increase of families has resulted in a higher number of genetic and genomic diagnosis studies, which have been made possible thanks to the incorporation of a massive sequencing platform. This platform has been operational over the past year. The Programme’s core goals are centred on research, training and diagnosis. The genetic and cytogenetic study of tumours, genome editing, genetic interactions, data integration, the search for diagnostic and prognostic markers, the discovery of novel cancer-related genes and environmental factors that confer cancer susceptibility, are our main research priorities. A further complementary area of work is the application of Pharmacogenetics and Pharmacogenomics to identify genes that modify drug response. This research line focuses on a wide variety of tumours, taking advantage of the high-throughput genotyping technologies provided by the Genotyping Unit. The Programme collaborates closely with the clinical community, not only to foster cooperation in genetic diagnosis but also to promote in training and education. During this year, the Programme’s groups have hosted 8 residents from different Spanish hospitals for 3-month periods. We also offer professionals from different international research centres the opportunity to join us, either as visitors or for longer training visits consisting of short-term stays of 1-3 months ( a total of 3 international visitors from Latin America were hosted in 2016 ). In terms of education, since the beginning of 2016, 1 foreign and 3 national Erasmus Master’s students and 9 national and 2 international PhD students have worked on their research projects, 1 of whom has already successfully defended her thesis. We participate in many international and national consortia. This enables us to apply for international projects, hold international meetings and publish in the best journals. Likewise, a good collaboration with other CNIO Groups and Units is one of our main characteristics, allowing us to benefit from the internal exchange connecting people, techniques, technology and knowledge. Milestones and major achievements of the Programme in 2016 include : ɗ The co-organisation of the 6th Familial Cancer Conference in collaboration with the European School of Oncology and Nature Reviews Clinical Oncology. ɗ Mercedes Robledo, Head of the Hereditary Endocrine Cancer Group, was awarded the International Medal bestowed by the Society for Endocrinology. ɗ Núria Malats, Head of the Genetic and Molecular Epidemiology Group, was elected Chair of the EUPancreas COST Action. ɗ Co-coordinating the BDebate on the Human Microbiome. CaixaForum, Barcelona, 29 June - 1 July 2016. ɗ The co-direction of the CNIO Canceromatics III - Tumor Heterogeneity Conference, November 2016. “ The programme continues with carrying out its translational work, connecting clinicians with scientists, studying human cancers and helping and advising professionals about the new genetic results generated by novel technologies.” ANNUAL REPORT 2016 100 VICE-DIRECTION OF TRANSLATIONAL RESEARCH HUMAN GENETICS GROUP Javier Benítez Group Leader Staff Scientists M. José García, Ana Osorio SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 101 HUMAN CANCER GENETICS PROGRAMME | HUMAN GENETICS GROUP OVERVIEW The Human Genetics Group is working on the study of human cancer from a genetic, cytogenetic and epidemiologic point of view. We want to understand why the inherited susceptibility to cancer doesn’t follow a mathematical model in people, but rather ‘ an apparently random model ’, and why there are families with a large number of members suffering from cancer. For these studies we work with individuals, families and the affected and normal population, trying to perform a correct diagnosis with known genes as well as looking for new genes that could explain cancer susceptibility in specific families. Our main objective is to work with every family by raising their awareness in regards to their own risk of developing cancer and how to prevent it. To this primary level, we have to add a secondary level of prevention, which will facilitate an important risk reduction in the population, through the development of non-invasive and non-genetic but yet extremely effective measures. “ During 2016, we showed how inhibitors, other than PARPi, could be used in patients with BRCA mutations. We started working on the identification of new treatments for cardiac tumours based on transcriptome analysis, and finally, we are also exploring a polygenic inheritance model in families with testicular cancer that is based on more than 25 identified genes associated with this disease.” RESEARCH HIGHLIGHTS Breast cancer : PARP1 and OGG1 inhibitors in BRCA1 mutation carriers We have demonstrated that certain missense mutations in BRCA1 seem to make cells more sensitive to Poly ( ADP-ribose ) Polymerase ( PARP ) inhibitors than those mutations that give rise to the absence of the protein ( frameshift mutations ) ( T. Valclová, Hum Mol Genet 2016 ). We are currently investigating the mechanisms underlying these differences with the aim of identifying new markers of sensitivity or resistance to these agents. In parallel, we recently showed that the Single Nucleotide Polymorphism ( SNP ) rs2304277, located in the 3 ’ untranslated region ( UTR ) of the OGG1 DNA glycosylase gene of the Base Excision Repair pathway ( BER ), modified cancer risk in patients harbouring mutations in BRCA1 ( Osorio A. et al., Plos Genetics, 2014 ). We have identified that the SNP is associated with a constitutive hOGG1 transcriptional downregulation, which leads to a high genome and telomere instability in those patients harbouring BRCA1 and BRCA2 mutations, thereby explaining the contribution of this polymorphism to cancer risk. This association is most likely explained by a synthetic lethal/sick interaction between these 2 genetic events. ( Benitez-Buelga C. et al., Oncotarget, 2016 ). In order to take an in-depth look at the biological link between BER and the homologous recombination ( HR ) DNA repair pathway, we tested the pharmacological inhibition of OGG1 in a set of BRCA1 and BRCA2 deficient cancer cell lines. We found that OGG1 inhibition is effective, leading to 1 ) an accumulation of telomere oxidation ( genomic instability ), and 2 ) an alteration in the normal proliferation of BRCA1 deficient cell lines, pointing to a synthetic lethal interaction between OGG1 and BRCA1 ( FIGURE 1 ). Familial cancer exome project This project started several years ago with the objective of identifying new high susceptibility genes that explain families Post-Doctoral Fellows Oriol Calvete, Javier Gayarre ( until July ) Graduate Students Juan Miguel Baquero, Paloma Martín ( since March ), Beatriz Paumard, Alejandra Tavera Technicians Alicia Barroso, Carlos Benítez-Buelga ( TS )*, M. Victoria Fernández ( TS )* *Titulado Superior ( Advanced Degree ) ANNUAL REPORT 2016 102 VICE-DIRECTION OF TRANSLATIONAL RESEARCH with rare tumours as well as deciphering the genetic heterogeneity present in some of them : ɗ In 2015, we identified ATP4a as being responsible for families with gastric neuroendocrine tumours. We are currently searching for new genes in two families that cannot be explained by mutations in ATP4a. ɗ A second gene, POT1, which was published in 2015 as being associated with familial cardiac angiosarcoma, also explains some families with Li Fraumeni-like syndrome. Analysis of tumour samples by Next Generation Sequencing ( NGS ) has shown an over-representation of the angiogenic pathway, which may be useful in clinical trials. In collaboration with M. Blasco’s Telomeres and Telomerase Group, we are generating a knock-in mouse with the aim of recapitulating the disease, as well as enabling us to work with antiangiogenic drugs. ɗ We are currently investigating a large family with meningiomas across 3 generations. Analysis of the data generated using ∞ PUBLICATIONS ∞ Dunning AM et al. ( incl. González-Neira A, Osorio A, Benítez J ) ( 2016 ). Breast can- cer risk variants at 6q25 display different phenotype associations and regulate ESR1, RMND1 and CCDC170. Nat Genet 48, 374-386. ∞ Kar SP et al. ( incl. Benítez J ) ( 2016 ). Genome-wide meta-analyses of breast, ovarian, and prostate cancer association studies identify multiple new susceptibility loci shared by at least two cancer yypes. Cancer Discovery 6, 1052-1057. ∞ Couch FJ et al. ( incl. González-Neira A, Osorio A, Benítez J ) ( 2016 ). Identification of four novel susceptibility loci for oes- trogen receptor negative breast cancer. Nat Commun 7, 11375. ∞ Lawrenson K et al. ( incl. Benítez J,Osorio A ) ( 2016 ). Functional mechanisms under- lying pleiotropic risk alleles at the 19p13.1 breast-ovarian cancer susceptibility locus. Nat Commun 7, 12675. ∞ Ghoussaini M et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Evidence that the 5p12 Variant rs10941679 Confers Susceptibili- ty to Estrogen-Receptor-Positive Breast Cancer through FGF10 and MRPS30 Reg- ulation. Am J Hum Genet 99, 903-911. ∞ Wang Y et al. ( incl. Benítez J ) ( 2016 ). The BRCA1-?11q Alternative Splice Isoform By- passes Germline Mutations and Promotes Therapeutic Resistance to PARP Inhibition and Cisplatin. Cancer Res 76, 2778-2790. ∞ Wyszynski A et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). An intergenic risk locus containing an enhancer deletion in 2q35 modulates breast cancer risk by deregulating IGFBP5 expression. Hum Mol Genet 25, 3863-3876. ∞ Benitez-Buelga C, Vaclová T, Ferreira S, Uri- oste M, Inglada-Perez L, Soberón N, Blasco MA, Osorio A, Benítez J ( 2016 ). Molecular insights into the OGG1 gene, a cancer risk modifier in BRCA1 and BRCA2 mutations carriers. Oncotarget 7, 25815-25825. ∞ Hamdi Y et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Association of breast cancer risk with genetic variants showing differential allelic expression : Identification of a novel breast cancer susceptibility locus at 4q21. Oncotarget 7, 80140-80163. ∞ Matamala N, Vargas MT, González-Cámpo- ra R, Arias JI, Menéndez P, Andrés-León E, Yanowsky K, Llaneza-Folgueras A, Miñam- bres R, Martínez-Delgado B, Benítez J ( 2016 ). MicroRNA deregulation in triple negative breast cancer reveals a role of miR-498 in regulating BRCA1 expression. Oncotarget 7, 20068-20079. ∞ Easton DF et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). No evidence that protein truncating variants in BRIP1 are associated with breast cancer risk : im- plications for gene panel testing. J Med Genet 53, 298-308. ∞ Southey MC et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). PALB2, CHEK2 and ATM rare variants and cancer risk : data from COGS. J Med Genet 53, 800-811. ∞ Abubakar M et al. ( incl. Benítez J ) ( 2016 ). Prognostic value of automated KI67 scor- ing in breast cancer : a centralised eval- uation of 8088 patients from 10 study groups. Breast Cancer Res 18, 104. ∞ Petridis C et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Genetic predisposition to ductal carcinoma in situ of the breast. Breast Cancer Res 18, 22. ∞ Rebbeck TR et al. ( incl. Benítez J ) ( 2016 ). Inheritance of deleterious mutations at both BRCA1 and BRCA2 in an international sample of 32,295 women. Breast Cancer Res 18, 112. ∞ Silvestri V et al. ( incl. Osorio A, Benítez J ) ( 2016 ). Male breast cancer in BRCA1 and BRCA2 mutation carriers : pathology Figure 1 Different effect of OGG1 inhibitors in BRCA1 vs control cell lines. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 103 HUMAN CANCER GENETICS PROGRAMME | HUMAN GENETICS GROUP bioinformatics tools has shown the existence of 2 candidate genes that could be responsible for this familial tumour. We are starting functional studies and are recruiting more families. ɗ Ovarian cancer families are rare and are usually associated with breast cancer. We sequenced the exomes of 9 patients from 5 families and identified 33 rare variants in 28 genes potentially implicated in ovarian cancer risk. By conducting a case control association study we narrowed down the number of candidate missense variants to 10. These, together with 5 high-impact variants ( protein truncating or splicing variants ), will be evaluated in a larger international case control study to finally define their role in ovarian cancer susceptibility. In parallel, we selected a non-described RAD51C missense variant among the identified candidates, and through functional characterisation we were able to determine its pathogenicity and its probable involvement in ovarian cancer risk in one of the families. This finding not only has implications for genetic counselling but also for the potential treatment of affected carriers with PARPi. ɗ Breast cancer. We have performed whole-exome sequencing ( WES ) in 3 BRCAX families ( familial breast cancer families negative for mutations in BRCA1/2 ). One of the families was found to harbour a deleterious mutation in the known breast cancer susceptibility gene ATM. A complete screening of this gene in a set of 400 Spanish breast cancer families showed a prevalence of almost 2% of mutations in ATM, higher than that reported in other populations ( Tavera-Tapia et al., BCRT 2016 ). In another family, we found an excellent candidate breast cancer gene that is currently being screened by targeted NGS in a series of 700 BRCAX families and 700 controls. The third family is still under analysis. ɗ Male breast cancer. We performed WES in a male breast cancer family with an apparently recessive model of inheritance. We have found 7 candidate variants that are currently being validated in a series of 1000 male breast cancer cases and 1000 controls ; this is undertaken in collaboration with Nick Orr’s lab at the Institute of Cancer Research in London. ɗ Testicular cancer. Testicular cancer follows a polygenic model of inheritance. We have studied, by NGS, 35 families with 2 or 3 first degree relatives affected by the disease. The results have been classified according to different inheritance models ; different methods of analysis have been conducted in order to select some candidate genes ( FIGURE 2 ). The candidate variants are currently being genotyped in a set of more than 500 sporadic testicular cancer cases and 500 controls in order to know how many of them could be considered as candidates to be associated with the disease. s data from the Consortium of Investigators of Modifiers of BRCA1/2. Breast Cancer Res 18, 15. ∞ Zeng C et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Identification of inde- pendent association signals and putative functional variants for breast cancer risk through fine-scale mapping of the 12p11 locus. Breast Cancer Res 18, 64. ∞ Darabi H et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Fine scale mapping of the 17q22 breast cancer locus using dense SNPs, genotyped within the Collaborative Oncological Gene-Environment Study ( COGs ). Sci Rep 7, 32512. ∞ Liu J et al. ( incl. Benítez J, González-Neira A ) ( 2016 ). rs2735383, located at a microR- NA binding site in the 3 ’UTR of NBS1, is not associated with breast cancer risk. Sci Rep 6, 36874. ∞ Shi J et al. ( incl. Benítez J ) ( 2016 ). Fine- scale mapping of 8q24 locus identifies multiple independent risk variants for breast cancer. Int J Cancer 139, 1303-1317. ∞ Calvete O, Varro A, Pritchard DM, Barroso A, Oteo M, Morcillo MÁ, Vargiu P, Dodd S, Garcia M, Reyes J, Ortega S, Benítez J ( 2016 ). A knockin mouse model for human ATP4aR703C mutation identified in familial gastric neuroendocrine tumors recapitulates the premalignant condition of the human disease and suggests new therapeutic strat- egies. Dis Model Mech 9, 975-984. ∞ Lei J et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Genetic variation in the immu- nosuppression pathway genes and breast cancer susceptibility : a pooled analysis of 42,510 cases and 40,577 controls from the Breast Cancer Association Consortium. Hum Genet 135, 137-154. ∞ Horne HN et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Fine-Mapping of the 1p11.2 Breast Cancer Susceptibility Locus. PLoS One 11, e0160316. ∞ Pelttari LM et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). RAD51B in Familial Breast Cancer. PLoS One 11, e0153788. ∞ Vigorito E et al. ( incl. Osorio A, Benítez J ) ( 2016 ). Fine-Scale Mapping at 9p22.2 Iden- tifies Candidate Causal Variants That Modify Ovarian Cancer Risk in BRCA1 and BRCA2 Mutation Carriers. PLoS One 1, e015880. ∞ Zhao Z et al. ( incl. Osorio A, Benítez J ) ( 2016 ). Association of genetic susceptibil- ity variants for type 2 diabetes with breast cancer risk in women of European ances- try. Cancer Causes Control 27, 679-693. ∞ Fossmark R, Calvete O, Mjønes P, Benítez J, Waldum HL ( 2016 ). ECL-cell carcinoids and carcinoma in patients homozygous for an inactivating mutation in the gastric H+ K+ ATPase alpha subunit. APMIS 124, 561-566. ∞ Abubakar M et al. ( incl. Benítez J ) ( 2016.) High-throughput automated scoring of Ki67 in breast cancer tissue microarrays from the Breast Cancer Association Con- sortium. J Pathol Clin Res 2, 138-153. ∞ Gayarre J et al. ( incl. Benítez J ) ( 2016 ). The NER-related gene GTF2H5 predicts survival in high-grade serous ovarian cancer patients. J Gynecol Oncol 27, E7. ∞ Hellner K et al. ( incl. Benítez J ) ( 2016 ). Premalignant SOX2 overexpression in the fallopian tubes of ovarian cancer patients : Discovery and validation studies. EBiomed 10, 137-149. ∞ AWARDS AND RECOGNITION ∞ Ana Osorio has been awarded the madri+d Science Communication Award from the Madri+d Knowledge Foundation for the article ‘ Is cancer hereditary ?’ ∞ Oriol Calvete has won the Juan Letona accésit prize awarded by the HM Hospitals Research Foundation for the best clinical translational medicine paper. Figure 2 Different strategies used in the polygenic analysis of families with testicular cancer. ANNUAL REPORT 2016 104 VICE-DIRECTION OF TRANSLATIONAL RESEARCH HEREDITARY ENDOCRINE CANCER GROUP Mercedes Robledo Group Leader Staff Scientists Alberto Cascón, Cristina Rodríguez SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 105 HUMAN CANCER GENETICS PROGRAMME | HEREDITARY ENDOCRINE CANCER GROUP OVERVIEW Our Group is mainly interested in identifying genetic risk factors involved in endocrine tumour susceptibility. Through a comprehensive analysis of tumour genomic features we have been able to propose diagnostic and prognostic markers, to identify altered pathways that could be therapeutically targeted, and to identify new major susceptibility genes. We are also interested in defining markers associated with differences in anticancer drug response and toxicity. We are applying targeted and whole-exome next-generation sequencing to a large series of clinically well-characterised patients. The aim is to identify new therapeutic approaches to personalise cancer treatment. These efforts will collectively improve the diagnosis, prognosis and treatment of patients. “ We identified a new major susceptibility gene for paraganglioma, a gene-net regulated by methylation in medullary thyroid carcinoma, and germline variants and tumour microRNAs that predict outcomes in cancer therapies.” Post-Doctoral Fellow Cristina Montero Graduate Students María V. Apellániz, Bruna Calsina, María Curras, Lucía Inglada, Veronika Mancikova ( until April ), Laura Remacha, Juan M. Roldán ( since February ) Technicians Rocío Letón, Rafael Torres ( TS )* *Titulado Superior ( Advanced Degree ) ANNUAL REPORT 2016 106 VICE-DIRECTION OF TRANSLATIONAL RESEARCH RESEARCH HIGHLIGHTS A gain-of-function mutation in DNMT3A causes paraganglioma The high percentage of patients carrying germline mutations makes pheochromocytomas ( PCC ) and paragangliomas ( PGL ) the most heritable of all tumours. However, there are still cases that are not explained by mutations in the known susceptibility genes. We aimed to identify the genetic cause in patients strongly suspected of having hereditary tumours. Whole-exome sequencing was applied to the germline of a parent- proband trio ( FIGURE ). Genome-wide methylome analysis of mutated tissues and targeted deep sequencing of 112 additional samples were also performed. Exome sequencing identified a single, novel de novo mutation in DNMT3A, DNA ( Cytosine- 5-)-Methyltransferase 3 Alpha, affecting a highly conserved residue located close to the aromatic cage responsible for binding the protein to trimethylated histone H3. DNMT3A-mutated tumour and blood tissue from the patient exhibited significant ( FDR<0.15 ) hypermethylation of homeobox-containing genes, providing evidence that the mutation plays an activating role. Targeted deep sequencing revealed the presence of subclonal mutations affecting the same residue in six additional PGLs, all of which exhibited positive staining for H3K9me3. The case described herein not only increases the number of known PCC/ PGL susceptibility genes, but also represents, to the best of our knowledge, the first example of a gain-of-function mutation affecting a DNA methyl transferase gene involved in cancer predisposition. Multilayer OMIC data in Medullary Thyroid carcinoma identifies the STAT3 pathway as a potential therapeutic target in RETM 918T tumours Medullary thyroid carcinoma ( MTC ) is a rare disease with few genetic drivers that, when diagnosed at an advanced stage, remains incurable. Due to its rarity, its genomic dissection has not been comprehensively explored. Exploiting multilayer genomic data, considering the transcriptome, miRNome and methylome, it was possible to uncover genes negatively regulated by methylation, such as DKK4, PLCB2, MMP20, miR-10a, miR- 30a and miR-200c, using MZ-CRC-1 and TT cell lines. Moreover, hypomethylation may induce activation of key pathways related to the malignant behaviour of RETM 918T-related MTCs. Functional annotation enrichment analysis identified the JAK/Stat pathway as a specific hallmark of RETM 918T-harbouring MTCs. In vitro studies with MTC cell models pointed to a RETM 918T genetic class-specific proliferative dependency on STAT3 activity. Remarkably, the inhibition of STAT3 increased the sensitivity of RETM 918T -bearing MTC cells to the FDA-approved RET inhibitor Vandetanib. This combinational treatment could potentially overcome the adverse effects encountered in clinical practice when Vandetanib monotherapy is applied. Identification of germline genetic variants and tumour microRNAs to predict outcomes in cancer therapies Personalised cancer treatment is of enormous clinical and social relevance since it can lead to safer and more efficient therapies. This year we focused our efforts on applying next generation sequencing to : i ) understand how low frequency genetic variants impact paclitaxel-induced neuropathy, and ii ) identify microRNAs predictive of the antiangiogenic drug response in renal cancer patients. Peripheral neuropathy diminishes the quality of life of many cancer patients, sometimes permanently, and limits the dose and efficacy of many cancer drugs. We found that low frequency variants in EPHA6, EPHA5 and EPHA8 genes contribute to the susceptibility to paclitaxel- induced neuropathy. Furthermore, EPHAs neuronal injury repair function suggests that these genes might constitute important neuropathy markers for many neurotoxic drugs. Regarding antiangiogenic therapies, these have drastically improved the survival of kidney cancer patients ; however, a fraction of the patients are refractory to these drugs. The first miRNome deep- sequencing study on an exceptional series of patients treated with sunitinib revealed microRNAs predictive of sunitinib response. Furthermore, a two microRNA-based classifier discriminated individuals with progressive disease upon sunitinib treatment ( P=1.3x10-4 ) with better predictive value than the commonly used clinicopathological risk factors. Thus, we provide new relevant markers that can help rationalise cancer treatment. s SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 107 HUMAN CANCER GENETICS PROGRAMME | HEREDITARY ENDOCRINE CANCER GROUP ∞ PUBLICATIONS ∞ Mancikova V, Montero-Conde C, Per- ales-Paton J, Fernandez AF, Santaca- na M, Jodkowska K, Inglada-Pérez L, Castelblanco E, Borrego S, Encinas M, Matias-Guiu X, Fraga MF, Robledo M ( 2016 ). Multilayer OMIC data in med- ullary thyroid carcinoma identifies the STAT3 pathway as a potential therapeutic target in RETM918T tumors. Clin Cancer Res. PMID : 27620278. ∞ Apellániz-Ruiz M, Tejero H, Inglada-Pérez L, Sánchez-Barroso L, Gutiérrez-Gutiérrez G, Calvo I, Castelo B, Redondo A, García- Donás J, Romero-Laorden N, Sereno M, Merino M, Currás-Freixes M, Monte- ro-Conde C, Mancikova V, Åvall-Lund- qvist E, Green H, Al-Shahrour F, Cascon A, Robledo M, Rodriguez-Antona C ( 2016 ). Targeted sequencing reveals low-frequen- cy variants in EPHA genes as markers of paclitaxel-induced peripheral neuropathy. Clin Cancer Res. PMID : 27582484. ∞ Smith J et al. ( incl. Robledo M ) ( 2016 ). Germline ESR2 mutation predisposes to medullary thyroid carcinoma and causes up-regulation of RET expression. Hum Mol Genet 25, 1836-1845. ∞ Benitez-Buelga C, Vaclová T, Ferreira S, Uri- oste M, Inglada-Perez L, Soberón N, Blasco MA, Osorio A, Benítez J ( 2016 ). Molecular insights into the OGG1 gene, a cancer risk modifier in BRCA1 and BRCA2 mutations carriers. Oncotarget 7, 25815-25825. ∞ Buj R et al. ( incl. Robledo M ) ( 2016 ). Quantification of Unmethylated Alu ( QUAlu ): a tool to assess global hypo- methylation in routine clinical samples. Oncotarget 7, 10536-10546. ∞ Richter S, Klink B, Nacke B, de Cubas AA, Mangelis A, Rapizzi E, Meinhardt M, Skondra C, Mannelli M, Robledo M, Menschikowski M, Eisenhofer G ( 2016 ). Epigenetic mutation of the Succinate De- hydrogenase C Promoter in a Patient with two Paragangliomas. J Clin Endocrinol Metab 101, 359-363. ∞ Diekstra MH, Belaustegui A, Swen JJ, Boven E, Castellano D, Gelderb- lom H, Mathijssen RH, García-Donas J, Rodríguez-Antona C, Rini BI, Guchelaar HJ ( 2016 ). Sunitinib induced hypertension in CYP3A4 rs4646437 A-allele carriers with metastatic renal cell carcinoma. Pharma- cogenomics J. PMID : 26810136. ∞ Duran I et al. ( incl. Rodríguez-Antona C ) ( 2016 ). SNPs associated with activity and toxicity of cabazitaxel in patients with advanced urothelial cell carcinoma. Pharmacogenomics 17, 463-471. ∞ Comino-Méndez I, Tejera ÁM, Cur- rás-Freixes M, Remacha L, Gonzalvo P, Tonda R, Letón R, Blasco MA, Robledo M, Cascón A ( 2016 ). ATRX driver mutation in a composite malignant pheochromocyto- ma. Cancer Genet 209, 272-277. ∞ Luque RM et al. ( incl. Robledo M ); investi- gadores del REMAH ( 2016 ). The Molecular Registry of Pituitary Adenomas ( REMAH ): A bet of Spanish Endocrinology for the future of individualized medicine and translational research. Endocrinol Nutr 63, 274-284. ∞ Guerrero Pérez F, Lisbona Gil A, Robledo M, Iglesias P, Villabona Artero C ( 2016 ). Pituitary adenoma associated with phe- ochromocytoma/paraganglioma : A new form of multiple endocrine neoplasia. Endocrinol Nutr 63, 506-508. ∞ Maison N, Korpershoek E, Eisenhofer G, Robledo M, de Krijger R, Beuschlein F ( 2016 ). Somatic RET mutation in a patient with pigmented adrenal pheochromocy- toma. Endocrinol Diabetes Metab Case Rep 2016, 150117. ∞ Maroñas O, Latorre A, Dopazo J, Pirmo- hamed M, Rodríguez-Antona C, Siest G, Carracedo Á, LLerena A ( 2016 ). Progress in pharmacogenetics : consortiums and new strategies. Drug Metab Pers Ther 31, 17-23. ∞ Rodriguez-Antona C ( 2016 ). The role of pharmacogenetics and pharmacogenom- ics in 21st-century medicine. Drug Metab Pers Ther 31, 1-2. ∞ Apellaniz-Ruiz M, Gallego C, Ruiz-Pinto S, Carracedo A, Rodríguez-Antona C ( 2016 ). Human genetics : international projects and personalized medicine. Drug Metab Pers Ther 31, 3-8. ∞ García-Donas J, Beuselinck B, Ingla- da-Pérez L, Graña O, Schöffski P, Wozniak A, Bechter O, Apellániz-Ruiz M, Lean- dro-García LJ, Esteban E, Castellano DE, González Del Alba A, Climent MA, Hernan- do S, Arranz JA, Morente M, Pisano DG, Robledo M, Rodriguez-Antona C ( 2016 ). Deep sequencing reveals microRNAs pre- dictive of antiangiogenic drug response. JCI Insight 7, e86051. ∞ AWARDS AND RECOGNITION ∞ Mercedes Robledo has received the Inter- national Medal awarded by the Society for Endocrinology. Figure Schematic representation of the coding variants passing all filtering steps ( with a genotype quality ≥ 90 ) that were found in the father ( orange square ), the mother ( green circle ), and the patient ( blue circle ) exomes, respectively. Manual curating by the integrative genomic viewer ( IGV ) excluded 4 of the 5 variants because they were either found in at least one of the progenitors or because they were probably the result of a sequencing artefact. The single de novo variant found ( c.896A>T in DNMT3 ) was validated by Sanger sequencing. ANNUAL REPORT 2016 108 VICE-DIRECTION OF TRANSLATIONAL RESEARCH GENETIC AND MOLECULAR EPIDEMIOLOGY GROUP Núria Malats Group Leader Staff Scientist M. Evangelina López De Maturana Post-Doctoral Fellows Paulina Gómez, M. Esther Molina Graduate Student Silvia Pineda ( until March ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 109 HUMAN CANCER GENETICS PROGRAMME | GENETIC AND MOLECULAR EPIDEMIOLOGY GROUP OVERVIEW The scope of research carried out by our Group ranges from the identification of aetiological agents and mechanisms, to the translation of the findings into the clinical and Public Health domains, focusing on bladder, pancreatic, and breast cancers. We employ a wide variety of biomarkers to better characterise exposures, genetic susceptibility patterns, and cancer outcomes. Omics data provide a unique opportunity in this regard and the Group explores its integration in epidemiologic studies. The strategic goals of the Group are to : ɗ Identify non-genetic and genetic factors, as well as their interactions, associated with cancer development and progression and with its molecular/omics subphenotypes. ɗ Develop and apply statistical/informatics tools to model the risk, prediction, and clinical course of patients with cancer by integrating epidemiologic with omics information. ɗ Assess clinical and public health strategies for cancer control using current genomic tests and data. “ We have undertaken in-depth analyses integrating omics and non- omics data to predict pancreatic and bladder cancer risk and outcome, and have assessed the challenges that epidemiology faces in this endeavour.” Technicians Ana Alfaro, Lola Alonso ( TS )*, Isabel Alonso ( since April ), Lidia Estudillo ( since April ), Mirari Márquez ( TS )*, Marta Rava ( TS )*, Rebeca Sánchez, Virginia Sebastián ( TS )* *Titulado Superior ( Advanced Degree ) Students in Practice Rianne Boenink ( Radboud University, Nijmegen, The Netherlands ), Isabel A. Martín ( National Institute of Health Carlos III, Madrid, Spain ), Ramón Tejedor ( National Institute of Health Carlos III, Madrid, Spain ), Mónica Cabrera ( Pompeu Fabra University, Barcelona, Spain ) Visiting Scientists M. Ángeles Jiménez ( until June ) ( Centro Nacional de Microbiología, National Institute of Health Carlos III, Spain ), Kristel Van Steen ( Liège University, Belgium ) ANNUAL REPORT 2016 110 VICE-DIRECTION OF TRANSLATIONAL RESEARCH RESEARCH HIGHLIGHTS Research findings During 2016, the Group mainly focussed its research on pancreatic and bladder cancers. Regarding pancreatic cancer ( PC ), we have further analysed the epidemiological and clinical data from the PanGenEU Study and have characterised the risk of PC associated with diabetes, multimorbidity patterns and family history of cancer, among others. We have completed the genome-wide association study ( GWAS ) and, in collaboration with the international consortia, we are now replicating the primary findings. We are exploring, in collaboration with experts in the field, genome-wide methylation data generated with the Illumina 850K array in cases and controls. We also participated in a study that identified 3 new pancreatic cancer susceptibility signals on chromosomes 1q32.1, 5p15.33 and 8q24.21. Regarding bladder cancer ( BC ), we showed that common SNPs have a limited role in predicting BC outcomes and reported, for the first time, a heritability estimate for disease outcome by assessing the predictive ability of the models, including up to 171,304 SNPs for tumour recurrence and progression. We have also provided proof of concept for the joint effect of genetic variants in improving the discriminative ability of clinical prognostic models by using innovative analytic approaches, and demonstrated that SNPs in inflammatory-related genes were associated with BC prognosis ( FIGURE 1 ). Through international collaborations, the Group has participated in the exploration of common germline variants in the APOBEC3 region associated with BC and breast cancer risk, and observed a tissue-specific role of environmental oncogenic triggers. In line with this study, mutations in cancer driver genes were primarily found in high-risk BC, together with APOBEC-related mutational signatures. We also participated in the development of a urine- based peptide biomarker and a combined methylation&mutation panel for detecting both primary and recurrent BC. Methodological contributions We have proposed an epidemiological-based integration of omics and non-omics data by considering the ‘ massive ’ inclusion of variables in the risk assessment and predictive models ( FIGURE 2 ). We also discussed the numerous challenges imbedding this type of research and have proposed analytical strategies that allow considering both omics and non-omics data used in the models towards a personalised prevention. Furthermore, we have adapted Bayesian sequential threshold models in combination with LASSO and applied them to time-to-event and the censoring nature of data, in order to study 822 BC patients followed-up more than 10 years. Translational activities We coordinate the COST Action BM1204 EUPancreas ( www. eupancreas.com ). This Action includes 250 multidisciplinary members from 22 EU countries, EU governmental and nongovernmental institutions, and private companies. Several scientific, training, and dissemination activities have been conducted during 2016. By endorsing the Pancreatic Cancer Europe ( PCE ) multi-stakeholder platform, we have actively participated in several activities aimed at increasing the awareness of PC in the general population, the medical community, and among health policy makers. The Group has actively participated in setting up a European-based clinical registry of PC ( PancreOS ) jointly with the EPC, the Joint Research Centre from the European Community, and the European Network of Cancer Registries. The Group has also contributed to the development of recommendations for a state strategy for personalised/precision medicine, led by the Roche Institute. Another area our Group contributed to was the identification of different sources of big data and the importance of unstructured data for potential future uses in drug discovery ; the main practical and ethical challenges to unravel the full potential of big data in healthcare were discussed. s Figure 1 Progression-free survival of the 822 non-muscle invasive BC patients according to CD3G-rs3212262 genotypes. Five-year progression free survival was 92% for AA, 85% for Aa and 71% for aa genotypes ( log rank p-value=8.4x10-4, adjusted Cox p-value = 0.023 ). SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 111 HUMAN CANCER GENETICS PROGRAMME | GENETIC AND MOLECULAR EPIDEMIOLOGY GROUP ∞ PUBLICATIONS ∞ Middlebrooks CD et al. ( incl. Malats N ) ( 2016 ). Germline variants in the APOBEC3 region predispose to cancer and APOBEC mutation signature in tumors. Nat Genet 48, 1330-1338. ∞ Zhou W et al. ( incl. Real FX, Malats N ) ( 2016 ). Mosaic loss of chromosome Y is associated with common variation near TCL1A. Nat Genet 48, 563-568. ∞ Hedegaard J et al. ( incl. Malats N, Real FX ) ( 2016 ). Comprehensive Transcrip- tional Analysis of Early Stage Urothelial Carcinoma. Cancer Cell 30, 27-42. ∞ Martinelli P, Carrillo-de Santa Pau E, Cox T, Sainz B, Dusetti N, Greenhalf W, Rinaldi L, Costello E, Ghaneh P, Malats N, Buchler M, Iovanna J, Neoptolemos J, Real FX ( 2016 ). GATA6 regulates EMT, metastatic potential and response to chemotherapy in pancreatic cancer. Gut. PMID : 27325420. ∞ Machiela MJ et al. ( incl. Real FX, Malats N ) ( 2016 ). Female chromosome x mo- saicism is age-related and preferentially affects the inactivated x chromosome. Nat Commun 7, 11843. ∞ Frantzi M et al. ( incl. Malats N ) ( 2016 ). De- velopment and validation of urine-based peptide biomarker panels for detecting bladder cancer in a multi-center study. Clin Cancer Res 22, 4077-4086. ∞ Li Z, Rava M, Bédard A, Dumas O, Gar- cia-Aymerich J, Leynaert B, Pison C, Le Moual N, Romieu I, Siroux V, Camargo CA Jr, Nadif R, Varraso R ( 2016 ). Cured meat intake is associated with worsening asth- ma symptoms. Thorax. PMID : 27999171. ∞ Shi J et al. ( incl. Malats N ) ( 2016 ). Winner’s curse correction and variable threshold- ing improve performance of polygenic risk modeling based on genome-wide association study summary-level data. PLoS Genet. PMID : 28036406. ∞ Figueroa JD et al. ( incl. Malats N ) ( 2016 ). Identification of a novel susceptibility lo- cus at 13q34 and refinement of the 20p12.2 region as a multi-signal locus associated with bladder cancer risk in Europeans. Hum Mol Genet 25, 1203-1214. ∞ Zhang M et al. ( incl. Malats N ) ( 2016 ). Three new pancreatic cancer susceptibility signals identified on chromosomes 1q32.1, 5p15.33 and 8q24.21. Oncotarget. PMID : 27579533. ∞ Beukers W et al. ( incl. Real FX, Malats N ) ( 2016 ). FGFR3, TERT and OTX1 as urinary biomarker combination for surveillance of bladder cancer patients in a large pro- spective multicenter study. J Urol. PMID : 28049011. ∞ van Kessel KE et al. ( incl. Real FX, Malats N ) ( 2016 ). Validation of a DNA meth- ylation-mutation urine assay to select patients with hematuria for cystoscopy. J Urology. PMID : 27746284. ∞ Masson-Lecomte A *, López de Matura- na E *, Goddard ME, Picornell A, Rava M, González-Neira A, Márquez M, Carrato A, Tardon A, Lloreta J, Garcia-Closas M, Silverman D, Rothman N, Kogevinas M, Allory Y, Chanock S, Real FX, Malats N, on behalf of the SBC/EPICURO Study In- vestigators ( 2016 ). Inflammatory-related genetic variants in non-muscle invasive bladder cancer prognosis. A multi-marker Bayesian assessment. Cancer Epidemiol Biomarkers Prev 25,1144-1150. ∞ Carrión-García CJ, Guerra-Hernández EJ, García-Villanova B, Molina-Montes E ( 2016 ). Non-enzymatic antioxidant capacity ( NEAC ) estimated by two dif- ferent dietary assessment methods and its relationship with NEAC plasma levels. Eur J Nutr. PMID : 27023743. ∞ López de Maturana E et al. ( incl.Real FX, Malats N ); On behalf of the SBC/EPICURO Study Investigators ( 2016 ). Prediction of non-muscle invasive bladder cancer out- comes assessed by innovative multimarker prognostic models. BMC Cancer 16,351. ∞ Matulonga B, Rava M, Siroux V, Bernard A, Dumas O, Pin I, Zock JP, Nadif R, Ley- naert B, Le Moual N ( 2016 ). Women using bleach for home cleaning are at increased risk of non-allergic asthma. Resp Med 117, 264-271. ∞ Witjes JA, et al. ( incl. Malats N ) ( 2016 ). The efficacy of BCG Tice and BCG Con- naught in a cohort of 2099 T1G3 non-mus- cle invasive bladder cancer patients. Urol Oncol 34,484. ∞ Leyens L, Reumann M, Malats N, Brand A ( 2016 ). Use of Big Data for drug de- velopment and for Public and Personal Health and Care. Genet Epidemiol. PMID : 27873357. ∞ Lopez de Maturana E, Pineda S, Brand A, Malats N ( 2016 ). Towards the integration of Omics data in epidemiological studies : still a “ long and winding road ”. Genet Epidemiol 40, 558-569. ∞ Friesen MC et al. ( incl. Malats N ) ( 2016 ). Combining Decision Rules from Classifica- tion Tree Models and Expert Assessment to Estimate Occupational Exposure to Diesel Exhaust for a Case-Control Study. Ann Occup Hyg 60, 467-478. ∞ PATENT ∞ Allory Y, Kamoun A, Vlahou A, Makrydakis M, Zoidakis I ; Masselon C, Garin J, Court M, Malats N, Marquez Cid M, Domon B, Duriez E ; Radvanyi F. ( 2016 ). Bladder Cancer Biomarker Proteins. PCT/EP 15 202 453.5. ∞ AWARDS AND RECOGNITION ∞ Founder and Board Member of Pancreatic Cancer Europe. ∞ Member of the Working Group ‘ Recomen- daciones para un plan de Medicina de Pre- cisión ’, Fundación Instituto Roche, Spain. Figure 2 Conceptual association models applied in classical (  A ), molecular ( B ), genetic ( C ) and omics integrative epidemiology ( D, E, F, and G ). ANNUAL REPORT 2016 112 VICE-DIRECTION OF TRANSLATIONAL RESEARCH FAMILIAL CANCER CLINICAL UNIT OVERVIEW Individuals that present with an uncommon malignancy or with cancer at an early age of onset deserve special attention because they are more likely to harbour an inherited predisposition and may require unique treatment strategies. Identification of a heritable cancer predisposition syndrome is not only essential for genetic counselling and for the design of a surveillance scheme for both the patient and his/her relatives, but also for facilitating the refinement and optimisation of treatment strategies so as to minimise toxicity and maximise efficacy. Vigilance of these syndromes can significantly enhance the quality and comprehensiveness of clinical management of cancer. In addition, the evaluation of inherited cancer predisposition is changing with the routine use of NGS. Despite the promise of NGS, the utility of testing multiple genes with different modes of inheritance and with varying levels of penetrance has been questioned due to the increasing costs of surveillance and unnecessary treatments, and the uncertain consequences of the identification of variants of unknown significance. More than ever it is necessary to underline that NGS testing should only be offered in the context of expert genetic counselling. In the Cancer Genetics Consultation of the Familial Cancer Clinical Unit ( FCCU ) we work together with Fuenlabrada Hospital clinicians, as well as health-care providers from other Madrid hospitals and other Autonomous Communities, in order to heighten the vigilance of hereditary cancer syndromes and to better adapt the genetic counselling process in alignment with the introduction of new technologies. Miguel Urioste Clinical Unit Head Graduate Student Laura Pena Technicians Maika González, Fátima Mercadillo SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 113 HUMAN CANCER GENETICS PROGRAMME | FAMILIAL CANCER CLINICAL UNIT CLINICAL, DIAGNOSTIC AND RESEARCH HIGHLIGHTS The FCCU evaluates individuals and families who are at an increased risk of developing cancer at our cancer genetics consultancy in the Medical Oncology Service of the Hospital Universitario de Fuenlabrada ( HUF ). The referral system, surveillance and treatment measures are discussed with medical oncologists and other clinicians during the monthly sessions conducted by the hospital’s Hereditary Cancer Clinical Committee. During 2016, our consultancy at HUF was visited by 408 patients, a 21% increase over 2015. Moreover, 352 genetic diagnostic studies were performed in the FCCU laboratory ( 306 in 2015 ). We also tested patients with multiple gene panels ; this enables us to offer results on genes of interest in just a few weeks ’ time. The FCCU has continued to actively contribute to unravel the complexity of genetic cancer predisposition and to help refine tools for a better evaluation of patients and families. FCCU members collaborate with the ‘ Lynch Syndrome prediction model validation study group ’ to define the most precise tools for the evaluation of families with hereditary colorectal cancer as well as to identify the best candidates for genetic studies. In collaboration with other research groups, the FCCU has defined the role of the UNC5C gene in hereditary forms of colorectal cancer and in polyposis, as well as the role of OGG1 as a cancer risk modifier in BRCA1 and 2 mutation carriers. Genetic susceptibility to colorectal cancer is a key area focus for the FCCU’s research activities. Familial or hereditary forms of colorectal cancer, early-onset colorectal cancer ( EOCC ), and synchronous or metachronous colorectal tumours are our main topics of interest. We have continued the characterisation of EOCC, on the premise that the carcinogenetic mechanism and the progression of these tumours may differ in comparison with late-onset colorectal cancer ( LOCC ) ( FIGURE ). The APC gene status, wild-type or mutated, seems to be a marker of prognosis in colorectal cancer with microsatellite stability ( MSS ), but the prognosis would have a different sign in EOCC and LOCC. In MSS- EOCC, the worst prognosis was associated with APC-mutated tumours and distal location. However, in the MSS-LOCC group, the worst prognosis was observed among proximally located tumours with APC-wild type. These results not only continue to suggest a different behaviour according to the age of onset, but also define different groups in relation to the tumour location. During 2016 the FCCU has maintained a fruitful relationship with AEAS. Several members of the association have received genetic counselling in our consultancy, and the study of sarcoma predisposition genes ( mainly TP53, POT1 and CDKN2A ) was also carried out in our laboratory. These activities are part of our ongoing collaborations with cancer patients associations. Recently, we have designed a new survey that will be distributed among members of the AEFAT with the aim of identifying those families with an increased susceptibility to cancer. s ∞ PUBLICATIONS ∞ Kastrinos F et al. ( incl. Urioste M ); Lynch Syndrome prediction model validation study group ( 2016 ). Comparison of Prediction Models for Lynch Syndrome Among Individuals With Colorectal Can- cer. J Natl Cancer I 108, djv308. ∞ Vaclová T et al. ( incl. Megías D, Gomez- Lopez S, Barroso A, Urioste M, Benítez J, Osorio A ) ( 2016 ). Germline missense pathogenic variants in the BRCA1 BRCT domain, p.Gly1706Glu and p.Ala1708G- lu, increase cellular sensitivity to PARP inhibitor Olaparib by a dominant neg- ative effect. Hum Mol Genet. PMID : 27015555. ∞ Benitez-Buelga C, Vaclová T, Ferreira S, Uri- oste M, Inglada-Perez L, Soberón N, Blasco MA, Osorio A, Benitez J ( 2016 ). Molecular insights into the OGG1 gene, a cancer risk modifier in BRCA1 and BRCA2 mutations carriers. Oncotarget 7, 25815-25825. ∞ Mur P et al. ( incl. Urioste M ) ( 2016 ). Scarce evidence of the causal role of germline mutations in UNC5C in hereditary colorec- tal cancer and polyposis. Sci Rep-UK 6, 20697. ∞ Perea J et al. ( incl. Urioste M ) ( 2016 ). Comment on ‘ Wild-type APC predic- tion of poor prognosis in microsatel- lite-stable proximal colorectal cancer differs according to the age of onset ’. Br J Cancer 114, e7. ∞ Arriba M et al. ( incl. Urioste M ) ( 2016 ). Unsupervised analysis of array compar- ative genomic hybridization data from early-onset colorectal cancer reveals equivalence with molecular classification and phenotypes. Neoplasia, 19,28-34. ∞ Arriba M et al. ( incl. Urioste M ) ( 2016 ). Toward a Molecular Classification of Syn- chronous Colorectal Cancer : Clinical and Molecular Characterization. Clin Cancer Colorectal. PMID : 27600984. ∞ Perea J et al. ( incl. Urioste M ) ( 2016 ).Fre- quency and impact of KRAS mutations in early-onset colorectal cancer. Hum Pathol. PMID : 27816716. Book chapter ∞ Rueda Fernández D, Urioste Azcorra M ( 2016 ). Biología Molecular en el cáncer he- reditario. In : Manual de Genética y Biología Molecular. Ed : AEBM - Medicina de Labo- ratorio. Madrid. ISBN : 978-84-617-4821-1. ∞ PATENT ∞ Perea García J, González-Sarmiento R, Urioste Azcorra M, Rueda Fernández D, Arriba Doménech M, García Hernández JL, Pérez García J ( 2016 ). Biomarcador para el diagnóstico, pronóstico y seguimiento de cáncer colorrectal de aparición precoz. PCT/ES2016/070929. Figure Copy number gains and losses in ≤45 y-o and ≥70 y-o colorectal cancers. ANNUAL REPORT 2016 114 VICE-DIRECTION OF TRANSLATIONAL RESEARCH MOLECULAR CYTOGENETICS AND GENOME EDITING UNIT OVERVIEW Chromosomal translocations are very common events involved in the development of several cancers, especially in sarcomas and haematological malignancies. The research activity of the Molecular Cytogenetics and Genome Editing Unit covers the main topics related to cancer cytogenetics and genome engineering : from classical cytogenetics techniques to new genome engineering tools, including the CRISPR-Cas9 system. We are focusing on the implementation and development of new technologies to enhance knowledge about the biology of tumours and to discover new potential therapeutic targets. With the combined use of CRISPR-Cas9 genome editing and cellular technologies, we are creating in vitro models that recapitulate chromosomal and genetic cancer alterations. Members of the Unit also participate in collaborative projects with clinical and basic science investigators across the CNIO and other institutes. “ By way of different molecular approaches, we generate human cancer cell models carrying tumour-associated chromosomal translocations in order to study their functional contribution to oncogenesis.” Juan C. Cigudosa Unit Head Staff Scientist Sandra Rodríguez SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 115 HUMAN CANCER GENETICS PROGRAMME | MOLECULAR CYTOGENETICS AND GENOME EDITING UNIT RESEARCH HIGHLIGHTS Optimising CRISPR-Cas9 to model cancer aberrations in primary cells In vitro modelling of complex tumour-associated chromosome translocations at native loci is feasible with CRISPR. However, the generation of translocations must be optimised, especially for mimicking events in human primary cells. We have optimised our CRISPR protocol to efficiently obtain those cells, thereby enabling the rescue of translocation+ populations of human primary cells, including induced pluripotent stem ( iPS ) cells and mesenchymal stem cells ( MSCs ). These models can surely help us to understand the molecular mechanisms underlying the initiation of human cancers, and can also be used for high-throughput drug screening, toxicological testing and biomarker identification. From the patient’s chromosome translocations to their functional effects We have worked on the oncogenic role of the translocation t( 8 ;21 )( q22 ;q22 )/RUNX1-RUNX1T1, which occurs in 4% of acute myeloid leukaemia patients. We deciphered a new function for the activation of MAPK8, observed in t( 8 ;21 )+ cells, which is responsible for the stabilisation of SP1. Our data show the essential role of SP1 in t( 8 ;21 )+ cell maintenance through the regulation of key genes, such as CDKN1A. These results provide new evidence for the inclusion of pharmacological approaches leading to degradation of SP1 in the treatment of these patients. Technological and translational activities We provide state-of-the-art molecular cytogenetics and genome editing services. The Unit makes available various techniques to the CNIO Research Groups ; these techniques provide more sensitive and accurate tools to analyse cancer cells, such as RNA-FISH, chromosome stability studies based on a combined array CGH-FISH approach, or the use of CRISPR libraries to perform high-throughput functional analysis. For gene editing experiments, we have set up a specific FISH analysis to detect genomic integration sites of small constructs including LV particles. In 2016, we carried out over 1,000 assays for experimental and clinically-oriented projects. s Post-Doctoral Fellow Raúl Torres Technicians Angelo Bertini ( until April ) ( TS )*, M. Carmen Carralero, M. Carmen Martín, Marta Martínez-Lage ( since June ) ( TS )*, Francisco J. Moya ( TS )* *Titulado Superior ( Advanced Degree ) ∞ PUBLICATIONS ∞ Richart L, Carrillo-de Santa Pau E, Río- Machín A, de Andrés MP, Cigudosa JC, Lobo VJ, Real FX. BPTF is required for c-MYC transcriptional activity and in vivo tumorigenesis ( 2016 ). Nat Commun 7, 10153. ∞ Rodriguez-Perales S, Torres-Ruiz R, Suela J, Acquadro F, Martin MC, Yebra E, Ramirez JC, Alvarez S, Cigudosa JC ( 2016 ). Trun- cated RUNX1 protein generated by a novel t( 1 ;21 )( p32 ;q22 ) chromosomal translocation impairs the proliferation and differentiation of human hematopoi- etic progenitors. Oncogene 35, 125-134. ∞ Muñoz-López A, Romero-Moya D, Prieto C, Ramos-Mejía V, Agraz-Doblas A, Varela I, Buschbeck M, Palau A, Carvajal-Vergara X, Giorgetti A, Ford A, Lako M, Granada I, Ruiz-Xivillé N, Rodríguez-Perales S, Tor- res-Ruíz R, Stam RW, Fuster JL, Fraga MF, Nakanishi M, Cazzaniga G, Bardini M, Cobo I, Bayon GF, Fernandez AF, Bueno C, Menendez P ( 2016 ). Development refrac- toriness of MLL-rearranged human B cell acute leukemias to reprogramming into pluripotency. Stem Cell Reports 7, 602-618. ∞ Maiques-Diaz A, Hernando M, Sánchez- López A, Rio-Machin A, Shrestha M, Mulloy JC, Cigudosa JC, Alvarez S ( 2016 ). MAPK8-mediated stabilization of SP1 is essential for RUNX1-RUNX1T1 - driven leukaemia. Br J Haematol 172, 807-810. ∞ Torres-Ruiz R, Rodriguez-Perales S ( 2016 ). CRISPR-Cas9 technology : applications and human disease modelling. Brief Funct Genomics. PMID : 27345434 Figure Targeted chromosomal translocations workflow using CRISPR ribonucleoprotein (  RNP ), GFP molecule controls and single-stranded oligodeoxynucleotides ( ssODNs ) transfection approach. ANNUAL REPORT 2016 116 VICE-DIRECTION OF TRANSLATIONAL RESEARCH HUMAN GENOTYPING- CEGEN UNIT OVERVIEW The most abundant types of genetic variation are single nucleotide variants ( SNVs ) and copy number variants ( CNVs ). Association studies involving the large-scale analysis of both SNVs and CNVs in thousands of patients can help to identify genes underlying complex diseases such as cancer, and drug responses. In this Unit we implement different high-throughput and cost-effective methods to measure from one to millions of SNVs and CNVs. In addition, epigenetic studies using whole-genome methylation arrays are performed in the Unit. Complementarily, research focused on the identification of biomarkers for precision medicine is also undertaken. RESEARCH HIGHLIGHTS Identification of genetic variants associated with docetaxel and anthracycline efficacy Taxanes and anthracyclines are widely used in the treatment of breast cancer, despite the benefit being limited to a small proportion of patients and that preoperative biomarkers, which are predictive of clinical outcome, still remain lacking. We carried out a pharmacogenetic study in 181 patients with locally advanced breast cancer, previously enrolled in a phase 2 randomised clinical trial ( NCT00123929 ), in which patients were randomly assigned to receive doxorubicin ( anthracycline ) or docetaxel ( taxane ) neoadjuvant chemotherapy. We assessed whether genetic variants in 15 key transport or metabolism genes relevant to doxorubicin and docetaxel drugs could play a role as predictive biomarkers. We identified a genetic variant located in Anna González Neira Unit Head Graduate Student Sara Ruiz “ Advances in understanding patients ’ responses to therapy will help to individualise cancer patient care.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 117 HUMAN CANCER GENETICS PROGRAMME | HUMAN GENOTYPING-CEGEN UNIT the promoter of ABCC2 as the strongest association with tumour response in patients treated with doxorubicin ( P=0.009 ). We also identified a significant association for an intronic variant located in CYP1B1 associated with docetaxel tumour response ( P=2.15x10-⁴ ). Our integrated pathway-based approach enables the revealing of promising genetic biomarkers of treatment outcome in breast cancer patients. New low-frequency variant loci associated with anthracycline-induced cardiotoxicity ( AIC ) in cancer patients by Illumina HumanExome Beadchip Anthracycline chemotherapeutic agents are widely used in the treatment of cancer ; however, chronic anthracycline-induced cardiotoxicity ( AIC ) is a serious long-term complication leading to substantial morbidity. Our aim was to identify new genes and low-frequency variants influencing the susceptibility to AIC. We studied the association of variants on the Illumina HumanExome BeadChip array in a discovery cohort of breast cancer anthracycline-treated patients. Using gene-based tests ( SKAT-O ) that have greater statistical power to detect rare variant associations and that can evaluate the cumulative effect of multiple genetic variants, we identified novel significant associations in a gene with a major role in mitochondrial fatty acid β-oxidation and the respiratory chain, involved in anthracycline- related toxicity via an oxidative stress mechanism. We replicated our association results in another cohort of anthracycline treated paediatric cancer patients from Spain. Functional characterisation at the 20q13.33 risk locus for capecitabine-induced hand-foot syndrome ( CiHFS ) Capecitabine is a chemotherapy drug widely used in breast and colorectal cancer ; the most frequent adverse drug reaction to this treatment ( in 30% of the patients ) is CiHFS, a cause of dose reductions and dose delays. By genome-wide association studies ( GWAS ), we identified four linked CDH4 regulatory variants ( h2=risk haplotype ) associated with the risk of CiHFS appearance ( HR=2.48 p=1.43x10- 8 ). The CDH4 gene encodes R-Cadherin, which is localised in the granular layer of the epidermis and is involved in the cohesiveness of epithelial layers. We demonstrated that these regulatory variants are able to mediate chromatin structural changes in chromatin organisation, which results in the presence of the risk alleles and in decreased expression levels of CDH4 mRNA and R-Cadherin protein. Additional functional experiments are being performed. The study has been carried out in collaboration with CNIO’s Chromosome Dynamics Group and the Epithelial Cell Biology Group. s Technicians Charo Alonso, Núria Álvarez, Belén Herráez, Tais Moreno ( until October ) ( TS )*, Guillermo Pita ( TS )*, Rocío Nuñez ( since October ) ( TS )* *Titulado Superior ( Advanced Degree ) ∞ PUBLICATIONS ∞ Dunning AM et al. ( incl. González-Neira A, Osorio A, Benítez J ) ( 2016 ). Breast cancer risk variants at 6q25 display different pheno- type associations and regulate ESR1, RMND1 and CCDC170. Nat Genet 48, 374-386. ∞ Kar SP et al. ( incl. González Neira A, Benítez J ) ( 2016 ). Genome-wide me- ta-analyses of breast, ovarian, and pros- tate cancer association studies identify multiple new susceptibility loci shared by at least two cancer yypes. Cancer Discov- ery 6, 1052-1057. ∞ Couch FJ et al. ( incl. González-Neira A, Osorio A, Benítez J ) ( 2016 ). Identification of four novel susceptibility loci for oes- trogen receptor negative breast cancer. Nat Commun 7, 11375. ∞ Lawrenson K et al. ( incl. González - Neira A ) ( 2016 ). Functional mechanisms under- lying pleiotropic risk alleles at the 19p13.1 breast-ovarian cancer susceptibility locus. Nat Commun 7, 12675. ∞ Ghoussaini M et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Evidence that the 5p12 Variant rs10941679 Confers Susceptibili- ty to Estrogen-Receptor-Positive Breast Cancer through FGF10 and MRPS30 Regulation. Am J Hum Genet 99, 903-911 ∞ Zagorac S et al. ( incl. González - Neira A ) ( 2016 ). DNMT1 Inhibition Reprograms Pancreatic Cancer Stem Cells via Upreg- ulation of the miR-17-92 Cluster. Cancer Res 76, 4546-4558. ∞ Ruiz-Pinto S et al. ( incl. González - Neira A ) ( 2016 ). Identification of genetic vari- ants in pharmacokinetic genes associated with Ewing Sarcoma treatment outcome. Ann Oncol 27, 1788-1793. ∞ Wyszynski A et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). An intergenic risk locus containing an enhancer deletion in 2q35 modulates breast cancer risk by deregulating IGFBP5 expression. Hum Mol Genet 25, 3863-3876. ∞ Hamdi Y et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Association of breast cancer risk with genetic variants showing differential allelic expression : Identifica- tion of a novel breast cancer susceptibility locus at 4q21. Oncotarget 7, 80140-80163. ∞ Southey MC et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). PALB2, CHEK2 and ATM rare variants and cancer risk : data from COGS. J Med Genet 53, 800-811. ∞ Easton DF et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). No evidence that protein truncating variants in BRIP1 are associated with breast cancer risk : im- plications for gene panel testing. J Med Genet 53, 298-308. ∞ Petridis C et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Genetic predisposition to ductal carcinoma in situ of the breast. Breast Cancer Res 18, 22. ∞ Zeng C et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Identification of inde- pendent association signals and putative functional variants for breast cancer risk through fine-scale mapping of the 12p11 locus. Breast Cancer Res 18, 64. ∞ Darabi H et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Fine scale mapping of the 17q22 breast cancer locus using dense SNPs, genotyped within the Collaborative Oncological Gene-Environment Study ( COGs ). Sci Rep 7, 32512. ∞ Liu J et al. ( incl. Benítez J, González-Neira A ) ( 2016 ). rs2735383, located at a microR- NA binding site in the 3 ’UTR of NBS1, is not associated with breast cancer risk. Sci Rep 6, 36874. ∞ Lei J et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Genetic variation in the immu- nosuppression pathway genes and breast cancer susceptibility : a pooled analysis of 42,510 cases and 40,577 controls from the Breast Cancer Association Consortium. Hum Genet 135, 137-154. ∞ Masson-Lecomte A et al . (   incl . González-Neira A, Real FX, Malats N ) ( 2016 ). Inflammatory-Related Genetic Variants in Non-Muscle-Invasive Bladder Cancer Prognosis : A Multimarker Bayesian Assessment. Cancer Epidemiol Biomark 25, 1144-1150. ∞ Horne HN et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). Fine-Mapping of the 1p11.2 Breast Cancer Susceptibility Locus. PLoS One 11, e0160316. ∞ Pelttari LM et al. ( incl. González-Neira A, Benítez J ) ( 2016 ). RAD51B in Familial Breast Cancer. PLoS One 11, e0153788. ANNUAL REPORT 2016 118 VICE-DIRECTION OF TRANSLATIONAL RESEARCH CLINICAL RESEARCH PROGRAMME MANUEL HIDALGO ( Until December ) Programme Director SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 119 CLINICAL RESEARCH PROGRAMME The Clinical Research Programme ( CRP ) aims to translate advances in cancer research into the prevention, diagnosis, and treatment of patients. The major goals of the CRP are the conduction of early clinical trials with novel drugs, the discovery of biomarkers of drug action and disease outcome, the implementation of a strategy for personalised medicine, and the launching of a training programme in drug development. The CRP is composed of 5 Clinical Research Units ( CRU ) and 1 support Unit. The Gastrointestinal Cancer CRU, led by Manuel Hidalgo until December 2016, studies novel therapeutics and personalised medicine in pancreatic cancer. Miguel Quintela- Fandino leads the Breast Cancer CRU that works on the development of kinase and angiogenesis inhibitors in breast cancer, as well as on the understanding of the molecular taxonomy and metabolic vulnerabilities of this disease. The Prostate Cancer CRU, led by David Olmos, explores novel therapeutics and biomarkers of the disease, with a particular interest in understanding DNA damage repair deficiency mechanisms in prostate cancer. The Lung Cancer CRU, headed by Luis Paz- Ares, and the Haematological Malignancies CRU, led by Joaquín Martínez-López – both established as part of an agreement with the Hospital Universitario 12 de Octubre − focus on molecular and preclinical studies in non-small cell lung cancer and in multiple myeloma, respectively. The Molecular Diagnostics Unit, led by Luis Lombardía, provides support to medical professionals of the National Health system and the CRP through the provision of a wide variety of molecular tests that determine alterations in biomarkers involved in cancer. In 2016, the Programme continued the expansion of its clinical trials activities in collaboration with several hospitals in Spain. “ The Clinical Research Programme focuses on developing novel and more effective treatments against cancer.” ANNUAL REPORT 2016 120 VICE-DIRECTION OF TRANSLATIONAL RESEARCH GASTROINTESTINAL CANCER CLINICAL RESEARCH UNIT Manuel Hidalgo ( until December ) Clinical Research Unit Head Staff Scientists Rodrigo De Almeida, Pedro P. López, Sofía Perea, Begoña Vázquez ( since April ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 121 CLINICAL RESEARCH PROGRAMME | GASTROINTESTINAL CANCER CLINICAL RESEARCH UNIT OVERVIEW The Gastrointestinal ( GI ) Cancer Clinical Research Unit focuses on the clinical development of novel therapeutics for patients with cancers of the gastrointestinal tract as well as personalised medicine approaches for these patients. The work of the Group combines the preclinical assessment of novel anticancer agents in ‘ Avatar ’ mouse models with the design, conduction, and analysis of clinical trials with novel anticancer agents in patients with gastrointestinal tumours. Over the last few years the Group has implemented a growing portfolio of clinical trials with new agents spanning a broad range of mechanisms of action. Key to the work is the development and characterisation of Avatar mouse models for drug screening, biomarker development, and personalised medicine. The Group has developed and has characterised the largest collection of these models in pancreatic cancer. Avatar models are used in 3 critical applications : ( i ) the screening of new anticancer agents ; ( ii ) conduction of co-clinical trials, in which ongoing clinical trials are performed in parallel with studies using Avatar models of the same cancer type in order to elucidate mechanisms of action and biomarkers of drug response/resistance ; and ( iii ) finally, the Avatar models for personalised cancer treatment integrated with next generation sequencing. s Clinical Investigators M José De Miguel ( until June ), Laura Medina ( TS )* Post-Doctoral Fellows Lucía Fernández ( until June ), Camino Menéndez, María Vela ( until February ) Graduate Students Spas Dimitrov, Beatriz Salvador Technicians Carolina Alonso ( since December ), Soraya Ardila ( until March ) ( TS )*, Natalia Baños, Victoria B.bonilla, Yolanda Durán, Manuel Muñoz, Gemma M. Sánchez ( TS )*, Francesca Sarno ( TS )* *Titulado Superior ( Advanced Degree ) Visiting Scientists Raul Calero ( CNIO-HNJ Clinical Research Unit, Hospital Infantil Univesitario Niño Jesús, Madrid ), Lucas Moreno ( CNIO-HNJ Clinical Research Unit, Hospital Infantil Univesitario Niño Jesús, Madrid ) ∞ PUBLICATIONS ∞ Ambrogio C, Gómez-López G, Falcone M, Vidal A, Nadal E, Crosetto N, Blasco RB, Fernández-Marcos PJ, Sánchez-Céspedes M, Ren X, Wang Z, Ding K, Hidalgo M, Serrano M, Villanueva A, Santamaría D, Barbacid M ( 2016 ). Combined inhibition of DDR1 and Notch signaling is a ther- apeutic strategy for KRAS-driven lung adenocarcinoma. Nat Med 22, 270-277. ∞ Toledo RA, Cubillo A, Vega E, Garralda E, Alvarez R, de la Varga LU, Pascual JR, Sánchez G, Sarno F, Prieto SH, Perea S, Lopéz-Casas PP, López-Ríos F, Hidalgo M ( 2016 ). Clinical validation of prospec- tive liquid biopsy monitoring in patients with wild-type RAS metastatic colorectal cancer treated with FOLFIRI-cetuximab. Oncotarget. PMID : 27852040. ∞ Hernández-Agudo E, Mondejar T, So- to-Montenegro ML, Megías D, Mouron S, Sanchez J, Hidalgo M, Lopez-Casas PP, Mulero F, Desco M, Quintela-Fandino M. ( 2016 ). Monitoring vascular normalization induced by antiangiogenic treatment with ( 18 )F-fluoromisonidazole-PET. Mol Oncol 10, 1704-1718. ∞ Céspedes MV, Guillén MJ, López-Casas PP, Sarno F, Gallardo A, Álamo P, Cuevas C, Hidalgo M, Galmarini CM, Allavena P, Avilés P, Mangues R ( 2016 ). Lurbinectedin induces depletion of tumor-associated macrophages, an essential component of its in vivo synergism with gemcitabine, in pancreatic adenocarcinoma mouse mod- els. Dis Model Mech 9, 1461-1471. ∞ Rajeshkumar NV, Yabuuchi S, Pai SG, Tong Z, Hou S, Bateman S, Pierce DW, Heise C, Von Hoff DD, Maitra A, Hidalgo M ( 2016 ). Superior therapeutic efficacy of nab-pa- clitaxel over cremophor-based paclitaxel in locally advanced and metastatic models of human pancreatic cancer. Br J Cancer 115, 442-453. ∞ Hidalgo M, Álvarez R, Gallego J, Guillén- Ponce C, Laquente B, Macarulla T, Muñoz A, Salgado M, Vera R, Adeva J, Alés I, Arévalo S, Blázquez J, Calsina A, Carmona A, de Madaria E, Díaz R, Díez L, Fernández T, de Paredes BG, Gallardo ME, González I, Hernando O, Jiménez P, López A, López C, López-Ríos F, Martín E, Martínez J, Martínez A, Montans J, Pazo R, Plaza JC, Peiró I, Reina JJ, Sanjuanbenito A, Yaya R, Carrato A ( 2016 ). Consensus guidelines for diagnosis, treatment and follow-up of patients with pancreatic cancer in Spain. Clin Transl Oncol. PMID : 27995549. ANNUAL REPORT 2016 122 VICE-DIRECTION OF TRANSLATIONAL RESEARCH BREAST CANCER JUNIOR CLINICAL RESEARCH UNIT OVERVIEW The Breast Cancer Clinical Research Unit ( BCCRU ) focuses on the translational interface of therapeutic development. Breast cancer is a heterogeneous disease, and thus there are large inter-patient variations in terms of disease course, prognosis, relapse and resistance to conventional or targeted therapeutics. Our activities are directed towards personalised treatment, and range from preclinical models to the sponsoring of multicentric clinical trials. Specifically, our research areas are : ɗ Discovery of new targets for breast cancer prevention : role of fatty acid synthase ( FASN ). ɗ Breast cancer functional taxonomy : by using a systems biology approach, we are clustering the disease into subtypes defined by biologic features that constitute therapeutic targets. “ In 2016, the Breast Cancer Group has tackled the mechanisms of resistance against antiangiogenics, implementing these findings into clinical trials.” ɗ Study of the mechanisms of resistance against targeted therapies. ɗ Conduct investigator- initiated clinical trials. Miguel Quintela-Fandino Clinical Research Unit Head Staff Scientists María José Bueno, Silvana A. Mouron SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 123 CLINICAL RESEARCH PROGRAMME | BREAST CANCER JUNIOR CLINICAL RESEARCH UNIT RESEARCH HIGHLIGHTS In the field of functional taxonomy, we have completed our study in triple-negative breast cancer. We have interrogated the disease from the bimodal relapse pattern point of view, and performed a phosphoproteomic screening that would reduce the countless patterns of genomic, epigenomic and transcriptomic aberrations into a discrete number of patterns of hardwired signalling pathways. We found 6 kinases whose hyperactivity accounted for 94% of the relapsed cases. These kinases were grouped into a maximum number of 34 patterns, the largest of which ( 25%) was virtually associated with cure. This taxonomy was also useful because all the kinases in the final ‘ relapse signature ’ were also targetable nodes. Regarding the study of targeted therapies, we have observed that the generally assumed hypothesis of vascular normalisation upon exposure to antiangiogenics is not always true. In fact, resistance against antiangiogenics can originate after a vascular normalising or ‘ abnormalising ’ response. Whether a tumour experiences the former or the latter depends on the tumour type and the type of agent. What is quite important from the clinical point of view is that we can track, individually, whether a tumour experiences a normalising or an abnormalising response after less than 2 weeks of exposure to the agent, using a non-invasive imaging test with 18F-fluoromisonidazole. This has been demonstrated in animals and in patients. The applicability of this finding lies in the fact is that we have also unravelled the mechanisms of resistance depending on whether the tumour reacts with normalisation or abnormalisation against antiangiogenics : in the first case, the tumour switches from glycolytic to mitochondrial metabolism, which is reversible by mitochondrial inhibitors. In the latter, the tumour experiences an immune-switch. Since both mechanisms are targetable, we can now individually track which pathway a tumour is undergoing upon exposure to antiangiogenics and tailor which synergistic agent that patient would need. s Clinical Research Fellow Laura M. Medina ( until July ) Post-Doctoral Fellow Franciso J. Blanco ( until June ) Graduate Students Sara Fernández, Gonzalo Pérez, Ivana Zagorac Technicians Verónica Jiménez, José Francisco López ( TS )*, Esperanza Martín *Titulado Superior ( Advanced Degree ) ∞ PUBLICATIONS ∞ Quintela-Fandino M, Lluch A, Manso LM, Calvo I, Cortes J, Garcia Saenz JA, Gil JM, Martinez-Jez N, Gonzalez Martin A, Adrov- er E, De Andres R, Viñas G, Llombart-Cuss- ac A, Alba E, Guerra K, Bermejo B, Zamora E, Moreno-Anton F, Pernas Simón S, Car- rato A, Lopez A, Escudero MJ, Campo R, Carrasco EM, Palacios J, Mulero F, Colomer R ( 2016 ). 18F-fluoromisonidazole PET and activity of neoadjuvant nintedanib in early HER2-negative breast cancer : a window-of-opportunity randomized trial. Clin Cancer Res. PMID : 27587436. ∞ Mateo F et al. ( incl. Quintela-Fandino M ) ( 2016 ). Stem cell-like transcriptional reprogramming mediates metastatic re- sistance to mTOR inhibition. Oncogene. PMID : 27991928. ∞ Navarro P, Bueno MJ, Zagorac I, Mondejar T, Sanchez J, Mouron S, Muñoz J, Gomez- Lopez G, Jimenez-Renard V, Mulero F, Chandel NS, Quintela-Fandino M ( 2016 ). Targeting tumor mitochondrial metabo- lism overcomes resistance to antiangio- genics. Cell Rep 15, 2705-2718. ∞ Manso L, Mouron S, Tress M, Gomez-Lopez G, Morente M, Ciruelos E, Rubio-Camaril- lo M, Rodriguez-Peralto JL, Pujana MA, Pisano DG, Quintela-Fandino M ( 2016 ). Analysis of paired primary-metastatic hormone-receptor positive breast tum- ors ( HRPBC ) uncovers potential novel drivers of hormonal resistance. PLoS One 11, e0155840. ∞ Bueno M, Sanchez J, Colomer R, Quin- tela-Fandino M ( 2016 ). Antiangiogenics and hypoxic response : role of fatty acid synthase inhibitors. Curr Drug Targets 17, 1735-1746. ∞ Ebos J, Quintela-Fandino M ( 2016 ). Re- visiting the revolution : Examining the evolving role of antiangiogenic therapy in cancer. Curr Drug Targets 17, 1706. ∞ Lord S, Funes JM, Harris AL, Quintela-Fan- dino M ( 2016 ). Antiangiogenic resistance and cancer metabolism : opportunities for synthetic lethality. Curr Drug Targets 17, 1714-1727. ∞ PATENT ∞ Gold D.P., Quintela-Fandiño M.A. ( 2016 ). Combination Therapies. WO/2016/126618. ∞ AWARDS AND RECOGNITION ∞ 2016 AstraZeneca Award for Young In- vestigators, Spain. Figure Depiction of the metabolic adaptation of tumours when experiencing vascular normalisation upon exposure to antiangiogenics. An alternative response, increased vascular abnormality, occurs in roughly 30% of the cases. This response is coupled with immune reprogramming. ANNUAL REPORT 2016 124 VICE-DIRECTION OF TRANSLATIONAL RESEARCH PROSTATE CANCER JUNIOR CLINICAL RESEARCH UNIT OVERVIEW Prostate cancer (PrCa) is the most common cancer and the 2nd leading cause of cancer mortality among men in Western countries. Despite advances in diagnosis and early-disease treatment, up to 30% of PrCa patients will develop metastasis at some point and succumb after the acquisition of a castration-resistant status (mCRPrCa). The early identification of PrCa patients with greater predisposition to develop aggressive mCRPC could lead to the development of novel treatment strategies and improved outcomes. In addition to AR aberrations following androgen-deprivation therapy leading to resistance to current treatment options, DNA repair defects have been identified in about 5% and 25% of early PrCa and mCRPC, respectively. Seminal work from our Group, and others, has established that some alterations, e.g. germline BRCA1/BRCA2 deleterious mutations, are linked to poor outcomes. Currently, we lack the adequate biological knowledge and reliable biomarkers to select the right treatment for the right patient at the right time. RESEARCH HIGHLIGHTS PROCURE biomarkers platform This network was started by our Group in 2013 ; it currently has 5 ongoing prospective studies ( PROREPAIR, PROSTAC, PROSABI, PROSENZA, PRORADIUM ) in mCRPC in 63 participating centres with over 900 enrolled patients. PROREPAIR study This is a prospective multicentre cohort study involving 50 Spanish centres within the PROCURE network. By April 2016, 432 mCRPC patients were enrolled to evaluate the prevalence and impact of DNA repair germline mutations in mCRPC survival and the response to systemic treatments for mCRPC. Germline mutations were analysed in the following genes : ATM, ATR, David Olmos Junior Clinical Research Unit Head Clinical Investigator Elena Castro Clinical Research Fellow Nuria Romero SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 125 CLINICAL RESEARCH PROGRAMME | PROSTATE CANCER JUNIOR CLINICAL RESEARCH UNIT BARD1, BRCA2, BRCA1, BRIP1, CHEK2, GEN1, MLH1, MRE11A, MSH2, MSH6, NBN, PALB2, PMS2, RAD51C, RAD51D and XRCC2. Current results suggest that up to 12% of the patients in this series harbour a germline deleterious mutation. Analyses of the clinical impact of germline and somatic mutations in outcomes are still undergoing. BRCARAD and BRCAPROS studies, although in a retrospective fashion, will address similar questions at an early prostate cancer stage. SWITCH Phase II study In 2016, we also completed the enrolment and follow-up of our first clinical trial, ‘ Phase II pilot study of the prednisone to dexamethasone switch in mCRPC patients with progression on abiraterone and prednisone ’, aimed at analysing the role of certain steroids in the resistance and response to novel androgen- synthesis inhibitors in 26 patients. A simple change in prednisone to dexamethasone rescued the sensitivity to abiraterone and prolonged the time benefiting from this treatment in 40% of the patients ; such responses could be linked to AR mutations detected in ctDNA. Biological characterisation of BRCA2 and ATM mutated tumours Initial results from human tumour characterisation and mouse models conducted by our Group support that BRCA2 germline and/or somatic alterations may occur early in cancer progression, and that ATM aberrations will favour cancer progression and early intratumour heterogeneity. s Graduate Students Ylenia Cendón, Lorena Magraner ( since April ), Paz Nombela, Floortje Van De Poll ( until May ) Technician Vanessa Cañadilla Student in practice Noemi Hernández ( since September ) Visiting scientists Teresa Garcés ( since February ), Gala Grau ( since June ), Ana M. Gutiérrez ( since May ), Fernando López, Maria I. Pacheco, Leticia Rivera ( since May ) ∞ PUBLICATIONS ∞ Lorente D et al. ( incl. Olmos D ) ( 2016 ). Decline in Circulating Tumor Cell Count and Treatment Outcome in Advanced Prostate Cancer. Eur Urol 70, 985-992. ∞ Mateo J et al. ( incl.Castro E, Olmos D ) ( 2016 ). DNA Repair in Prostate Cancer : Biology and Clinical Implications. Eur Urol. PMID : 27590317. ∞ Missiaglia E et al. ( incl. Olmos D ) ( 2016 ). MicroRNA and gene co-expression net- works characterize biological and clinical behavior of rhabdomyosarcomas. Cancer Lett. PMID : 27984116. ∞ Castro E et al. ( 2016 ). The PROFILE Fea- sibility Study : Targeted Screening of Men With a Family History of Prostate Cancer. Oncologist 21, 716-722. ∞ Mateo J et al. ( incl. Olmos D ) ( 2016 ). A first in man, dose-finding study of the mTORC1/mTORC2 inhibitor OSI-027 in patients with advanced solid malignan- cies. Br J Cancer 114, 889-896. ∞ Castro E et al. ( incl. Olmos D ) ( 2016 ). The Role of PARP Inhibition in the Treatment of Castration-Resistant Prostate Cancer. Cancer J 22, 353-356. ∞ Selfe J et al. ( incl. Olmos D ) ( 2016 ). Impact of fusion gene status versus histology on risk-stratification for rhabdomyosarcoma : Retrospective analyses of patients on UK tri- als. Pediatr Blood Cancer. PMID : 28035744. ∞ AWARDS AND RECOGNITION ∞ Research contract from the Ramón y Cajal Programme, Ministerio de Economía, In- dustria y Competitividad ( MEIC ). ∞ Scientific Committee Member, ESMO Congress, Copenhagen, Denmark. ∞ Faculty Board Member, EORTC-ECCO- AACR-ESMO Methods in Clinical Cancer Research Workshop, Zeist, Netherlands. ∞ Nuria Romero was awarded the ‘ Best Communication ’ Award, 2nd Androgen Project Meeting in Prostate Cancer, Spain. ∞ Elena Castro was the recipient of the Best ESMO Fellowship Project ( ESMO Congress, Denmark ) and the Juan de la Cierva Research Contract ( MEIC, Spain ). Figure Prostate Cancer associated to somatic BRCA2 biallelic loss. Heterogeneity and clonal diversity was established based on the frequency and distribution of different dominant events for Prostate Cancer by FISH, as well as targeted sequencing focused on primary prostate cancer and a CRPC bone metastasis. ANNUAL REPORT 2016 126 VICE-DIRECTION OF TRANSLATIONAL RESEARCH MOLECULAR DIAGNOSTICS UNIT OVERVIEW The Molecular Diagnostics Unit ( MDU ) is mainly dedicated to developing, implementing, standardising and making available a wide variety of highly sensitive and specific molecular diagnostics assays that are scarcely available in the Hospitals of the Spanish National Health System. MDU’s portfolio of genetic tests enables the determination of alterations in the sequence or expression levels of key genes involved in cancer. In turn, these assays can be used for the early diagnosis of neoplasias, the detection of minimal residual disease in patients showing clinical remission, for monitoring the response to therapy in patients, as well as for facilitating decision-making amongst different treatment options. Furthermore, the Unit also provides support to the research needs of CNIO’s Clinical Research Units and Research Groups by checking their samples for alterations in the biomarkers included in our portfolio. Finally, MDU is very much committed to disseminating knowledge in the field of molecular diagnostics by hosting and mentoring biomedical students. Luis Lombardía Unit Head Technician Diana Romero “ In this transition phase of precision medicine, MDU is increasingly focused on the implementation of assays for the detection of biomarker alterations that could grant a more selective diagnosis for cancer patients.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 127 CLINICAL RESEARCH PROGRAMME | MOLECULAR DIAGNOSTICS UNIT RESEARCH HIGHLIGHTS Strengthening our support During 2016, our catalogue has grown with the addition of a new molecular diagnostics test based on the detection, by bi-directional Sanger sequencing, of mutations in exons 4 and 5 of the MYD88 gene. Waldenström’s macroglobulinemia ( WM ) is a rare form of blood cancer that is characterised by an excess of malignant white blood cells ( lymphoplasmacytic cells ) in the bone marrow. It has been shown that WM is the result of a multistep transformation process that accumulates sequential oncogenic alterations. The most prominent is the L265P somatic activating mutation in the MYD88 gene ( present in 90% of WM ). Hence, its detection would enable us to differentiate WM ( but also diffuse large B-cell vitreoretinal lymphoma or marginal zone lymphomas ) from indolent B-cell or other chronic lymphoproliferative disorders. Additionally, because identification of several gene alterations involved in the onset of myeloproliferative neoplasms ( MPNs ) has revealed the huge complexity of these diseases and has challenged their accurate differential diagnosis, we started working on the implementation and validation of a new assay that will enable us to detect mutations in the TET2 gene ; this will complement the diagnosis of MPNs patients. Mutations in this tumour suppressor gene ( present in 13% of MPNs ) lead to genomic instability via epigenetic modifications and foster cancer progression. Recent studies have revealed that the order in which these mutations are acquired is critical. Thus, patients with early mutations in TET2 were more likely to have better prognosis compared to patients who had previous mutations in others genes linked to MPNs ( FIGURE ). Lastly, we have completed the initial experimental phase of a clinical trial sub project, FRAGANCE, led by the CNIO Gastrointestinal Cancer Clinical Research Unit, which is geared towards precision medicine for fragile patients with advanced pancreatic cancer. Tutoring MDU has also upheld its policy regarding training programmes in 2016 by welcoming one medical resident and one undergraduate student. s Figure The detection of mutations in TET2 will improve the diagnostics algorithm by allowing prediction of the prognosis of patients with MPNs ( MPN : myeloproliferative neoplasm ; CML : chronic myeloid leukaemia ; PV : polycythaemia vera ; ET : essential thrombocythaemia ; PMF : primary myelofibrosis ). ANNUAL REPORT 2016 128 VICE-DIRECTION OF TRANSLATIONAL RESEARCH H12O-CNIO HAEMATOLOGICAL MALIGNANCIES CLINICAL RESEARCH UNIT OVERVIEW The Haematological Malignancies Clinical Research Unit focuses on 3 main objectives : ɗ Molecular research of haematological cancer : the study of cancer- induced changes at the proteomic and genomic levels. We aim to : i ) find new genomic and proteomic biomarkers for a better diagnosis of these haematological diseases ; ii ) identify new molecular alterations as predictors of response to treatment, e.g. to study minimal residual disease ; and iii ) study immune mechanisms of cancer control, with a special focus on NK cells. ɗ In vitro research : i ) to establish the effects of new anticancer molecules in in vitro models of the disease ; ii ) to determine the mechanisms of resistance to anticancer drugs. ɗ Clinical research : to translate preclinical findings to the patients through a phase I clinical trials unit. Joaquín Martínez-López Clinical Research Unit Head “ We contribute towards redefining the response criteria for Multiple Myeloma ( MM ) through the usage of new molecular techniques. In 2016, we published the first reports sequencing complete exomes of amyloidosis. Finally, we reported a new cell therapy approach based on infusion of NK cells in MM.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 129 CLINICAL RESEARCH PROGRAMME | H12O-CNIO HAEMATOLOGICAL MALIGNANCIES CLINICAL RESEARCH UNIT RESEARCH HIGHLIGHTS The most relevant achievements of our Group in 2016 were : ɗ We reported a phase I clinical trial based on an innovative cell therapy approach using activated and expanded NK cells for Multiple Myeloma ( MM ). The results of exploring this approach in phase II and III clinical trials are promising. ɗ We published the first report of exome sequencing in amyloidosis. We have not found any recurrent mutation. ɗ Finally, we redefined the role of stringent complete response by next generation sequencing in Multiple Myeloma. s Staff Scientists Rosa Ayala, Teresa Cedena, Inmaculada Rapado, Beatriz Sánchez-Vega Post-Doctoral Fellows Lucía Fernández ( since July ), María Linares, Ricardo Sánchez, Antonio Valeri Graduate Students Alicia Arenas, Isabel Cuenca, Alejandra Leivas, M. Luz Morales ( since June ), Esther Onecha, Alejandra Ortíz ( since May ), Yanira Ruíz Technicians Alba García, Ana I. Sánchez ∞ PUBLICATIONS ∞ Fulciniti M, Martinez-López J, Senapedis W, Oliva S, Bandi RL, Amodio N, Xu Y, Szalat RL, Gulla A, Samur MK, Roccaro A, Linares M, Cea M, Baloglu E, Argueta C, Landesman Y, Shacham S, Liu S, Schenone M, Wu SL, Karger B, Prabhala R, Anderson KC, Munshi NC ( 2016 ). Functional role and therapeutic targeting of p21-associated kinase 4 ( PAK4 ) in multiple myeloma. Blood. PMID : 28096095. ∞ Cross NC et al. ( incl. Martinez-López J ) ( 2016 ). Development and evaluation of a secondary reference panel for BCR-ABL1 quantitation on the International Scale. Leukemia 30, 1844-1852. ∞ Santos-Lozano A, Morales-Gonzalez A, Sanchis-Gomar F, Cristi-Montero C, Fiuza-Luces C, Pareja-Galeano H, Martínez-López J, Garatachea N, Lucia A ( 2016 ). Response rate to the treatment of Waldenström macroglobulinemia : A meta-analysis of the results of clin- ical trials. Crit Rev Oncol Hematol 105, 118-126. ∞ SELECTED PUBLICATIONS AT OTHER INSTITUTIONS ∞ Martínez-Moreno M, Leiva M, Aguil- era-Montilla N, Sevilla-Movilla S1, de Val SI, Arellano-Sánchez N, Gutiérrez NC, Maldonado R, Martínez-López J, Buño I, García-Marco JA, Sánchez-Mateos P, Hidalgo A, García-Pardo A, Teixidó J ( 2016 ). In vivo adhesion of malignant B cells to bone marrow microvasculature is regulated by a4β1 cytoplasmic-binding proteins. Leukemia 30, 861-872. ∞ Paiva B, Puig N, Cedena MT, de Jong BG, Ruiz Y, Rapado I, Martinez-Lopez J, Cor- don L, Alignani D, Delgado JA, van Zelm MC, Van Dongen JJ, Pascual M, Aguirre X, Prosper F, Martín-Subero JI, Vidriales MB, Gutierrez NC, Hernandez MT, Oriol A, Echeveste MA, Gonzalez Y, Johnson SK, Epstein J, Barlogie B, Morgan GJ, Orfao A, Blade J, Mateos MV, Lahuerta JJ, Miguel JF ( 2016 ). Differentiation stage of mye- loma plasma cells : biological and clinical significance. Leukemia. PMID : 27479184. ∞ Gutiérrez-González A, Martínez-More- no M, Samaniego R, Arellano-Sánchez N, Salinas-Muñoz L, Relloso M, Valeri A, Martínez-López J, Corbí ÁL, Hidalgo A, García-Pardo Á, Teixidó J, Sánchez-Ma- teos P ( 2016 ). Evaluation of the potential therapeutic benefits of macrophage re- programming in multiple myeloma. Blood 128, 2241-2252. ∞ Mateos MV, Martínez-López J, Hernández MT, Ocio EM, Rosiñol L, Martínez R, Teruel AI, Gutiérrez NC, Martín Ramos ML, Ori- ol A, Bargay J, Bengoechea E, González Y, Pérez de Oteyza J, Gironella M, Encinas C, Martín J, Cabrera C, Paiva B, Cedena MT, Puig N, Bladé J, Lahuerta JJ, San- Miguel J ( 2016 ). Sequential versus alter- nating administration of VMP and Rd in elderly patients with newly diagnosed MM. Blood 127, 420-425. ∞ Paiva B, Martinez-Lopez J, Corchete LA, Sanchez-Vega B, Rapado I, Puig N, Barrio S, Sanchez ML, Alignani D, Lasa M, García de Coca A, Pardal E, Oriol A, Gonzalez Garcia ME, Escalante F, González-López TJ, Palomera L, Alonso J, Prosper F, Or- fao A, Vidriales MB, Mateos MV, Lahuerta JJ, Gutierrez NC, San Miguel JF ( 2016 ). Phenotypic, transcriptomic and genomic features of clonal plasma cells in light chain amyloidosis. Blood 127, 3035-3039. ∞ Paiva B, Cedena MT, Puig N, Arana P, Vidriales MB, Cordon L, Flores-Montero J, Gutierrez NC, Martín-Ramos ML, Martin- ez-Lopez J, Ocio EM, Hernandez MT, Teruel AI, Rosiñol L, Echeveste MA, Martinez R, Gironella M, Oriol A, Cabrera C, Martin J, Bargay J, Encinas C, Gonzalez Y, Van Dongen JJ, Orfao A, Bladé J, Mateos MV, Lahuerta JJ, San Miguel JF ( 2016 ). Minimal residual disease monitoring and immune profiling using second generation flow cytometry in elderly multiple myeloma. Blood 127, 3165-3174. Figure Phenotyping of multiple myeloma macrophages ( MM-MØ ) from Bone Marrow ( BM ) patient samples. ( A ) Multi-coloured staining of BM aspirates containing particles from active disease MM patients, as indicated. Upper panels represent panoramic views, whereas bottom panels are magnified ones. Nuclear- 49,6-diamidino-2-phenylindole appears in blue in all cases. ( B ) Plot showing the mean fluorescence intensity for each marker in CD1631 tumour associated macrophages ( TAM ; n = 10 cases ). Cells > 25 arbitrary units ( a.u.) are considered positive, relative to negative control. Scale bars as indicated. ANNUAL REPORT 2016 130 VICE-DIRECTION OF TRANSLATIONAL RESEARCH H12O-CNIO LUNG CANCER CLINICAL RESEARCH UNIT OVERVIEW Our Group combines basic preclinical studies with clinical and translational research, mainly in lung cancer and other solid tumours. In summary, the main research areas of our Group focus on 2 modalities : ( 1 ) the identification of new molecular biomarkers that can be used in the clinic for diagnostic, prognostic, predictive and pharmacogenomic purposes ; and ( 2 ) developing novel treatment strategies. For example, we have comprehensively profiled bronchoalveloar lavage ( BLA ) fluids of COPD and lung cancer patients, showing a differential mi-RNA, protein and inflammatory cytokine expression between both diseases and different subtypes of lung cancer. On the other hand, we have developed a patient-derived xenograft ( PDX ) platform of non-small-cell lung cancers to test new drugs/targets. We are also developing PDXs of small-cell lung cancers. Finally, our Group has extensive experience in the development of new drugs, as well as in conducting practice-changing phase II/III trials in the fields of precision oncology and immunooncology. Luis G. Paz-Ares Clinical Research Unit Head Staff Scientists M. Teresa Argullo, Daniel E. Castellano, Irene Ferrer, Rocio García, Blanca Homet ( until September ), Lara C. Iglesias, Sonia Molina, Santiago Ponce, Jon Zugazagoitia “ Our Group has significantly contributed to the discovery of biomarkers as well as to the early development of new drugs tailoring novel targets. We have co-led randomised clinical trials with biological therapies and immunotherapy in lung cancer and other solid tumours.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 131 CLINICAL RESEARCH PROGRAMME | H12O-CNIO LUNG CANCER CLINICAL RESEARCH UNIT RESEARCH HIGHLIGHTS New drug development and early clinical trials Our Group has been actively involved in pharmacogenomic, pharmacokinetic, translational and clinical studies with novel antitumour agents in several types of solid tumours, particularly lung cancer. Our principal clinical research area has been immunotherapy and immune-based early clinical trials. As a first relevant example we can mention the CheckMate CA 209-032 trial testing nivolumab +/- ipilimumab in recurrent or extensive-stage small-cell lung cancer, which was fully recruited in 2016, with a substantial contribution by investigators from our Group. These important data have been recently published in The Lancet Oncology. In addition, Luis Paz-Ares is the principal investigator of a phase I trial ( JDVF ) testing a novel combination of pembrolizumab plus ramurirumab in different types of solid tumours. Encouraging preliminary clinical data were presented at ASCO 2016 in the cohort of non-small-cell lung cancer, showing a response rate of 35 % and 7-months of progression-free survival in pretreated patients. Finally, a first-in-human trial with a novel T-cell bispecific antibody targeting carcinoembryonic antigen ( CEA ) expressed on tumour cells and CD3 on T-cells was initiated and is actively recruiting patients. Conducting practice changing randomised controlled trials Our Group has also made a substantial contribution in conducting pivotal trials with immune checkpoint inhibitors. In particular, an important phase III trial, led by Dr Paz-Ares ( the international principal investigator ), with pembrolizumab in completely resected non-small cell lung cancer patients is actively recruiting participants. Furthermore, the first randomised trial comparing second-generation ( afatinib ) versus first generation ( gefitinib ) tyrosine-kinase inhibitors in patients with EGFR-mutant lung cancers, also internationally led by Dr Paz Ares, was completed in 2016 and its results were recently published in The Lancet Oncology. Novel biomarker development and translation IL-11 and CCL-1 have been proposed as novel diagnostic biomarkers of lung adenocarcinoma in bronchoalveolar lavage fluid. This finding has potential implications in early lung cancer diagnosis. Moreover, different members of our Group contributed towards providing further insights into the role of PD-L1 expression and other potential immune biomarkers for the benefit of immune checkpoint inhibitors. s Post-Doctoral Fellow Beatriz Soldevilla ( since June ) Graduate Students Ángela Marrugal, Laura Ojeda, Álvaro Quintanal, Patricia Yagüe ( since October ) Technicians Laura García, Rocío Suárez, M. José Durán ( since July ), Virginia Pardo ( since February ) ∞ PUBLICATIONS ∞ Hirsch FR, Scagliotti GV, Mulshine JL, Kwon R, Curran WJ Jr, Wu YL, Paz-Ares L ( 2016 ). Lung cancer : current therapies and new targeted treatments. Lancet. PMID : 27574741. ∞ Park K, Tan EH, O’Byrne K, Zhang L, Boyer M, Mok T, Hirsh V, Yang JC, Lee KH, Lu S, Shi Y, Kim SW, Laskin J, Kim DW, Arvis CD, Kölbeck K, Laurie SA, Tsai CM, Sha- hidi M, Kim M, Massey D, Zazulina V, Paz- Ares L ( 2016 ). Afatinib versus gefitinib as first-line treatment of patients with EGFR mutation-positive non-small-cell lung cancer ( LUX-Lung 7 ): a phase 2B, open-label, randomised controlled trial. Lancet Oncol 17, 577-589. ∞ Zugazagoitia J, Ferrer I, Paz-Ares L ( 2016 ). Osimertinib in EGFR-mutant NSCLC : how to select patients and when to treat. Lan- cet Oncol 17, 1622-1623. ∞ Spigel DR et al. ( incl. Paz-Ares L ) ( 2016 ). Results From the Phase III Randomized Trial of Onartuzumab Plus Erlotinib Versus Erlotinib in Previously Treated Stage IIIB or IV Non-Small-Cell Lung Cancer : METLung. J Clin Oncol. PMID : 27937096. ∞ Paz-Ares L et al. ( 2016 ). Correlation of EGFR-expression with safety and effica- cy outcomes in SQUIRE : a randomized, multicenter, open-label, phase III study of gemcitabine-cisplatin plus necitumumab versus gemcitabine-cisplatin alone in the first-line treatment of patients with stage IV squamous non-small-cell lung cancer. Ann Oncol 27, 1573-1579. ∞ Ramalingam SS, O’Byrne K, Boyer M, Mok T, Jänne PA, Zhang H, Liang J, Taylor I, Sbar EI, Paz-Ares L ( 2016 ). Dacomitinib versus erlotinib in patients with EGFR-mu- tated advanced nonsmall-cell lung cancer ( NSCLC ): pooled subset analyses from two randomized trials. Ann Oncol 27, 1363. ∞ Molina-Pinelo S, Salinas A, Moreno-Ma- ta N, Ferrer I, Suarez R, Andrés-León E, Rodríguez-Paredes M, Gutekunst J, Jantus-Lewintre E, Camps C, Carnero A, Paz-Ares L ( 2016 ). Impact of DLK1-DIO3 imprinted cluster hypomethylation in smoker patients with lung cancer. Onco- target. PMID : 27447969. ∞ Usó M et al. ( incl. Molina-Pinelo S, Paz- Ares L ) ( 2016 ). Analysis of the immune microenvironment in resected non-small cell lung cancer : the prognostic value of different T lymphocyte markers. Onco- target 7, 52849-52861. ∞ Liu  x  et al . (   incl. Castellano D, Rodríguez-Antona C ) ( 2016 ). Meta-anal- ysis on the association of VEGFR1 ge- netic variants with sunitinib outcome in metastatic renal cell carcinoma patients. Oncotarget. PMID : 27901483. ∞ Diekstra MH et al. ( incl. Castellano D, Rodríguez-Antona C ) ( 2016 ). Sunitinib in- duced hypertension in CYP3A4 rs4646437 A-allele carriers with metastatic renal cell carcinoma. Pharmacogenomics J. PMID : 26810136. ∞ Zugazagoitia J, Guedes C, Ponce S, Fer- rer I, Molina-Pinelo S, Paz-Ares L ( 2016 ). Current Challenges in Cancer Treatment. Clin Ther 38, 1551-1566. ∞ Marrugal Á, Ojeda L, Paz-Ares L, Moli- na-Pinelo S, Ferrer I ( 2016 ). Proteom- ic-Based Approaches for the Study of Cytokines in Lung Cancer. Dis Markers 2016, 2138627. ∞ Quintanal-Villalonga A, Paz-Ares L, Ferrer I, Molina-Pinelo S ( 2016 ). Tyrosine Ki- nase Receptor Landscape in Lung Cancer : Therapeutical Implications. Dis Markers 2016, 9214056. ANNUAL REPORT 2016 132 VICE-DIRECTION OF TRANSLATIONAL RESEARCH BIOBANK OVERVIEW The CNIO Biobank is a cross-service platform for CNIO researchers, as well as the general scientific community, and is geared towards the promotion of biomedical research in cancer and related diseases. The CNIO Biobank facilitates access to human samples for researchers, ensuring that both the acquisition and use of human samples complies with all the legal and ethical principles that protect donors ’ rights. The CNIO Biobank is a ‘ biobank for biomedical research purposes ’, as defined by the Spanish Law 14/2007 on Biomedical Research and the Royal Decree RD 1716/2011. It is therefore defined as a public, non-profit organisation that hosts several collections of human biological samples for biomedical research. The biobank is organised as a technical unit with strict criteria for quality, order and purpose, regardless of whether or not it hosts other collections of biological samples for different purposes. Samples and their associated information are managed in compliance with Spanish legislation and international recommendations ; all of this is consistent with quality criteria for sample collection and its subsequent management. The Biobank has been authorised by the Health Authorities of the Comunidad Autónoma de Madrid – in accordance with the regulation established by RD1716/2011 – and is registered in the National Registry of Biobanks with reference B.000848. Manuel M. Morente Director Technicians Nuria Ajenjo ( TS )*, Inmaculada Almenara, M. Jesús Artiga ( TS ), Francisco De Luna ( TS ) *Titulado Superior ( Advanced Degree ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 133 CLINICAL RESEARCH PROGRAMME | BIOBANk RESEARCH HIGHLIGHTS Biobanking ɗ Collection, management, manipulation and custody of human biological samples and associated documentation, in accordance with the legal framework for biobanking. ɗ Transfer of samples and clinical biomedical information to research projects, under the approval of the corresponding scientific and ethical committees. Management of other collections ɗ Custody service of collections of biological samples and/or information related to biomedical research as promoted by the CNIO or other external research groups. ɗ Coordination of sample collections in multicentre studies. ɗ Processing of products derived from human samples for research ( tissue arrays, DNA, RNA, etc.). ɗ Researchers who want to deposit their collections at the CNIO- Biobank facilities, or who wish to request samples, must sign an MTA ( Material Transfer Agreement ) that specifies the terms and conditions under which the Biobank will custody the samples and data. Ethico-legal advice for CNIO researchers regarding the use of human samples in biomedical research ɗ Technical, scientific and ethical advice regarding the collection, storage and management of human samples used for biomedical research, as well as in regards to the creation and management of new collections that are beyond the Biobank’s scope. Other services ɗ Collaboration with CNIO researchers in human pathology. ɗ Collaboration in diagnostic activities as specialists in human pathology. During 2016, the CNIO-Biobank has supported 9 tissue requests from scientific research projects with 180 single cases and 303 tissue microarrays transferred. Additionally, as the Spanish National Biobank Network Coordination Office, we have managed 37 scientific research projects of high complexity. The mean impact factor of the 10 publications published in 2016, for which our Unit provided support was 11.384. We also provided sample and/or documental support for the familial cancer activities of the CNIO Human Cancer Genetics Programme. The CNIO Biobank participates in and coordinates the Spanish National Biobank Network. This nationwide platform of services integrates 52 institutions ( www.redbiobancos.es ) and is an initiative of the Instituto de Salud Carlos III ( ISCIII ). Finally, the Unit has spearheaded many activities in the national and international biobanking scene through its participation and leadership in numerous forums, working groups and national and international scientific societies. These include the European, Middle Eastern and African Society for Biopreservation and Biobanking ( ESBB ), the International Society for Biological and Environmental Repositories ( ISBER ), international think tanks such as the Marble Arch International Working Group on Clinical Biobanking, BC-Net IARC-WHO/NCI initiative, EurocanPlatform ( 7th FP ), and others. s ∞ PUBLICATIONS ∞ Manso L et al. ( incl. Mourón S, Tress M, Gómez-López G, Morente M, Ciruelos E, Rubio-Camarillo M, Pisano DG, Quinte- la-Fandino M ) ( 2016 ). Analysis of Paired Primary-Metastatic Hormone-Receptor Positive Breast Tumors ( HRPBC ) Uncovers Potential Novel Drivers of Hormonal Re- sistance. PLoS One 11, e0155840. ∞ Doucet M et al. ( incl. Morente M ) ( 2016 ). Quality Matters : 2016 Annual Conference of the National Infrastructures for Biobank- ing. Biopreserv Biobank. PMID : 27992240. ∞ Suárez AE et al. ( incl. Artiga MJ ) ( 2016 ). Angioimmunoblastic T-cell lymphoma with a clonal plasma cell proliferation that underwent immunoglobulin iso- type switch in the skin, coinciding with cutaneous disease progression. J Cutan Pathol 43, 1203-1210. ∞ García-Donas J et al. ( incl. Inglada-Pérez L, Graña O, Apellániz-Ruiz M, Morente M, Pisano DG, Robledo M, Rodriguez-Anto- na C ) ( 2016 ). Deep sequencing reveals microRNAs predictive of antiangiogenic drug response. JCI Insight 1, e86051. ∞ AWARDS AND RECOGNITION ∞ Member, Evaluation panel ‘ Enabling Ger- man Biobank Sites to Connect to BBM- RI-ERIC ’, Federal Ministry of Education and Research, Germany. Figure Biobanking is a transversal activity based on four basic commitments : respect for donors’ rights, technical excellence, adherence to the scientific requirements, and strict compliance with Spanish legislation. ANNUAL REPORT 2016 134 Direction of Innovation SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 135 Biotechnology Programme 138 Genomics Core Unit 140 Transgenic Mice Core Unit 142 Monoclonal Antibodies Core Unit 144 Molecular Imaging Core Unit 146 Flow Cytometry Core Unit 148 Confocal Microscopy Core Unit 150 Proteomics Core Unit 152 Histopathology Core Unit 154 Animal Facility 156 Experimental Therapeutics Programme 158 Medicinal Chemistry Section 160 Biology Section 164 CNIO - Lilly Cell Signalling Therapies Section 168 CNIO - Lilly Epigenetics Section 170 Technology Transfer and Valorisation Office 172 Private Sponsors 174 ANNUAL REPORT 2016 136 ÓSCAR FERNÁNDEZ-CAPETILLO Director of Innovation SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 137 “ Innovation and research constitute both sides of the same coin. At the CNIO, we are making our effort so that the products of our research end up benefiting society.” Scientific research often brings about the generation of knowledge and/or products that can be of use beyond the boundaries of academic research. These can include novel technologies, animal models, antibodies or chemical entities with interesting biomedical properties, among others. At the CNIO, we aim to bridge the gap between our researchers and potential outsources that could potentially be interested in further developing our inventions. These initiatives have materialised in the form of contracts with industry or in the licensing out of several of our products. Importantly, while the royalties deriving from these activities mostly benefit the CNIO as an Institution, they also circle back to the scientific Groups to fund their research as well as to the inventors themselves. To date, more than 40 investigators from 11 groups have benefited from this initiative. A Programme that deserves to be singled out in terms of innovation is that of the Experimental Therapeutics Programme ( ETP ), a whole department that aims to develop chemical entities that could potentially lead to new anticancer therapies. With several of these molecules already licensed out to the Pharmaceutical industry, their current portfolio of projects includes several at an advanced stage. The Direction of Innovation promotes a pipeline based on collaborative drug development between ETP and the rest of the CNIO Groups, so that drug-development capitalises on the excellent research conducted by our basic scientists. Accordingly, all of our current early stage drug development projects have emerged from active collaborations with scientific research groups. In addition, we are trying to consolidate ETP as a strong node for anticancer drug development in Spain, so that our expertise can contribute towards the development of new therapies together with other cancer researchers in our country. An important strength of our innovation activities derives from the excellent support provided by the Biotechnology Programme. Their work is not only vital for the progress of our scientific projects, but has also led to the realisation of important products and technologies that have contributed to our innovation portfolio. Some of the antibodies developed at the CNIO are contributing towards important aspects of current innovative treatments, such as those based on immunotherapies. The accreditation of our Animal Facility by the Association for Assessment and Accreditation of Laboratory Animal Care International ( AAALAC ) also represents a key milestone in 2016 for the Biotechnology Programme. Finally, several initiatives continue to be in place with the aim of fostering an innovation culture among our scientists, such as our continuous agreement with the prestigious IE Business School, through which many of our investigators have already obtained training in market-oriented innovation strategies. ANNUAL REPORT 2016 138 DIRECTION OF INNOVATION BIOTECHNOLOGY PROGRAMME FERNANDO PELÁEZ Programme Director SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 139 BIOTECHNOLOGY PROGRAMME The main mission of the Biotechnology Programme is to provide expert technical support and advice to CNIO Research Groups in a number of disciplines and technologies widely used in biomedical research, as well as to implement and develop state-of-the-art biotechnological tools and reagents for cancer research. The Programme is currently composed of nine Core Units covering major areas in Biotechnology, namely, Genomics, Proteomics, Monoclonal Antibodies, Histopathology, Flow Cytometry, Confocal Microscopy, Molecular Imaging and Transgenic Mice, as well as an Animal Facility. Although the Core Units are mainly focused on meeting the internal demand and collaborating with the CNIO Research Groups, they also provide support and collaborate with groups from other public institutions, as well as with private companies. In 2016, the Programme was significantly reinforced with the recruitment of 9 young technicians who are funded for a 2-year period by the programme Ayudas para la Promoción de Empleo Joven e Implantación de la Garantía Juvenil en I+D+i del Ministerio de Economía y Competitividad. This year, the CNIO Animal Facility obtained full accreditation from the Association for Assessment and Accreditation of Laboratory Animal Care International ( AAALAC ); this recognition reflects the high standards achieved by the CNIO with regards to the use and care of animals for experimentation. Faithful to its mission, a number of different technological innovations have been explored or implemented by the Core Units during this last year, often in collaboration with CNIO Groups. Noteworthy examples include the application of CRISPR technology for the generation of mouse models, the generation of monoclonal antibodies against small molecules, the application of proteomic approaches to the study of exosomes, the development of immuno- PET approaches for tumour imaging, the expansion of multicolour capabilities in flow cytometry studies, and the application of microfluidics-based setups to advanced microscopy, to name a few. In 2016, the Programme and its Core Units have been particularly active in networking activities. This included the participation of several of our Programme members in the Core Technologies for Life Sciences ( CTLS ) meeting at EMBL in Heidelberg, Germany, where the first steps were taken towards the organisation of a new scientific association addressing core facilities issues ( CTLS ). In addition, several Unit Heads were very active in participating in networks and scientific societies from their corresponding fields. Also, the Programme Director was voted as President-Elect of the Spanish Society of Biotechnology ( SEBiot ), highlighting the prominent role of the CNIO in this area. Also, as an indication of our high commitment to training and education, the Programme has been involved in the organisation of courses, workshops and specialised meetings. Moreover, an increasing number of our staff members undertook Masters and other training activities, at the CNIO and elsewhere. This year, the Core Units were particularly successful in attracting funding from external sources through activities related to innovation ; several contracts and agreements with private companies and public institutions, based on the technologies mastered by several of our Core Units, were formalised. Also, the royalties derived from the sales of the antibodies produced by the Monoclonal Antibodies Unit have grown by about 16% over the previous year, reaching a new historical maximum. Last but not least, 2016 has once again been a very productive year, scientifically, for the Programme. The contribution of the Units to the overall scientific performance of the CNIO is reflected in nearly 30 publications, many of them appearing in top journals. “ The best possible partner for enabling CNIO’s scientists to achieve their research goals is a strong Biotechnology Programme with state-of-the-art Core Facilities.” ANNUAL REPORT 2016 140 DIRECTION OF INNOVATION GENOMICS CORE UNIT “ The Genomics Unit, with its toolbox for DNA and RNA analyses, helps CNIO scientists to understand the molecular processes underlying cancer in a large number of basic and applied research projects.” Orlando Domínguez Core Unit Head Technicians Purificación Arribas, Guadalupe Luengo, Jorge Monsech, Ana Belén Moreno ( PEJ )*, Ángeles Rubio, Delfina Sanguino *Plan de Empleo Joven ( Youth Employment Plan ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 141 BIOTECHNOLOGY PROGRAMME | GENOMICS CORE UNIT OVERVIEW Genomics is the discipline that studies the structure and dynamics of the genome, its features, its regulation and expression. The genome is the core of life, the ensemble of the genetic material that conserves the assembly instructions of the species. Each cell in an individual keeps a copy of it deep in its nucleus. Chemically made of linear DNA macromolecules and distributed into chromosomes, it is packed with and interpreted by a myriad of protein cohorts acting in concert. It is expressed into RNA transcripts ; some are functional in and of themselves, and others, constitute an intermediate step leading to the functional proteins that govern the cell. While less than a 2% fraction of a mammalian genome codes for protein, a vast majority of it ( 80%) has been found to participate in biochemical events. The genome is not immutable, it can suffer alterations. A chance for biological evolution or for damage. In fact, cancer derives from the accumulation of such alterations. Cells with a damaged genome can transform and develop into a tumour. The field of Genomics sheds light on this world of complexity. RESEARCH HIGHLIGHTS All tumours, even those of the same type and sharing a similar fate, are molecularly different and heterogeneous at the molecular level. By employing a distinct set of powerful methodologies, Genomics reveals the genetic diversity of cancer and helps to dissect molecular mechanisms. These methodologies have the capacity to interrogate a wide number of genetic loci, or even a whole genome in a single assay. Some tools detect modifications at a structural level : mutations, binding of protein factors, variations in chromatin folding. Others are suitable for observing functional choreographies, transcriptomic changes – for example, in response to treatments – that may uncover therapeutic targets and prognostic biomarkers. The Genomics Unit provides services at two levels of complexity. The genomic wide level is addressed by both deep-sequencing and microarray technologies. Deep-sequencing permits a variety of applications, such as whole-genome or whole-exome tumour sequencing, transcriptome analyses by RNAseq, or location of interacting protein factors on chromosomal DNA by ChIPseq. As a novel sample type, the Unit successfully participated in the exome sequencing of cell free DNA obtained from cancer patients ’ blood. This year has seen a 40% increase in the overall demand and in the number of samples processed. On the other hand, the DNA microarray platform can be efficiently used for transcriptome determinations or for the detection of chromosomal copy number abnormalities. At the single locus level other offers are available. A traditional DNA capillary sequencing service, based on a 3730xl DNA Analyzer from Applied Biosystems, is being used to find and confirm mutations in candidate genes as well as for the verification of cloned genes or inserts. The Unit also provides a transgenic mouse genotyping service, based on allele-specific quantitative PCR for a quick and efficient turnaround time. With a current, but continuously growing, catalogue of over 30 genetic modifications, the demand for genotyping services this year has almost doubled in comparison to former years. s Figure An RNAseq experiment reveals the effects of a treatment at the RNA level. A blind analysis properly clusters samples of the same type together in columns. Genes whose transcription is significantly affected by treatment are shown in rows. Colour intensity reflects the transcriptional level. Some gene expression variability is apparent among replicates from the same condition. ANNUAL REPORT 2016 142 DIRECTION OF INNOVATION Sagrario Ortega Core Unit Head Student in Practice Aleida Pujol OVERVIEW Genetically engineered mice are an essential tool for analysing the molecular mechanisms underlying tumour development and cancer biology. Modelling cancer by modifying the germ line of the mouse has become a crucial component of drug discovery as well as for the assessment of experimental therapies at the preclinical stage. The Transgenic Mice Unit at the CNIO offers state-of-the- art technology for the manipulation of the mouse genome. Using classical transgenesis, homologous recombination in embryonic stem cells and genome editing by targeted nucleases, the Unit has generated more than 300 mutant alleles of cancer related genes in the mouse germ line. The Unit also provides support and collaborates with CNIO researchers in many aspects related to research with embryonic stem ( ES ) cells, induced pluripotent stem ( iPS ) cells, and embryo- and mouse model-based research. Finally, the Unit also leads its own research projects focused on the generation of mouse models to study tumour biology, as well as on the screening of cancer-related genes. TRANSGENIC MICE CORE UNIT “ In 2016, the Unit generated over a dozen GEM strains containing knockout and knockin mutations, using the CRISPR/Cas9 system of S. Pyogenes. The Unit contributed to 8 peer-reviewed articles, in collaboration with CNIO and external groups, including the description of a new mouse strain for conditional gene targeting of the lymphatic system.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 143 BIOTECHNOLOGY PROGRAMME | TRANSGENIC MICE CORE UNIT RESEARCH HIGHLIGHTS The CNIO Transgenic Mice Unit is dedicated to the generation, cryopreservation and derivation of genetically engineered mouse strains. We have created over 200 mutant strains, including knockout, knockin and conditional alleles, by gene targeting in embryonic stem ( ES ) cells, and over 100 mouse strains by conventional transgenesis. The Unit currently maintains a cryopreserved stock of over 1000 mouse strains, frozen at the Unit as sperm or embryos. This stock represents an invaluable resource of engineered strains for modelling and studying cancer in the mouse. Through our Unit, the CNIO shares part of this stock with EMMA ( the European Mouse Mutant Archive ) in order to make these models more accessible to the wider scientific community. We acknowledge the CNIO Animal Facility for their constant help and collaboration to make all these achievements possible. The CRISPR/Cas9 system of Streptococcus pyogenes has expanded the currently available set of mammalian genome engineering tools, providing an easy, efficient, flexible and versatile method for creating targeted mutations in mammalian genes. We use the CRISPR/Cas9 system to generate knockout and knockin mice by introducing the components of the system, the guide CRISPR RNA and the Cas9 nuclease ( either as messenger RNA or as protein ) directly into mouse zygotes ( FIGURE ). In our experience, this system has proven to be extremely efficient for introducing new additional mutations in strains that are already carrying several engineered alleles, such as some mouse models of lung and pancreatic cancer that are used at the CNIO. We have also used the system to generate knockin alleles ( point mutations ) and tag insertions with efficiencies close to 20% directly in zygotes. The characteristics of the CRISPR system − efficient, fast and easy to implement − make it extremely useful for creating constitutive mutations in the mouse and to test certain biological questions before embarking on the creation of conditional or more sophisticated alleles. For these types of alleles, gene targeting in ES cells may still be the method of choice and we are currently optimising the use of CRISPR in ES cells to increase the efficiency of this technology. s Technicians Estefania Ayala, M. Carmen Gómez, Jaime A. Muñoz ( TS )*, Lucía M. Pérez De Ayala ( PEJ-L )**, Patricia Prieto ( TS )*, Pierfrancesco Vargiu ( TS )* *Titulado Superior ( Advanced Degree ) **Plan de Empleo Joven-Licenciado ( Youth Employment Plan-Graduate ) F i g u re   Ef f i c i ency o f GFP knockout via CRISPR in mouse embryos. Embryos are collected from B6.CBA females, crossed with 129Gt( ROSA )26Sortm( CAG-EGFP ) Luo ( KI/KI ) males, at E0.5. Embryos are injected with gRNA_GFP97 ( 50ng/µl ) and commercial Cas9 protein ( 100ng/µl ) in the cytoplasm at the zygote stage and cultured in vitro for 3 days up to the blastocyst stage. Confocal images ( maximal projection ) of GFP fluorescence. ∞ PUBLICATIONS ∞ Ruiz S, Mayor-Ruiz C, Lafarga V, Murga M, Vega-Sendino M, Ortega S, Fernan- dez-Capetillo O ( 2016 ). A Genome-wide CRISPR Screen Identifies CDC25A as a Determinant of Sensitivity to ATR Inhib- itors. Mol Cell 62, 307-313. ∞ Ahuja AK, Jodkowska K, Teloni F, Bizard AH, Zellweger R, Herrador R, Ortega S, Hickson ID, Altmeyer M, Mendez J, Lopes M ( 2016 ). A short G1 phase imposes constitutive replication stress and fork remodelling in mouse embryonic stem cells. Nat Commun 7, 10660. ∞ Varela E, Muñoz-Lorente MA, Tejera AM, Ortega S, Blasco MA ( 2016 ). Generation of mice with longer and better preserved telomeres in the absence of genetic ma- nipulations. Nat Commun 7, 11739. ∞ Calvete O, Varro A, Pritchard DM, Barroso A, Oteo M, Morcillo MÁ, Vargiu P, Dodd S, Garcia M, Reyes J, Ortega S, Benitez J ( 2016 ). A knockin mouse model for hu- man ATP4aR703C mutation identified in familial gastric neuroendocrine tumors recapitulates the premalignant condition of the human disease and suggests new therapeutic strategies. Dis Model Mech 9, 975-984. ∞ Martinez-Corral I, Stanczuk L, Frye M, Ul- vmar MH, Diéguez-Hurtado R, Olmeda D, Makinen T, Ortega S ( 2016 ). Vegfr3-CreER ( T2 ) mouse, a new genetic tool for target- ing the lymphatic system. Angiogenesis 19, 433-445. ∞ Gardenier JC, Hespe GE, Kataru RP, Savetsky IL, Torrisi JS, Nores GD, Dayan JJ, Chang D, Zampell J, Martínez-Corral I, Ortega S, Mehrara BJ ( 2016 ). Diphtheria toxin-mediated ablation of lymphatic endothelial cells results in progressive lymphedema. JCI Insight 1, e84095. ANNUAL REPORT 2016 144 DIRECTION OF INNOVATION MONOCLONAL ANTIBODIES CORE UNIT OVERVIEW The development of monoclonal antibody (mAb) technology has led to the generation of large panels of highly specific reagents that have had a tremendous impact on basic and applied research over the last four decades. MAbs have become indispensable tools for many of the laboratory techniques that are used to answer essential questions in biomedical research. Their outstanding specificity makes them excellent tools for enabling researchers to better understand biological processes ; particularly in the investigation of new approaches for the diagnosis, prevention and treatment of cancer. The Monoclonal Antibodies Unit provides CNIO Research Groups with an à la carte generation of mAbs. We are highly specialised in the production of mouse and rat mAbs. The Unit also offers mAb production in gene-inactivated mice, mAb characterisation and validation, medium-scale mAb production, and a service of Mycoplasma testing for the cell culture facility. Giovanna Roncador Core Unit Head Technicians Álvaro García ( PEJ )*, Sherezade Jiménez, Lorena Maestre ( TS )**, Ana I. Reyes *Plan de Empleo Joven ( Youth Employment Plan ) **Titulado Superior ( Advanced Degree ) “ The Unit produces novel and high quality mAbs for use in basic research in order to gain new insights into the human cancer development process. We are also highly specialised in mAb characterisation, thereby providing CNIO researchers with reliable and well-validated reagents that give an added value to their research projects.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 145 BIOTECHNOLOGY PROGRAMME | MONOCLONAL ANTIBODIES CORE UNIT RESEARCH HIGHLIGHTS During the last 16 years, the Monoclonal Antibodies Unit has generated a large number of mAbs ( against more than 130 different antigens ), mostly targeting molecules for which mAbs are not commercially available. Many of those mAbs have been licensed to external companies, generating in turn royalties that represent an important source of revenue for the CNIO. Each year we prepare and update a detailed CNIO mAbs catalogue, which contains the datasheets of more than 78 thoroughly validated high-quality mAbs ( accessible at http :// www. cnio. es/ing/servicios/anticuerpos/default.aspx ). This year, in collaboration with the Custom Antibodies Service ( CAbS ) of the Institute for Advanced Chemistry of Catalonia of the Spanish Council for Scientific Research ( IQAC-CSIC ), we have successfully generated several mAbs against small molecules, compounds with low molecular weight such as vitamins, chemicals, hormones, etc., thus expanding our portfolio of reagents in this new field. We also established collaborations with several big pharmaceutical companies ( e.g. Merck, Lilly ) for the production of mAbs against molecules, of their interest, involved in cancer development. EuroMAbNet and its commitment with Ab validation In 2008, in collaboration with Oxford University, we founded EuroMAbNet ( www.euromabnet.com ), a non-profit organisation that includes internationally distinguished multidisciplinary academic laboratories specialised in antibody technologies. Their wealth of expertise ranges from the identification of new targets to the production of fully validated Abs and their use as research tools, clinically relevant diagnostic/prognostic reagents, and novel therapeutics. The use of poorly characterised antibodies is of major concern to the scientific community, resulting in wasted time and valuable research funds, as well as in the publication and perpetuation of erroneous research results, which ultimately compromise the advancement of science. To address this problem, EuroMAbNet has published a position paper ( Roncador et al., 2016 ) and some easy to follow guidelines ( http ://www.euromabnet.com/ guidelines ) that provide a set of criteria and recommendations to help researchers select the most effective mAbs from those available in the market, and provide the strategic guidance needed to perform antibody validation. EuroMAbNet also has a strong commitment to improving the education and training of junior scientists in Ab validation. With that in mind, we have started organising annual Antibody Validation Workshops ( www.euromabnet.com ) to provide practical guidelines about the principles underlying antibody validation, including the verification of Ab specificity, selectivity, sensitivity and reproducibility. These workshops outline the problems generated by the use of poorly validated reagents and educate researchers to minimise the purchase of ineffective Abs. s ∞ PUBLICATIONS ∞ Pérez-Guijarro E, Karras P, Cifdaloz M, Martínez-Herranz R, Cañón E, Graña O, Horcajada-Reales C, Alonso-Curbelo D, Calvo TG, Gómez-López G, Bellora N, Riveiro-Falkenbach E, Ortiz-Romero PL, Rodríguez-Peralto JL, Maestre L, Ronca- dor G, de Agustín Asensio JC, Goding CR, Eyras E, Megías D, Méndez R, Soengas MS ( 2016 ). Lineage-specific roles of the cyto- plasmic polyadenylation factor CPEB4 in the regulation of melanoma drivers. Nat Commun 7, 13418. ∞ Wong KK, Gascoyne DM, Soilleux EJ, Lyne L, Spearman H, Roncador G, Ped- ersen LM, Møller MB, Green TM, Ban- ham AH ( 2016 ). FOXP2-positive diffuse large B-cell lymphomas exhibit a poor response to R-CHOP therapy and dis- tinct biological signatures. Oncotarget 7, 52940-52956. ∞ Roncador G, Engel P, Maestre L, Anderson AP, Cordell JL, Cragg MS, Šerbec VC, Jones M, Lisnic VJ, Kremer L, Li D, Koch-Nolte F, Pascual N, Rodríguez-Barbosa JI, To- rensma R, Turley H, Pulford K, Banham AH ( 2016 ). The European antibody network’s practical guide to finding and validating suitable antibodies for research. MABS 8, 27-36. Figure Antibody validation cloud. ANNUAL REPORT 2016 146 DIRECTION OF INNOVATION MOLECULAR IMAGING CORE UNIT OVERVIEW Molecular imaging involves specialised instrumentation, used alone or in combination with targeted imaging agents, to visualise tissue characteristics and/or biochemical markers. The data generated from molecular imaging studies can be used to help understand biological phenomena, identify regions of pathology, and provide insight regarding the mechanisms of disease. At the Molecular Imaging Unit, we offer state-of-the-art techniques such as Positron Emission Tomography ( PET ), Computed Tomography ( CT ), Ultrasounds ( US ) and Densitometry ( DeXa ). “ Molecular Imaging, especially PET, goes beyond the role of tumour detection and has also taken on the role of tumour characterisation.” Francisca Mulero Core Unit Head Technicians Elena Andrés ( until July ), Guillermo Garaulet ( since April ) ( TS )*, Lucía García ( September- December ), Silvia Leal, Cristina Penalba ( PEJ-L )**, SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 147 BIOTECHNOLOGY PROGRAMME | MOLECULAR IMAGING CORE UNIT RESEARCH HIGHLIGHTS The main objectives of the Unit are to provide CNIO researches with state-of-the-art molecular imaging equipment and human resources in order to : guarantee the highest quality studies, develop and update protocols and techniques to optimise visualisation of tumours in both preclinical and clinical fields, as well as assess and advise researchers on the best-suited imaging modality for their research projects. With the Immuno-PET strategy, the high specificity of the antibody is coupled with the high sensitivity of PET imaging to obtain a strong, non-invasive, tool for glioblastoma ( GBM ) and pancreatic carcinoma diagnosis and follow-up. In 2016, we published the results of our collaboration with the Seve-Ballesteros Foundation Brain Tumour Group and the Crystallography and Protein Engineering Unit at the CNIO. We reported the development of a new tracer (  89Z-LEM2/15 ) for the efficient detection of MT1-MMP in preclinical GBM models. We have also provided imaging support in clinical trials conducted under CNIO’s Clinical Research Programme. With the Breast Cancer Clinical Research Unit, we published the  18F-FMISO-PET imaging results from a clinical trial aimed at selecting patients who will benefit from treatment with angiomodulators knowing the degree of tumour hypoxia by using this PET biomarker ( FIGURE ). Furthermore, we continued our active participation in the international consortium focused on imaging, ‘ M+Visión ’ led by the Massachusetts Institute of Technology ( MIT ). s Elka Jesarela San Martín, Gloria Visdominé *Titulado Superior ( Advanced Degree ) **Plan de Empleo Joven-Licenciado ( Youth Employment Plan-Graduate ) ∞ PUBLICATIONS ∞ Quintela-Fandino M, Lluch A, Manso LM, Calvo I, Cortes J, García-Saenz JA, Gil JM, Martinez-Jañez N, González-Martín A, Adrover E, De Andres R, Viñas G, Llom- bart Cussac A, Alba E, Guerra J, Bermejo B, Zamora E, Moreno-Anton F, Pernas-Simon S, Carrato A, Lopez A, Escudero MJ, Campo R, Carrasco EM, Palacios J, Mulero F and Colomer R ( 2016 ). 18F-fluoromisonidazole PET and activity of neoadjuvant nintedanib in early HER2-negative breast cancer : a window-of-opportunity randomized trial. Clin Cancer Res. PMID : 27587436. ∞ Navarro P, Bueno MJ, Zagorac I, Mondejar T, Sanchez J, Mourón S, Muñoz J, Gómez- López G, Jimenez-Renard V, Mulero F, Chandel NS, Quintela-Fandino M ( 2016 ). Targeting tumor mitochondrial metabo- lism overcomes resistance to antiangio- genics. Cell Rep 15, 2705-2718. ∞ Portal-Núñez S, Ardura JA, Lozano D, Bolívar OH, López-Herradón A, Gutiér- rez-Rojas I, Proctor A, van der Eerden B, Schreuders-Koedam M, van Leeuwen J, Alcaraz MJ, Mulero F, de la Fuente M, Esbrit P ( 2016 ). Adverse Effects of Diabetes Mellitus on the Skeleton of Aging Mice. J Gerontol A-Biol 71, 290-299. ∞ Hernández-Agudo E, Mondejar T, So- to-Montenegro ML, Megías D, Mouron S, Sanchez J, Hidalgo M, Lopez-Casas PP, Mulero F, Desco M, Quintela-Fandino M. ( 2016 ). Monitoring vascular normalization induced by antiangiogenic treatment with ( 18 )F-fluoromisonidazole-PET. Mol Oncol 10, 1704-1718. ∞ Ardura JA, Portal-Núñez S, Lozano D, Gutiérrez-Rojas I, Sánchez-Salcedo S, López-Herradón A, Mulero F, Villanue- va-Peñacarrillo ML, Vallet-Regí M, Esbrit P ( 2016 ). Local delivery of parathyroid hormone-related protein-derived pep- tides coated onto a hydroxyapatite-based implant enhances bone regeneration in old and diabetic rats. J Biomed Mater Res A104, 2060-2070. ∞ de Lucas AG, Schuhmacher AJ, Oteo M, Romero E, Cámara JA, de Martino A, Arroyo AG, Morcillo MÁ, Squatrito M, Martinez-Torrecuadrada JL, Mulero F ( 2016 ). Targeting MT1-MMP as an Im- munoPET-Based strategy for imaging gliomas. PLoS One 11, e0158634. ∞ Muñoz-Mediavilla C, Cámara JA, Salazar S, Segui B, Sanguino D, Mulero F, de la Cueva E, Blanco I ( 2016 ). Evaluation of the foetal time to death in mice after application of direct and indirect euthanasia methods. Lab Animal 50, 100-107. ∞ AWARDS AND RECOGNITION ∞ Scientific Advisory Board Chair and Facul- ty, the Madrid-MIT M+Visión Consortium, Spain. Figure PET-CT imaging with radiolabelled  18F-MISO in patients with breast carcinoma. MISO uptake before treatment ( upper panel ) and after treatment ( lower panel ). We observe the reduction in uptake intensity and the change in the shape of the hypoxic volume after treatment. ANNUAL REPORT 2016 148 DIRECTION OF INNOVATION FLOW CYTOMETRY CORE UNIT OVERVIEW Flow cytometry is a very useful tool in the oncology field. It enables multiparametric analysis for the identification, quantification and isolation of defined subpopulations of cells, based on the levels of expression of fluorescent markers and their relation to each other. Our aim is to provide CNIO Groups with the necessary technical and scientific advice regarding the use of flow cytometric technologies, collaborating with them for the design, acquisition, data analysis and interpretation. With our 4 analysers and 3 high-speed cell sorters, with different configurations of lasers and detectors, we can cater to all our users ’ needs. We also have an automated magnetic bead separation system ( AutoMACS ) and 2 automated cell counters. Analysers are available to users upon appropriate training and cell sorters are operated by the Unit staff. Our sorters can separate up to 4- or 6- defined populations at a time, as well as allow for single cell cloning. We can accept human samples to sort under Biosafety regulations. Lola Martinez Core Unit Head Technicians Ultan P. Cronin ( TS )*, Elena Garrido ( TS )*, Tania López ( PEJ-L )**, Miguel Ángel Sánchez ( TS )* *Titulado Superior ( Advanced Degree ) **Plan de Empleo Joven-Licenciado ( Youth Employment Plan-Graduate ) “ In vivo LacZ detection has always been a challenge. We have optimised a protocol for the identification and isolation of LacZ expressing cells from haematopoietic and lung tissues.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 149 BIOTECHNOLOGY PROGRAMME | FLOw CYTOMETRY CORE UNIT RESEARCH HIGHLIGHTS We provide state-of-the-art equipment and software packages in flow cytometry and collaborate with CNIO investigators in setting up and optimising flow cytometry techniques of their interest. Some of the applications that have been developed and validated by our Unit include : ɗ Cell proliferation studies ( CFSE, Cell Trace Violet, BrdU or EdU, DNA content, etc.). ɗ Apoptosis studies ( Annexin V, Mitochondrial Membrane Potencial, Caspase 3, etc.). ɗ Multicolour Immunophenotyping panels ( B and T cell development, Tregs, Inflammation, etc.). ɗ Functional Assays ( side population detection, Ca 2+ flux, intracellular pH, etc.). ɗ Cytometric Bead Arrays to measure several cytokines from cell extracts and plasma. ɗ Microvesicle detection. We have further developed our multicolour panels for the characterisation of the immune response by incorporating the new generation of Brilliant UV dyes from samples such as haematopoietic tissues, pancreas, skin, liver, and lung. Modifications in our analytical and cell sorters have also been applied to allow for this. Moreover, these panels could still be combined with the detection of proliferation and cell death. In terms of our cell sorting capabilities we included, at the end of the year, a MoFlo ASTRIOS in our portfolio of cell sorters. This cell sorter is equipped with 4 laser lines and 15 fluorescent detectors, which enable the isolation of up to 6 different populations simultaneously. The optical configuration in the ASTRIOS will allow for the use of the new generation of Brilliant UV dyes. s Figure (  A )  Scheme depicting K-RasLSLG 12Vgeo alleles. ( B ) FDG loading protocol scheme. ( C ) Cell viability using DAPI from 6 independent experiments. Error bars represent s.e.m.*, p<0.05, **, p<0.01. ( D ) Representative density plots of the distribution of myeloid and B cells in spleen and lung tissues. ∞ PUBLICATIONS ∞ López-Guadamillas E, Fernández-Marcos PJ, Pantoja C, Muñoz-Martin M, Martínez D, Gómez-López G, Campos-Olivas R, Valverde AM, Serrano M ( 2016 ). p21Cip1 plays a critical role in the physiological adaptation to fasting through activation of PPARa. Sci Rep 6, 34542. ANNUAL REPORT 2016 150 DIRECTION OF INNOVATION CONFOCAL MICROSCOPY CORE UNIT OVERVIEW Optical microscopy has traditionally been an indispensable tool in cell biology studies. In fact, one of the main challenges in oncology research is the study of specific markers, expression patterns or individual cells in the tumour environment. The Confocal Microscopy Unit provides CNIO Research Groups with all the standard methodologies and the latest advances in microscopy. We offer access to state-of-the-art equipment and software packages related to confocal microscopy, including technical and scientific advice and support to the CNIO scientists. The Unit is also actively involved in developing, testing and implementing new microscopy technologies, tools and imaging applications that could be of interest to the Research Groups. Training activities are also an essential component of our mission. “ The Confocal Microscopy Unit is fully committed to disseminating advanced microscopy methodologies that are useful for cancer research in order to benefit society, always with the aim of increasing our understanding of the cell biology and the disorders of cells that cause cancer.” Diego Megías Core Unit Head Technicians Jesús Gómez ( PEJ-L )*, Manuel Pérez ( TS )**, Joaquim Soriano ( TS )** *Plan de Empleo Joven-Licenciado ( Youth Employment Plan-Graduate ) **Titulado Superior ( Advanced Degree ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 151 BIOTECHNOLOGY PROGRAMME | CONFOCAL MICROSCOPY CORE UNIT RESEARCH HIGHLIGHTS The Confocal Microscopy Unit is equipped with 3 laser scanning confocal systems ( Leica SP2 and SP5 ) that incorporate UV and multiphoton excitation, a white light laser and a Hybrid Detector, as well as 2 wide-field systems ( a Deltavision 4D deconvolution station and a Leica DMRI6000 system, equipped with microinjection ). All the microscopes are automated and equipped with incubators for live cell imaging. In addition, the Unit has implemented the use of high-throughput technologies applied to confocal microscopy using 2 different systems : ɗ An Opera ( Perkin Elmer ) High Content Screening ( HCS ) system, which allows running HCS experiments on fixed and live cells in multi-well plates, and enables the monitoring of cell dynamics ( translocation, cell division, etc.) through the use of fluorescence. ɗ A Matrix Screening Application integrated into the SP5 confocal systems, allowing high-throughput feeding of the instrument, not only in multi-well plates, but also in tissue sections. These advances enable us to increase the level of information obtained from a sample as well as carry out the automated screening of cell behaviour under different treatments. During 2016, the Confocal Microscopy Unit contributed to the microscopy field in several aspects. It improved the intelligent screening technique with new algorithms for image acquisition, thereby creating new applications in both confocal and conventional fluorescence microscopy. The use of microfluidics with live-cell assays in perfusion chambers has also experienced a significant increase in performance and demand. In addition, the Unit patented a new device for improving hardware autofocus that will be of great relevance in high-resolution automated image acquisition. Moreover, the Confocal Microscopy Unit continues to dedicate a significant effort towards the development and implantation of High-Content Screening technology at the CNIO ; for example, in 2016, we provided support for the running of screening assays for compounds that could modify mitotic checkpoints, integrity of nucleoli, DNA Damage, BrdU, cell proliferation, etc. Last but not least, in the field of intravital microscopy, we already have several ongoing projects that are focused on metastasis and skin alteration studies. s ∞ PUBLICATIONS ∞ Burén S, Gomes AL, Teijeiro A, Fawal MA, Yilmaz M, Tummala KS, Perez M, Rodri- guez-Justo M, Campos-Olivas R, Megías D, Djouder N ( 2016 ). Regulation of OGT by URI in Response to Glucose Confers c-MYC-Dependent Survival Mechanisms. Cancer Cell 30, 290-307. ∞ Pérez-Guijarro E, Karras P, Cifdaloz M, Martínez-Herranz R, Cañón E, Graña O, Horcajada-Reales C, Alonso-Curbelo D, Calvo TG, Gómez-López G, Bellora N, Riveiro-Falkenbach E, Ortiz-Romero PL, Rodríguez-Peralto JL, Maestre L, Ronca- dor G, de Agustín Asensio JC, Goding CR, Eyras E, Megías D, Méndez R, Soengas MS ( 2016 ). Lineage-specific roles of the cyto- plasmic polyadenylation factor CPEB4 in the regulation of melanoma drivers. Nat Commun 7, 13418. ∞ Andradas C, Blasco-Benito S, Castillo-Llu- va S, Dillenburg-Pilla P, Diez-Alarcia R, Juanes-García A, García-Taboada E, Her- nando-Llorente R, Soriano J, Hamann S, Wenners A, Alkatout I, Klapper W, Rock- en C, Bauer M, Arnold N, Quintanilla M, Megías D, Vicente-Manzanares M, Urigüen L, Gutkind JS, Guzmán M, Pérez-Gómez E, Sánchez C. ( 2016 ). Activation of the orphan receptor GPR55 by lysophos- phatidylinositol promotes metastasis in triple-negative breast cancer. Oncotarget 7, 47565-47575. ∞ Ramírez-Santiago G, Robles-Valero J, Morlino G, Cruz-Adalia A, Pérez-Martínez M, Zaldivar A, Torres-Torresano M, Chichón FJ, Sorrentino A, Pereiro E, Carrascosa JL, Megías D, Sorzano CO, Sánchez-Madrid F, Veiga E8 ( 2016 ). Clathrin regulates lymphocyte migration by driving actin accumulation at the cellular leading edge. Eur J Immunol 46, 2376-2387. ∞ PATENT ∞ Postigo P.A and Megías Vázquez D. ( 2016 ). Uso de un material para la fabricación de un cubreobjetos, un portamuestras o un recipiente de cultivo celular. Spanish Patent Application ES201631277. Figure Whole-mount of ear hair follicles. A global double- fluorescent reporter mouse ( GFP/Tomato ) was used for lineage tracing of epidermal cells. ANNUAL REPORT 2016 152 DIRECTION OF INNOVATION PROTEOMICS CORE UNIT OVERVIEW Proteins catalyse and control almost all cellular processes in a living cell. The levels of protein abundance, together with their modification states and interactions, adapt dynamically to external or internal ( genetic ) stimuli and thus define the cell’s functional state and determine its phenotype. Recent developments in sample preparation, liquid chromatography, mass spectrometry and data analysis have enabled researchers to investigate diverse proteomic facets in a systematic high- throughput manner, currently comparable to next-generation sequencing platforms. As a result, proteomics is positioned as one of the most powerful technologies to study, at the protein level, complex cellular processes. This vast amount of data is providing new insights into the molecular mechanisms underlying diverse human pathologies such as cancer. “ Mass spectrometry-based technologies enable probing the composition, structure, function and regulation of the proteome, providing new insights into the underlying mechanisms of cancer.” Javier Muñoz Core Unit Head Graduate Student Ana Martínez SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 153 BIOTECHNOLOGY PROGRAMME | PROTEOMICS CORE UNIT RESEARCH HIGHLIGHTS Throughout 2016, the Unit continued its mission of implementing and optimising quantitative proteomic strategies. More specifically, we have introduced a new fractionation method using high pH reverse phase micro columns, which minimises sample loss and thus is highly suitable for low amounts of material. We used this approach to post-fractionate samples enriched in phosphopeptides, substantially increasing the number of identifications. This workflow was used to determine phosphorylation dynamics upon activation of WT and kinase- mutant platelets ( in collaboration with the Cell Division and Cancer Group ), as well as to identify potential substrates of CDK8 involved in the establishment of ground state pluripotency ( in collaboration with the CNIO Tumour Suppression Group ). More recently, in collaboration with the Metabolism and Cell Signalling Group at the CNIO, we also used phosphoproteomics to better understand the molecular mechanism of the mTOR pathway. Together with the CNIO Genomic Instability Group, we are using a recent approach, named Thermal Proteome Profiling ( Savitski et al., see FIGURE ), to identify protein targets of certain inhibitors ( e.g. target deconvolution ). We have also performed several AP-MS/MS experiments for different proteins ( STAG1, STAG2, PDS5A, PDS5B ) belonging to the cohesion complex ( with CNIO’s Chromosome Dynamics Group ). Likewise, we have identified a large protein network ( more than 300 proteins ) that interacts with the RNA pol II complex ( in collaboration with the Tumour Suppression Group ). Over the last few years, the analysis of the protein content of exosomes has received great interest in the context of metastasis and the pre-metastatic niche. Along this line, we are conducting several proteomic analyses of exosomes from different origins in collaboration with CNIO’s Microenvironment and Metastasis Group, the Gastrointestinal Cancer Clinical Research Unit and the Melanoma Group. s Technicians Fernando García ( TS )*, Nuria Ibarz ( TS )*, Ailyn Martínez ( since April ), M. Isabel Ruppen ( TS )*, Pilar Ximénez de Embún ( TS )*, Eduardo Zarzuela ( TS )* ( PEJ-L )** *Titulado Superior ( Advanced Degree ) **Plan de Empleo Joven-Licenciado ( Youth Employment Plan-Graduate ) Student in Practice Julia Beltran ( since October ) ∞ PUBLICATIONS ∞ Lecona E, Rodriguez-Acebes S, Specks J, Lopez-Contreras AJ, Ruppen I, Murga M, Muñoz J, Mendez J, Fernandez-Capetillo O ( 2016 ). USP7 is a SUMO deubiquitinase essential for DNA replication. Nat Struct Mol Biol 23, 270-277. ∞ Navarro P, Bueno MJ, Zagorac I, Mondejar T, Sanchez J, Mourón S, Muñoz J, Gómez- López G, Jimenez-Renard V, Mulero F, Chandel NS, Quintela-Fandino M ( 2016 ). Targeting Tumor Mitochondrial Metabo- lism Overcomes Resistance to Antiangi- ogenics. Cell Rep 15, 2705-2718. ∞ Martinez-Val A, Garcia F, Ximénez-Embún P, Ibarz N, Zarzuela E, Ruppen I, Moham- med S, Munoz J ( 2016 ). On the Statistical Significance of Compressed Ratios in Iso- baric Labeling : A Cross-Platform Com- parison. J Proteome Res 15, 3029-3038. Figure Thermal proteome profiling ( TPP ). Cells are treated either with the compound of interest or the vehicle. Cells are subjected to increasing temperatures, and denatured proteins are discarded by centrifugation. Supernatants are analysed by LC-MS/MS enabling the reconstruction of the melting curves for all identified proteins. Proteins showing a difference in their melting point between drug and vehicle might be potential targets. ANNUAL REPORT 2016 154 DIRECTION OF INNOVATION HISTOPATHOLOGY CORE UNIT “ The Histopathology Core Unit participates in several External Quality Assessment Schemes, such as NordiQC and UKNEQAS, which independently evaluate the quality of the techniques performed at the Unit. In 2016, several protocols developed by the Unit were incorporated into the Best Methods section by the UKNEQAS.” Alba De Martino Core Unit Head Technicians Nuria Cabrera, María Gómez, Patricia González, Gabino Hernández, Verónica Neva ( PEJ )*, Paula Tejedor ( since September ), Irene Roda ( since October ), Maria Udriste, Zaira Vega *Plan de Empleo Joven ( Youth Employment Plan ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 155 BIOTECHNOLOGY PROGRAMME | HISTOPATHOLOGY CORE UNIT OVERVIEW Pathology is devoted to the study of the structural, biochemical and functional changes in cells, tissues and organs that underlie disease. By using molecular, immunological and morphological techniques, pathology serves as the bridge between the basic sciences and clinical medicine. The Histopathology Core Unit offers knowledge and expertise through a full range of services encompassing paraffin embedding and cutting, as well as the construction of tissue microarrays ( TMAs ). We also provide our users with histochemical stains upon request, research and diagnostic immunohistochemistry ( IHC ) testing, antibody validation, and in situ hybridisation ( ISH ) ( ALU sequences for mouse xenograft characterisation ). Furthermore, the Unit offers other services, such as laser-capture microdissection ; slide digitalisation for brightfield, polarisation light and fluorescence ; image analysis ; and quantification. The Unit collaborates with researchers at any stage of their career in the histological characterisation of phenotypically relevant animal models of disease, thus providing them with the Pathology expertise required for the success of their research projects. RESEARCH HIGHLIGHTS In 2016, the Unit beat previous records in the Histopathology database with more than 15,000 new entries. This corresponded to about 40,000 requests processed with approximately 37,000 paraffin-embedded blocks ; 40,000 histochemical techniques performed ; 13,000 routine immunohistochemistry techniques performed ( not counting optimisation tests ) ; 7,000 scanning requests for histological slide scanning and image analysis ; and 70 requests for laser microdissection. All the developed techniques follow a standardised validation process. In 2016, the Unit added several new antibodies to its portfolio, which includes more than 3,000 tested and 1,000 currently available antibodies that have been optimised for both human and mouse tissue samples. The antibody validation process follows rigorous testing in order to achieve the best possible results and to demonstrate reproducibility between assay runs and between batches. This represents a highly valuable resource for CNIO researchers as well as for the external clinical and research community. In respect of the importance that our researchers place on quality and reproducibility, our Unit participates in several External Quality Assessment Schemes, such as NordiQC and UK NEQAS, which evaluate the quality of the staining techniques performed at the Unit and in which more than 800 laboratories participate worldwide. In 2016, our Unit scored very high in the evaluated techniques, and several protocols developed by the Unit were incorporated into the ‘ Best Methods section ’ of the UKNEQAS Cellular Pathology Technique website ( PAS staining and Haematoxylin-Eosin, among others ). s Figure Techniques developed at the Histopathology Core Unit and routinely used by CNIO researchers. Sirius Red, Lung. Brightfield ( top left ) and Sirius Red, lung. Polarised light ( middle left ). Immunofluorescence, skin ( bottom left ). Luxol Fast Blue staining, decalcified spinal cord ( top right ). Improved ALU II in situ hybridisation for mouse xenograft detection ( bottom right ). ∞ PUBLICATIONS ∞ Mosteiro L, Pantoja C, Alcazar N, Marión RM, Chondronasiou D, Rovira M, Fernan- dez-Marcos PJ, Muñoz-Martin M, Blan- co-Aparicio C, Pastor J, Gómez-López G, De Martino A, Blasco MA, Abad M, Serrano M ( 2016 ). Tissue damage and senescence provide critical signals for cellular reprogramming in vivo. Science 354, 6315. pii : aaf4445. ∞ Hernández-Porras I, Schuhmacher AJ, Garcia-Medina R, Jiménez B, Cañame- ro M, de Martino A, Guerra C ( 2016 ). K-Ras( V14I ) -induced Noonan syndrome predisposes to tumour development in mice. J Pathol 239, 206-217. ∞ de Lucas AG, Schuhmacher AJ, Oteo M, Romero E, Cámara JA, de Martino A, Arroyo AG, Morcillo MÁ, Squatrito M, Martinez-Torrecuadrada JL, Mulero F ( 2016 ). Targeting MT1-MMP as an Im- munoPET-Based strategy for imaging gliomas. PLoS One 11, e0158634. ANNUAL REPORT 2016 156 DIRECTION OF INNOVATION ANIMAL FACILITY “ The accreditation of our animal research programme by the AAALAC reflects CNIO’s compromise and high level of excellence with respect to the care and use of animal models, which are essential in cancer research.” Isabel Blanco Core Unit Head Management Vivotecnia Management & Services SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 157 BIOTECHNOLOGY PROGRAMME | ANIMAL FACILITY The CNIO has a state-of-the-art Animal Facility, managed by Vivotecnia Management & Services. The Animal Facility’s primary responsibility is the supply, husbandry and quality control of laboratory animals used by the Research Programmes in their experimental protocols. The strict compliance to national, EU and international recommendations regarding the use and care of animals in research is of paramount importance to the CNIO. The high standards achieved by the CNIO with regards to the use and care of animals for experimentation have been recognised by the Association for Assessment and Accreditation of Laboratory Animal Care International ( AAALAC ); they granted us full accreditation in October 2016. AAALAC International is a private non-profit organisation that promotes the humane treatment of animals in science through voluntary accreditation and assessment programmes. More than 950 companies, universities, hospitals, government agencies and other research institutions across 41 countries have earned AAALAC accreditation, which is considered one of the top international recognitions in this field. The CNIO Animal Facility was established to assist researchers in the development and analysis of in vivo models. We are currently collaborating with as many as 25 CNIO Research Groups, Sections and Units from different Research Programmes. Our Animal Facility has the capacity to house 19,000 type IIL cages. Our mouse lines are maintained and bred in the Facility’s barrier area, which assures Specific Pathogen Free ( SPF ) health status through a comprehensive health surveillance programme. Microbiological and environmental parameters in the animal areas are constantly monitored. All mouse strains housed in the barrier are either generated within the barrier or introduced by rederivation. We also have an additional area with a capacity for 1,800 type II cages dedicated for the use of non-replicative strains of adenovirus, lentivirus and retrovirus, as well as for xenograft models. In this area, mice are housed in ventilated racks with integration of Individually Ventilated Caging ( IVC ) units in the building ventilation systems. Mice are always manipulated in Type II biosafety cabins. Daily operations and husbandry procedures are highly automated in order to safe-guard our personnel from any associated risks ; robotic devices perform the potentially hazardous tasks such as the processing of dirty bedding, the washing/filling of cages and bottles, etc. These automated systems maximize the productivity and ensure the quality standards in our washing and sterilising areas. All records concerning breeding protocols and animal inventory are computerised and stored in a web-based application accessible via the CNIO intranet. The Animal Facility has harboured more than 2,500 genetically modified mouse lines ; currently, there are more than 700 genetically modified lines and more than 45,000 live mice. The Facility also provides access to more than 80 tool strains, including constitutive and inducible Cre strains, Flp strains, reporter strains, Tet transactivator strains and others. The Animal Facility offers the possibility of running a broad number of experimental procedures in the premises, including the use of gamma irradiation, UV light and volatile carcinogenic agents, as well as surgical procedures, some behavioural studies, a non-invasive blood pressure system, and a lab animal monitoring system ( Oxylet ) that allows measuring a number of physiological parameters for metabolic profiling and phenotyping of mouse models. Additionally, the monitoring of the mouse models through non- invasive imaging technologies is provided by the Molecular Imaging Core Unit, which has integrated all its image acquisition instruments within the Animal Facility. Likewise, the work of the Transgenic Mice Unit is performed in a laboratory inside the SPF barrier. Finally, the necropsy laboratory is equipped with instruments for the haematological and biochemical analysis of blood and urine, which complement the pathology and clinical diagnostics. All the work carried out by the Animal Facility complies with both national and EU legislation –Spanish Royal Decree RD53/2013 and EU Directive 2010/63/UE– for the protection of animals used for research experimentation and other scientific purposes. Experimental procedures and projects are reviewed and evaluated by the Research Ethics and Animal Welfare Committee of the Instituto de Salud Carlos III, as well as by the Institutional Animal Care and Use Committee ( IACUC ). The Orden ECC/566/2015 stipulates that all animal procedures are to be carried out by qualified people in the possession of the corresponding accreditation as issued by the competent authority. Currently, the Animal Facility Service is performed by more than 35 qualified persons between care workers, technicians, supervisors and veterinarians involved in the breeding and care of animals, ensuring the welfare of the animals. The Animal Facility offers CNIO’s new staff a short course focused on the training of personnel performing work with laboratory animals ; this is complementary to the online courses that are a requisite to gain access to the facility. In line with our commitment to maintain the highest possible standards in regards to animal research issues, the CNIO has joined the Agreement on Openness on Animal Research promoted by the Federation of Scientific Societies in Spain ( COSCE ), in collaboration with the European Animal Research Association ( EARA ), which was launched on September 2016. An institutional statement on the use of animals in research can be consulted in the CNIO website. s ∞ PUBLICATION ∞ Muñoz-Mediavilla C, Cámara JA, Salazar S, Segui B, Sanguino D, Mulero F, de la Cueva E, Blanco I ( 2016 ). Evaluation of the foetal time to death in mice after application of direct and indirect euthanasia methods. Lab Animal 50, 100-107. ANNUAL REPORT 2016 158 DIRECTION OF INNOVATION EXPERIMENTAL THERAPEUTICS PROGRAMME JOAQUÍN PASTOR Programme Director “ With a proven track record in the development of advanced compounds for cancer therapy, The Experimental Therapeutics Programme (ETP) is now aligning its capabilities with CNIO’s growing research on phenotypic drug discovery. Consequently, we are contributing with state-of-the-art approaches to Target Deconvolution activities.” SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 159 EXPERIMENTAL THERAPEUTICS PROGRAMME The current ETP-CNIO pipeline encompasses targeted and phenotypic projects. The following highlights summarise some of our achievement during 2016. Our most advanced targeted project is dedicated to CDK8 inhibitors. We have selected our first lead, ETP-27, which has yielded positive results in PK-PD studies and has shown early signs of efficacy in a MOLM13 xenograft model after oral administration. Currently, we have embarked on the fine optimisation of the in vivo exposure within this series. Additional targeted projects, focused on Mastl and Haspin inhibitors, are undertaken in collaboration with Marcos Malumbres ( CNIO Cell Division and Cancer Group ). The Mastl project started with a cell-based screening of several ETP-libraries, where a few ‘ high-micromolar ’ hits emerged as potential Mastl inhibitors. However, a direct target engagement experiment to unequivocally identify Mastl as their molecular target was not possible due to the unavailability of isolated active Mastl protein. ETP’s Biology Team has been able to set up highly efficient conditions for the purification, isolation and production of ‘ full length active Mastl kinase ’. This important achievement has enabled, for the first time, the biochemical profiling of the identified hits and an additional targeted biochemical screening of ETP-libraries. As a result of these activities, we have identified several families of compounds as biochemical Mastl inhibitors, including ETP-750 with an IC50 of around 300 nM. These results shall pave the way for the discovery and development of advanced Mastl inhibitors in the near future. It is worth mentioning that the production of the protein was carried out by the CNIO Crystallography and Protein Engineering Unit. Using the same cell-based assay, Malumbres ’ Group identified compounds that efficiently produce ‘ mitotic cell death ’, a new avenue for cancer therapy. The knowledge of the biochemical profile of these hits obtained by ETP has contributed to the identification of Haspin kinase as the target responsible for the observed phenotype. Currently, ETP is working on the discovery of Haspin inhibitors where we have already identified highly potent compounds in the low-nanomolar range. We are also collaborating with P. Carmeliet ( VIB-KU Leuven, Belgium ) for the discovery of novel inhibitors of a particular enzyme in the field of vascular normalisation. We have carried out a screening campaign and we are now working on hit generation activities to identify catalytic inhibitors of this enzyme and their associated intellectual property. Phenotypic screenings have proven to be advantageous for the discovery of innovative molecular targets and modulators, as well as to establish their link with disease. Nevertheless, the molecular target responsible for a desired phenotype needs to be identified and this deconvolution phase implies an extra step of complexity in the process. As mentioned above, we are working on several phenotypic projects : ETP collaborates with Manuel Serrano ( CNIO Tumour Suppression Group ) in a project dedicated to the discovery of novel targets and modulators against Cancer Stem Cells ( CSCs ). We have been focusing on target deconvolution activities around previously identified hits that have the ability to selectively kill CSCs and to inhibit the tumour-initiating capacity of pancreatic CSCs. Our Medicinal Chemistry Team has designed and synthesised chemical probes around those molecules by using ‘ minimalist linkers ’. These chemical fragments bear a photoactivatable group to achieve crosslinking of the modified hits with targeted proteins, and a special chemical group to perform ‘ click chemistry ’ that is useful to attach the ‘ cross- linked complex ’ to a reporter tag for imaging and/or pull-down experiments. The treatment of cells and cell lysates with these molecules has enabled the identification of a target candidate, which is currently under additional validation studies. ETP is collaborating with Maria A. Blasco ( CNIO Telomeres and Telomerase Group ) in the discovery of TRF1 inhibitors, a project that also requires a ‘ target deconvolution ’ phase. During this year, we focused our attention on a series of TRF1 inhibitors with an unknown and perhaps innovative mechanism of TRF1 modulation. We have profiled the main hit, ETP-946, against large panels of enzymes and receptors representing more than 600 targets. Among them, we have retrieved 4 potential candidates, which are currently undergoing validation studies. The chemical modification of ETP-946 with ‘ minimalist linkers ’ has yielded potential affinity probes that are currently under evaluation. Last but not least, ETP has set up a ‘ thermal stability assay ’ to study the stabilisation of hTRF1 overexpressed in HEK293 cells in the presence of ETP-946 and analogues. These experiments will inform us about direct interactions of those compounds with TRF1. Finally, I’d like to mention that other ‘ phenotypic exploratory projects ’ are also currently at the screening phase ; these are undertaken in collaboration with the CNIO Researchers Manuel Valiente, Óscar Fernández-Capetillo and Massimo Squatrito. ANNUAL REPORT 2016 160 DIRECTION OF INNOVATION MEDICINAL CHEMISTRY SECTION Sonia Martínez Section Head Staff Scientists Ana Belén García, Cristina Gómez, Esther González, Ana Isabel Hernández, María del Rosario Rico, Sonsoles Rodríguez, Carmen Varela SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 161 EXPERIMENTAL THERAPEUTICS PROGRAMME | MEDICINAL CHEMISTRY SECTION OVERVIEW The Medicinal Chemistry Section is part of the Experimental Therapeutics Interdisciplinary Programme that is dedicated to early Drug Discovery activities in the oncology field. Our mission is to discover and to develop new anticancer agents based on novel hypotheses and targets generated by CNIO’s Basic Research Groups, and to do so in close collaboration with these groups. Medicinal chemists are responsible for the design, the preparation and the optimisation of compounds for biological evaluation, as well as for the generation of Structure Activity Relationships ( SARs ) and the development of novel lead compounds with in vivo activity in appropriate animal models. Recently, and due to the increasing number of projects related to the cell phenotype, our Programme is entering the chemical biology field in order to contribute to the deciphering of the precise protein target or mechanisms of action responsible for the observed phenotypes. In this regard, we chemically modify the active molecules into affinity probes − compounds with appropriate linkers that form either reversible or irreversible complexes with their targets inside living cells and/or cell lysates − that enable us to directly identify the target/s by extracting them via pull-down experiments, followed by mass spectrometry resolution and cellular localisation experiments using imaging techniques. “ We have successfully designed and synthesised affinity-based chemical probes used in imaging and pull-down/LC-MS/MS analysis experiments in order to discover novel targets/ inhibitors of CSC proliferation, in collaboration with the CNIO Tumour Suppression Group.” RESEARCH HIGHLIGHTS During 2016, our Section was involved in several projects at different phases of the drug discovery process, among them : Cyclin-dependent protein kinase 8 inhibitors ( CDK8i ) project In this funded project ( grant no. SAF2013-44267-R ), we have identified lead compound ETP-27, which has served to demonstrate in vivo Proof of Concept ( PoC ) for CDK8 inhibition in cancer with positive preliminary results. This chemical series was protected by a patent application that reached PCT level in August 2016 and has been exemplified during 2016. Additionally, crystallographic studies with ETP-27 and some analogues from this chemical series have been performed by the CNIO Crystallography and Protein Engineering Unit confirming the expected binding mode of the molecules in the catalytic site. Currently, we are dedicated to the fine optimisation of the lead compound, trying to increase the oral exposure levels vs time. Modifications in the molecule to reduce Clearance or to increase the Volume of Distribution by introducing basicity, for example, are being considered. The final objective of the project is to obtain an advanced product that is ready for preclinical regulatory development and further clinical studies. Graduate Student Francisco J. García Technicians Ivan Arribas ( PEJ-L )*, Carmen Fernández ( PEJ-L )*, Virginia Rivero ( until February ) ( TS )**, Sandra Sanz ( PEJ )*** *Plan de Empleo Joven-Licenciado ( Youth Employment Plan-Graduate ) **Titulado Superior ( Advanced Degree ) ***Plan de Empleo Joven ( Youth Employment Plan ) ANNUAL REPORT 2016 162 DIRECTION OF INNOVATION Microtubule-associated serine/threonine protein kinase-like ( MASTL ) inhibitors A chemical exploration around the hit identified in the biochemical High-Throughput Screening ( HTS ) with active full length human MASTL protein has been set up ; the aim is to define the pharmacophore required for Mastl activity and to increase the activity of current hits, in order to obtain more potent inhibitors that can be used in biological assays as tool compounds. We will then use this information for the design of novel Mastl inhibitors, including Intellectual Property in their structures. HASPIN inhibitors Haspin inhibitors that produce a rapid and efficient mitotic cell death have been identified. We have started a chemical programme in order to explore the current hits and also to generate novel compounds. We are exploring 2 chemical series with haspin inhibitory activity in the low nanomolar range, but also with several main off-targets. Synthesis of analogues has allowed us to learn how selectivity can be achieved without affecting haspin activity. Crystallographic studies of 2 hits from different chemical series have been performed by the CNIO Crystallography and Protein Engineering Unit. Kinase X * inhibitors We finalised the Hit-to-Lead phase in collaboration with VIB ( Belgium ) and have obtained a novel chemical series, in which we identified a potent compound with controlled selectivity and oral bioavailability. The compound has been delivered to VIB to be characterised in different in vivo studies. Kinase Y * inhibitors We are collaborating with VIB for the generation of novel inhibitors of a particular kinase. Several reference compounds and analogues have been synthesised in order to help with the validation studies. After analysis of the hits from an HTS campaign, we have concluded that they were not good enough as starting points for further exploration. Therefore, a hit generation plan has been designed and is currently ongoing. Inhibition of Cancer Stem Cell ( CSC ) proliferation In a collaborative project with the CNIO Tumour Suppression Group, we identified several hits that are able to modulate CSC proliferation, stemness and, at sublethal doses, inhibit the tumour Figure 1 ( A ) Affinity probes ( ETP ) for target identification of inhibitors of CSC proliferation. After SAR we identified a particular basic fragment that is essential for activity. Affinity probes were synthesised ; they included this fragment and a minimalist linker in their design. As a negative control, we synthesised the inactive probe, in which the basicity was removed. ( B ) Overall workflow ( performed by Tim Cash, from the Tumour Suppression Group ). Incubation of the affinity probe in cell culture ( living cells and/or cell lysates ) followed by photoirradiation to capture close proteins. Subsequent click chemistry with Rhodamine-N3 and/ or Avidin Beads allows for imaging and pull-down experiments. *confidential SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 163 EXPERIMENTAL THERAPEUTICS PROGRAMME | MEDICINAL CHEMISTRY SECTION initiating capacity of pancreatic CSCs. In order to decipher the target behind the observed phenotype, we have performed chemotype searches linking the structure with potential targets and profiling in broad panels of enzymes and receptors. As a result of the chemical exploration, we have found that the presence of basicity in these molecules is essential for activity. Based on this discovery, we have been able to successfully synthesise different affinity probes by adding a ‘ minimalist linker ’ to the basic centre, retaining the required cellular activity. These modified molecules, after treatment with cells or cell lysates and photo-irradiation, can covalently capture their binding proteins in a distance- dependent manner. The subsequent click chemistry reaction of the terminal alkyne group of the linker with different reporters ( i.e. rhodamine-N3 or biotin-N3 ) enables, via pull-down experiments, the identification of potential cellular protein targets of the drug, as well as imaging-based determination of their cellular localisation. Because these experiments require the use of appropriate controls, we have also been able to synthesise inactive analogues by removing the basicity of the hit and have performed similar pull down and imaging experiments. The already identified candidate targets are going through a validation process. Telomeric repeat binding factor 1 ( TRF1 ) inhibitors This project is undertaken in collaboration with the CNIO Telomeres and Telomerase Group. After a screening campaign, using a cell-based assay to measure the removal of TRF1 from telomeres, we have identified several hits, among them ETP946. Our main objective during 2016 has been both the deconvolution of the molecular target behind the observed effect using this hit, as well as the chemical exploration to increase SAR knowledge within this chemical series. In the deconvolution studies, we carried out several chemotype searches and an extensive profiling of the compound against a broad range of enzyme and receptor panels. From these studies we have identified 4 candidate targets that are currently under study. Also, we have generated the first affinity probes by the introduction of appropriate linkers in the molecule that will allow for reversible or irreversible interactions with their molecular targets. Currently, we are testing these probes to address their TRF1 modulation in cells. If they show activity they will be very useful for further imaging localisation and pull- down experiments in order to identify the responsible molecular targets for TRF1 modulation, including TRF1 itself. s ∞ PUBLICATIONS ∞ López-Gudamillas E, Muñoz M, Martinez S, Pastor J, Fernandez-Marcos PJ, Serrano, M ( 2016 ). PI3Ka inhibition reduces obesity in mice. Aging 8, 2747-2753. ∞ Aragoneses-Fenoll L, Montes-Casado M, Ojeda G, Acosta YY, Herranz J, Martínez S, Blanco-Aparicio C, Criado G, Pastor J, Dianzani U, Portolés P, Rojo JM ( 2016 ). ETP-46321, a dual p110a/δ class IA phos- phoinositide 3-kinase inhibitor modulates T lymphocyte activation and collagen-in- duced arthritis. Biochem Pharmacol 106, 56-59. ∞ PATENT ∞ Pastor J. et al. ( 2016 ). New Compounds. PCT/GB2016/052641. Figure 2 Experimental Therapeutics Programme Pipeline. ANNUAL REPORT 2016 164 DIRECTION OF INNOVATION BIOLOGY SECTION Carmen BlancoSection Leader Staff ScientistOliver Renner SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 165 EXPERIMENTAL THERAPEUTICS PROGRAMME | BIOLOGY SECTION OVERVIEW In the Experimental Therapeutics Programme, we are working on both targeted and phenotypic-based drug discovery projects. In the targeted projects, the Biology Section is devoted to the biochemical, cellular, and in vitro/in vivo pharmacological characterisation of the compounds synthesised within the Programme. Our aim is to obtain novel anticancer agents with optimised profiles that are able to demonstrate in vivo proof of concept in animal models of cancer. In phenotypic-based projects, we perform target engagement and target deconvolution tasks using label-free techniques and chemical probes, respectively. The cellular thermal shift assay is a label-free technology that enables drug binding studies to target proteins in their relevant cellular contexts. Alternatively, we use engineered chemical probes to deconvolute the possible targets of our molecules. For that purpose, we first have to confirm that they behave similarly to the parent compound in the phenotypic screening. These molecules bear a linker with functionalities that permit their crosslinking with targeted proteins and the attachment of reporter tags for imaging and pull down experiments in cells and cell lysates. Thus, through immunofluorescence assays, we can determine the cellular localisation of the complex of the chemical probe with its targets and, by pull down experiments, using active and inactive chemical probes also in competition with the parent compound followed by mass spectrometry analysis, we can identify candidate targets for the observed phenotype. Furthermore, in exploratory screening projects carried out in collaboration with other CNIO Groups, we provide support by preparing customised compound assay plates from our ETP-libraries, adding the compounds using automated liquid-handling instruments that allow rapid, accurate, and reproducible compound dispensing and assay plate setup. All this instrumentation is integrated in a platform that allows sample tracking and recording. “ Upon production of active human full length MASTL protein, with the support of the CNIO Crystallography and Protein Engineering Unit, we have set up a biochemical assay that has allowed us to perform both target engagement validation with the hits obtained from a MASTL phenotypic screening, and a High- Throughput screening where we have identified a MASTL inhibitor with an IC50 of 300nM among other hits.” Post-Doctoral Fellows Borja Barrera ( until January ), Elena Hernández ( since October ) Technicians Enara Aguirre ( until February ) ( TS )*, M. Isabel Albarrán ( TS )*, Adrián Amezquita ( PEJ )**, Antonio Cebriá ( TS )*, Jennifer García ( PEJ )**, Elena Gómez-Casero ( TS )*, Javier Klett ( TS )*, M. Carmen Rodríguez De Miguel ( TS )* *Titulado Superior ( Advanced Degree ) **Plan de Empleo Joven ( Youth Employment Plan ) ANNUAL REPORT 2016 166 DIRECTION OF INNOVATION RESEARCH HIGHLIGHTS During 2016, our Section was involved in several projects : Cyclin-dependent kinase 8 ( CDK8 ) This is a funded project ( grant no. SAF2013-44267-R ). We have identified ETP-27, a highly selective picomolar CDK8/CDK19 inhibitor with picomolar cellular inhibition of P-STAT1-S727. The main off-target identified is Haspin ; ETP-27 demonstrates a cellular selectivity of > 30 fold for CDK8 vs. Haspin. This compound shows good solubility, permeability, metabolic stability in human and rat microsomes that is moderate in the mouse, and no alerts in terms of CYP-P450 inhibition and hERG binding, as well as important toxicity related parameters. Moreover, the compound is orally bioavailable as observed in pharmacokinetic ( PK ) studies in mice ; in mechanistic PK-PD studies, modulation of biomarker PSTAT1-S727 was observed between 1 and 4 h after oral treatment. ETP-27 has shown promising results in an efficacy study in MOLM13 xenografts, where it demonstrated a 50% tumour growth inhibition after 10 mg/kg twice-a-day treatment over 16 days. Our next steps will focus on the in vivo testing of new compounds resulting from the fine optimisation of ET-27 in order to achieve an even higher in vivo exposure. Microtubule-associated serine/threonine protein kinase-like ( MASTL ) This project is undertaken in collaboration with the CNIO Cell Division and Cancer Group. We have previously reported the Figure From phenotypic- to target-based drug discovery ; the MASTL case project. An HTS screening, together with a phenotypic screening designed to identify compounds that allow exit from mitosis of mitotically arrested cells, was used to identify potential MASTL inhibitors. Five hits were identified. To perform target engagement we produced and purified hMASTL full length protein and set up a biochemical assay. We confirmed 3 hits as MASTL inhibitors with an IC50 between 10 and 50 µM. We performed a screening of 800 compounds with the biochemical targeted assay identifying a MASTL inhibitor with an IC50 of 300nM, which is a good starting point for SAR exploration and pharmacophore development. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 167 EXPERIMENTAL THERAPEUTICS PROGRAMME | BIOLOGY SECTION production of active human full length MASTL protein to run biochemical assays. We used it to validate the hits coming from the phenotypic screening and to perform a biochemical screening of our ETP-640 library. In a single point screening assay, with the cut-off value set at 40% inhibition, we achieved a hit rate of 0.02. Hits were confirmed in a dose response assay and a number of analogues were tested. We have identified a 300nM hit, which is a good starting point for the exploration of Structure-Activity- Relationships ( SAR ) and for pharmacophore development. This information will be used later on for hit generation and subsequent hit-to-lead ( HtL ) exploration of novel inhibitors. Kinase X This project is a collaborative undertaking with VIB ( Belgium ). We have identified a lead compound with nanomolar inhibition activity for the target, good solubility, permeability and high selectivity against a panel of 456 kinases. Moreover, the compound is orally bioavailable, well tolerated in mice and, in a distribution study, was detected in several tissues for up to 8 hours. This lead compound is being used at VIB for in vivo validation studies. Kinase Y In this second project, also in collaboration with VIB, we have characterised at the biochemical level, 30 compounds synthesised in hit generation activities. Telomeric repeat binding factor 1 ( TRF1 ) This project is carried out in collaboration with the CNIO Telomeres and Telomerase Group. A phenotypic assay to measure the association of TRF1 to telomeres has been used to test 72 compounds that include analogues of a second hit, ETP-946, identified in the initial screening ; chemical probes derived from it have been generated. Moreover, we have set up a cellular thermal shift assay with over-expressed human TRF1 to validate if this series of compounds interacts directly with TRF1, and with potential to serve as a platform for identifying compounds that directly interact with TRF1. Cancer stem cells ( CSC ) and gluconeogenesis These projects are carried out in collaboration with the CNIO Tumour Suppression Group. For the CSC project, we collaborated in the deconvolution studies of the identified hits, evaluating their activity in kinase and receptor panels, as well as providing technical support for the assays using chemical probes. For the gluconeogeneis project, we have performed pharmacokinetic profiles of the hits obtained in the screening ; the most promising compound has been tested in vivo by our collaborator. Brain metastasis screening The CNIO Brain Metastasis Group has developed an ex vivo assay to search for drugs that kill human brain metastasis in mice. ETP-Biology has provided support in running the experiments and also to validate the in vitro screening results obtained with the approved or in clinical trial ETP-antitumour library. Two classes of drugs are under further characterisation. Focal adhesion kinase ( FAK ) This project is undertaken in collaboration with the CNIO Cell Signalling and Adhesion Group. We have set up a biochemical assay with purified protein produced by our collaborator, using both the catalytic domain alone and the full length protein. We have tested 80 compounds coming from a virtual screening analysis ; the hit rate obtained was of 0.21 with a cut off value set at 90% inhibition. Five of the compounds with an IC50 below 500nM have been selected for crystallisation studies. Support to other CNIO Groups We have given support to Manuel Serrano’s Group by analysing, with liquid chromatography-tandem mass spectrometry ( LC-MS/ MS ), in tumour and host-mouse plasma samples, the levels of a standard-of-care-drug administered in nanoparticles in order to improve the delivery of chemotherapeutics to their site of action. s ∞ PUBLICATIONS ∞ Mosteiro LL, Pantoja C, Alcazar N, Marión RM, Chondronasiou D, Rovira M, Fernan- dez-Marcos PJ, Muñoz-Martin M, Blan- co-Aparicio C, Pastor J, Gómez-López G, de Martino A, Blasco MA, Abad M, Serrano M ( 2016 ). Tissue damage and senescence provide critical signals for cellular reprogramming in vivo. Science 354, 6315. pii : aaf4445. ∞ Jiménez-García MP, Lucena-Cacace A, Robles-Frías MJ, Narlik-Grassow M, Blan- co-Aparicio C, Carnero A ( 2016 ). The role of PIM1/PIM2 kinases in tumors of the male reproductive system. Sci Rep 6, 38079. ∞ Aragoneses-Fenoll L, Montes-Casado M, Ojeda G, Acosta YY, Herranz J, Martínez S, Blanco-Aparicio C, Criado G, Pastor J, Dianzani U, Portolés P, Rojo JM ( 2016 ).ETP- 46321, a dual p110a/δ class IA phosphoinos- itide 3-kinase inhibitor modulates T lym- phocyte activation and collagen-induced arthritis. Biochem Pharmacol 106, 56-59. ∞ PATENT ∞ Pastor J. et al. ( 2016 ). New Compounds. PCT/GB2016/052641. ANNUAL REPORT 2016 168 DIRECTION OF INNOVATION CNIO - LILLY CELL SIGNALLING THERAPIES SECTION SCOPE OF THE ELI LILLY - CNIO PARTNERSHIP Eli Lilly and CNIO are collaborating on the identification and validation of novel targets in cancer immunometabolism. Our Section is funded through a research contract with Eli Lilly and focuses on the identification of small molecular weight molecules that regulate the metabolism of malignant cells, with the objective of killing them, either directly, acting synergistically with other anti-tumour agents, or activating the anti-tumour immune response. Exploring how to better target these mechanisms would lead to better and more efficient therapeutic options. A combination of in vitro and in vivo approaches is being utilised to obtain a complete understanding of the metabolic reprogramming regulated by oncogenes like RAS, as well as the characterisation of the metabolic status of tumours ( Cerezo A. et al, February 2016 ; Keystone symposium meeting on ‘ New Frontiers in Understanding Tumor Metabolism ’ in Banff, Canada ). For this purpose, we have developed a series of biochemical and cell-based assays exploiting advanced techniques such as extracellular flux analysis ( Seahorse technology ), NMR and metabolomics. Finally, each target goes through an in vivo validation process using xenografts, allografts and mouse models developed at the CNIO ; this process includes the use of non-invasive in vivo imaging technologies, as well as the immunohistochemical characterisation of tumours for different metabolic, immune and tumour markers. Susana Velasco Section Head Staff Scientists Ana Cerezo, Juan Manuel Funes ( since March ), Eva P. Lospitao, Gloria Martínez Del Hoyo Technicians Laura Diezma, Tamara Mondejar ( TS )*, Sandra Peregrina ( TS )*, Estela Casas López ( since December ) *Titulado Superior ( Advanced Degree ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 169 EXPERIMENTAL THERAPEUTICS PROGRAMME | CNIO - LILLY CELL SIGNALLING THERAPIES SECTION SCIENTIFIC CONTEXT The observation of an altered metabolic state in cancer cells dates back to the early 20th century when Otto Warburg observed that cancer cells preferentially utilise glycolysis over oxidative phosphorylation for growth, even in the presence of normal oxygen levels ( Warburg 1956 ), a phenomenon known as the ‘ Warburg effect ’. Warburg argued that, ‘ this altered metabolic state was the underlying cause for cancer ’. The molecular mechanisms driving an altered tumour metabolism have only recently begun to be understood as a result of large-scale genomic sequencing as well as advances in metabolomic profiling technologies. Recent studies have shown that many oncogenes, including Myc and Ras, impart an altered metabolic phenotype in cancer cells through the regulation of genes involved in central metabolic pathways such as glycolysis, fatty acid metabolism, oxidative phosphorylation, and the one carbon pool. Cellular metabolism is a fine tuned process ; tumours may rely heavily on specific metabolic pathways to obtain their energy while using other pathways to grow in order to give tumour cells a growth advantage. This situation may leave tumour cells in a frail position under certain treatments or circumstances, while normal cells may be able to compensate and survive ( FIGURE, C ). Furthermore, the high requirements of nutrients and other soluble factors, and the release of metabolites with immunosuppressive properties, together with the hypoxic conditions found in tumours, creates a ‘ non-friendly ’ microenvironment for an anti-tumour immune surveillance, while facilitating the growth of other tumour-promoting cells such as the stroma and myeloid cells ( FIGURE A,B ). Thus, the mechanistic understanding of cancer metabolism has led to renewed interest in developing therapeutics that target key enzymes involved in this process. Checkpoint- blockade immunotherapy has been one of the most exciting advances made in cancer treatment in recent years. Metabolic interplay in the local microenvironment can mediate T cell differentiation and function. ‘ Checkpoint-blockade ’ antibodies can also influence cellular metabolism. Finally, recent clinical trials have shown that combination immunotherapy based on immune checkpoints blockade, provides even higher response rates than either approach alone. s Figure ( A ) Immunohistochemical analysis of metabolic enzyme PKM2, showing the tumour microenvironment in a PANIN-3 from a Pancreatic Adenocarcinoma model Elas-tTA/tetO-Cre ;K-Ras(+/LSLG12Vgeo );P53( lox/lox ): PDAC ( Carmen Guerra & Mariano Barbacid ). In addition to the tumour cells, there are fibroblasts, endothelial, myeloid cells and immune cells that may be facilitating tumour growth, while the immune cells involved in the anti-tumour immune response are absent or inhibited. ( B ) Immunohistochemical analysis showing that the expression of the immune checkpoint ligand PDL1 correlates with the expression of specific tumour profiling markers ( SERBP1, Ambrogio et al., 2016 ) and certain metabolic markers ( LDHB and FASN ) in a lung adenocarcinoma derived from a KrasLSLG12Vgeo ;Trp53lox/lox ( Chiara Ambrogio & Mariano Barbacid ). ( C ) Cartoon depicting a strategy to control tumour growth through the regulation of specific metabolic targets. Tumours may rely heavily on specific metabolic pathways to grow and evade immune surveillance, as well as to obtain energy. This rapid growth also results in increased DNA damage, either through an increase in the production of ROS or due to replication stress. This situation leaves tumour cells more vulnerable to certain metabolic interventions as well as increasing the anti- tumour response of the immune system. ANNUAL REPORT 2016 170 DIRECTION OF INNOVATION CNIO - LILLY EPIGENETICS SECTION “ Our goal is to identify epigenetic events that contribute to tumourigenesis and that might be susceptible to modulation by therapeutic agents.” María José Barrero Section Head Staff Scientist Sergio Ruíz Technicians Verónica García ( TS )*, Jacinto Sarmentero ( TS )* *Titulado Superior ( Advanced Degree ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 171 EXPERIMENTAL THERAPEUTICS PROGRAMME | CNIO - LILLY EPIGENETICS SECTION SCOPE OF THE CNIO - ELI LILLY PARTNERSHIP Eli Lilly and CNIO are collaborating on the identification and validation of novel targets in cancer epigenetics. Our Section is funded through a research contract with Eli Lilly and focuses on the identification of small molecular weight molecules that are able to modulate the epigenome of malignant cells, and ultimately block the growth and spread of tumours. Potential targets ( FIGURE ) are being validated in vitro and in vivo using animal models developed at the CNIO. Furthermore, we are currently setting up biochemical and cell-based assays with the aim of understanding the mechanism of action of such targets at the molecular level. SCIENTIFIC CONTEXT Recent studies have shown that the alterations that take place in cancer cells not only occur at the DNA sequence but also at the level of the epigenome. Eukaryotic DNA is wrapped around histone proteins to constitute chromatin, which plays fundamental structural and regulatory roles. The epigenome consists of chemical changes in both DNA and histones that can be inherited through cell division and are controlled by the action of a large set of epigenetic regulators that possess enzymatic activity. Ultimately, DNA and histone modifications control the level of chromatin condensation, which in turn regulates the accessibility of transcription factors to the chromatin and, therefore, gene expression. During the past few years several studies, including our own, have suggested that the deregulation of the chromatin- modifying machineries can lead to aberrant gene expression causing cancer and other human diseases. The epigenome is regulated in a highly dynamic fashion by the coordinated action of regulators that are able to write, erase and read histone and DNA modifications ( FIGURE ). Thus, contrary to genetic mutations, epigenetic aberrations can be reversed by targeting the appropriate epigenetic regulators. Indeed, drugs targeting DNA methyltransferases and histone deacetylases have successfully demonstrated anticancer properties and are currently used in the clinic. Therefore, identifying the molecular function of critical epigenetic regulators and their complex relationship with the cancer epigenome, as well as the development of small molecular inhibitors of their activities, hold great promise for cancer therapy ( FIGURE ). s ∞ PUBLICATIONS ∞ García-Carpizo V, Sarmentero J, Han B, Graña O, Ruiz-Llorente S, Pisano DG, Ser- rano M, Brooks HB, Campbell RM, Barrero MJ ( 2016 ). NSD2 contributes to oncogenic RAS-driven transcription in lung cancer cells through long-range epigenetic ac- tivation. Sci Rep 6, 32952. ∞ Castaño J, Morera C, Sesé B, Boue S, Bonet-Costa C, Martí M, Roque A, Jordan A, Barrero MJ ( 2016 ). SETD7 Regulates the Differentiation of Human Embryonic Stem Cells. PLoS One 11, e0149502. Figure Strategies for targeting epigenetic regulators. The enzymatic activities of DNA methyltransferases ( DNMTs ), protein lysine methyltransferases ( PKMTs ), protein arginine methyltransferases ( PRMTs ), histone acetyltransferases ( HATs ), histone deacetylases ( HDACs ), or lysine demethylases ( KDMs ), are amenable to inhibition by small molecules. Additionally, molecular probes can be used to block the interactions of readers containing PHD, Bromo, Chromo or Tudor domains with modified histones, or to disrupt the interaction between critical core components of chromatin-related complexes. ANNUAL REPORT 2016 172 DIRECTION OF INNOVATION TECHNOLOGY TRANSFER AND VALORISATION OFFICE Anabel Sanz Director Technology Transfer Manager M. Cruz Marín Technician Rocío Manzano ( Pej-L )* *Plan de Empleo Joven- Licenciado ( Youth Employment Plan-Graduate ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 173 TECHNOLOGY TRANSFER AND VALORISATION OFFICE The CNIO’s Technology Transfer and Valorisation Office at CNIO ( TTVO ) acts as a bridge that connects the research results generated by CNIO’s scientists to the Centre’s commercial partners, thereby helping to ensure that new products are developed for the public interest. Without this bridge the public, and in particular cancer patients, would not benefit from the full potential of the discoveries made by CNIO scientist. Close alignment of the technology transfer expertise with CNIO’s innovation strategy priorities serves to optimise the transfer of the research materials and novel scientific discoveries to the health sector. The TTVO performs an all-round management and follow-up of all aspects, including relationships with stakeholders, in order to ensure the appropriate intellectual property protection and commercial viability of the research results generated by CNIO’s scientist. Additionally, the Office proactively follows up on the progress of scientific activity at the CNIO in order to identify projects with high transfer potential ; it plays a managerial and advisory role in the entire process in order to ensure the efficient use of the patent system, identify appropriate commercial partners for a timely development of technologies, negotiate licenses, monitor the activities of licensees regarding the achievement of milestones, and the payment of royalty fees. In this context, the TTVO Office managed the execution of 182 agreements related to the CNIO’s intellectual property in 2016. About 70% of these were of international nature, which is a reflection of the international relevance of the scientific research at the CNIO. Many pertain to the exchange of research materials and data with the external scientific community and health based companies. Other involved Research Collaboration Agreements ( RCAs ). RCAs are an important mechanism used by CNIO for successful cooperation with industry ; they ensure that the risks and benefits related to research results and intellectual property are shared. In 2016, the industry investment secured via RCAs totalled 3.8 Mio euros, nearly 10% of CNIO’s yearly budget. Public-private partnerships are potent tools for the valorisation of research results whereby scientific knowledge gets converted into diagnostic and therapeutic products and services. Valorisation of CNIO’s research results through alliances with industry is not just about ‘ money ’, but also about the impact that can be created for the public benefit and, in particular, for cancer patients. In 2016, a project based on the findings of CNIO scientists that boosting levels with nicotinamide riboside ( NR ) prevented and abolished aggressive tumour formation received nearly 1 Mio in funding from the MINECO Retos-Colaboración programme. The project was focussed on the development of a new NR-based therapy that could be used for Hepatocellular Carcinoma and other tumours. The inventions of CNIO scientists that have the potential to be transferred to the market are protected through patents. The CNIO’s patent portfolio is composed of 26 families. Licensed patents are managed by our licensees and the rest is managed by the TTVO Office. Patents and unpatented research tools are licensed to increase their availability to the scientific community, as well as to create opportunities for our business partners and to provide a financial return on public investment. The royalty fees collected from licenses in 2016 exceeded 650,000 euros. This income reverts back to CNIO research activities as well as to the inventors themselves. A total of 44 inventors, about 10% of CNIO’s researchers, have contributed and benefited from this achievement. Fostering an innovation culture among our scientists is one of our priorities. With the support of Fundación Banco Santander, we uphold our agreement with the prestigious IE Business School, through which many of our investigators − 3 new in 2016 − have already obtained training in market-oriented innovation strategies. Finally, these achievements stand testament to the excellence and hard work of the CNIO scientists and to CNIO’s unwavering encouragement of innovation and technology transfer activities. “ Valorisation of CNIO’s research results through alliances with industry is not just about ‘ money ’, but also about the impact that can be created for the public benefit and, in particular, for cancer patients.” ANNUAL REPORT 2016 174 DIRECTION OF INNOVATION PRIVATE SPONSORS “ We take this opportunity to express our thanks and appreciation to all our sponsors and donors for the generous support that we received from them in 2016. They play an inherent role in our present and future successes.” One of the Fundación “ la Caixa’s ” main goals is to support an innovative programme aimed at fostering international fellowships in order to attract the most outstanding students from the international arena to obtain their doctoral degrees at accredited ‘ Severo Ochoa ’ Centres of Excellence. This acclaimed “ la Caixa ”-Severo Ochoa International PhD Programme assures highly competitive standards by guiding exceptional students towards a career in oncology research ; a basic principle is that the selection process is not to be limited to Spanish students only but also includes international students. During 2016, 2 pre-doctoral students received one of these internationally recognised fellowships. The Fundación “ la Caixa ” also helps finance our most prominent international conferences, the CNIO-”la Caixa ” Foundation Frontiers Meetings. Fundación CRIS is dedicated t o t h e p r o m o t i o n a n d development of research with the aim of eliminating the serious health threat of cancer. Fundación CRIS generously supports 4 research groups at the CNIO : the Prostate Cancer Clinical Research Unit ( CRU ), headed by David Olmos ; the Breast Cancer CRU, headed by Miguel Quintela ; and the H12O-CNIO Haematological Malignancies CRU, led by Joaquín Martínez-López. These Groups focus on the translation of advances in cancer research into improvements in patient care. T h e F u n d a c i ó n S e v e Ballesteros is a private not- for-profit institution focused on securing, financing and promoting research projects centred on brain tumours. Fundación Seve Ballesteros supports the Seve Ballesteros Foundation – CNIO Brain Tumour Group, headed by Massimo Squatrito, since 2012. This Group focuses on the identification of markers for brain tumours as its principal activity. The Fundación Banco Santander funds the Banco Santander Foundation – CNIO Fellowships for Young Researchers. These fellowships are aimed to support highly talented and motivated young scientists who trained in the UK/USA and wish to pursue their postdoctoral training at the CNIO. One young scientist, Ana Ortega, who came from the Sloan Kettering Institute for Cancer Research in New York, joined the CNIO thanks to a Banco Santader Foundation-CNIO Fellowship in 2016. Additionally, thanks to the support of the Fundación Banco Santander, a group of 3 young researchers received training on managerial and entrepreneurial skills, in collaboration with the IE Business School. The Fundación Marcelino Botín and the Banco Santander are committed to supporting scientific research and knowledge transfer from academia to the market through science programmes ; this transfer is regarded as one of the main driving forces for Spain’s economic and social development. These 2 well-recognised organisations collaborate with the CNIO in this regard by supporting the research groups led by Manuel Serrano, Maria A. Blasco and Óscar Fernández-Capetillo. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 175 OTHER SPONSORS The Centre also benefits from generous support from private institutions, companies and foundations, as well as via external fundraising from local associations that are equally dedicated to the battle against cancer : Fundación Juegaterapia, Fundación Investigación Biomédica Hospital Universitario 12 de Octubre, Fundación Banco Sabadell, Asociación Española Afectados por Sarcoma ( AEAS ), Fressia Group, Compañia Logística de Hidrocarburos, Asociación para la Investigación y Formación Neoplásica ( ASIFEN ), ASISA Vida, French Embassy , AVON Cosmetics S.A. And last but not least, we would also like to extend our heartfelt thanks to all ‘ CNIO Friends ’ donors, sponsors and benefactors who − with their generous donations to support cancer research at the CNIO − have ensured the continuation of our research endeavours throughout 2016. PRIVATE SPONSORS T h e F u n d a c i ó n J e s ú s Serra-Catalana Occidente continues to fund the Visiting Scientists Programme that was established to support prestigious international professors for short stays at the CNIO. The beneficiary of the Jesús Serra Foundation’s Visiting Scientist Award in 2016 was Patrick Sung, Professor of Molecular Biophysics and Biochemistry and of Therapeutic Radiology, Yale University School of Medicine in New Haven ( USA ). AXA Research Fund ( ARF ), a global initiative of scientific philanthropy run by the insurance group AXA, awarded an AXA-CNIO Endowed Permanent Chair in Molecular Oncology to Mariano Barbacid as part of its 2011 call. AMIGOS DEL CNIO más INVESTIGACIÓN menos CÁNCER ANNUAL REPORT 2016 176 Communication SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 177 ANNUAL REPORT 2016 178 COMMUNICATION COMMUNICATIONS NURIA NORIEGA Head Of Communications Communications Officer (until April) Vanessa Pombo Communications Officer Cristina de Martos ( since April ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 179 This year, yet again, CNIO’s discoveries and activities were prominently featured in various domestic and international media outlets. In the printed and digital press the CNIO exceeded 2,600 media mentions ; it is clear that the CNIO is being brought closer to the public eye, thereby increasing awareness of cancer research and slowly interweaving it into our national culture. One of the most widely commented articles in 2016, published in the prestigious Science journal in November, concerned cell reprogramming ; it was published under the title Tissue Damage and Senescence Provide Critical Signals for Cellular Reprogramming in vivo, and was authored by the researchers Manuel Serrano and Lluc Mosteiro, from the CNIO Tumour Suppression Group. The discovery, which places the focus on tissue damage as a tool for cellular reprogramming and for the potential regeneration of tissue, was featured in major media outlets, including the front page of the daily El Mundo, as well as receiving radio and TV coverage. A major milestone in 2016 was the agreement that CNIO established with the Spanish Radio and Television Corporation ( RTVE ) on the occasion of the World Cancer Day, which took place on 4 February. Thanks to this collaboration, the Spanish public TV and radio broadcasting stations featured the voices of CNIO researchers and the latest developments in the oncology field. As one of the top leading Cancer Research Centres in the world, the CNIO also hit the headlines beyond our borders. In 2016, the CNIO submitted 26 press releases to the global news service, EurekAlert ! Throughout the year, these stories received nearly 83,000 hits from around the world and were taken up by prestigious international media such as the BBC, The Guardian, The Scientist or Scientific American. “ At the CNIO, we want to give science and research the strong voice they deserve as they are our true means of coping with cancer.” Our social networks are consolidating their communities. By December 2016, our Twitter channel had over 10,842 followers, with whom we keep an ongoing and valuable dialogue via the platform. The ‘ CNIO Friends ’ social media has also become more consolidated ; in December 2016, our Facebook page dedicated to this philanthropic initiative reached over 33,800 followers, highlighting, once again, the solidarity of our society and its growing interest in cancer research and the advances that can help fight the battle against cancer. It’s another joy for us to mention that in May, the ‘ CNIO Friends ’ Facebook page received the Internet Day Award in its category. A distinction that, within the short period of the initiative’s existence, has served to boost its wider public dissemination. ANNUAL REPORT 2016 180 COMMUNICATION SOCIAL MEDIA PLATFORMS CNIO FRIENDS CNIO FACEBOOK 33,837 FOLLOWERS TWITTER 10,842 FOLLOWERS TWITTER 567 FOLLOWERS YOUTUBE 12,370 VIEWS 3 % < 24 15 % < 24 14 % < 24 22 % < 24 13 % > 65 <1 % > 65 1 % > 65 9 % > 65 10 % 25-34 39 % 25-34 31 % 25-34 25 % 25-34 48 % 35-54 44 % 35-54 50 % 35-54 34 % 35-54 26 % 55-64 2 % 55-64 4 % 55-64 10 % 55-64 AGE DISTRIBUTION AGE DISTRIBUTION AGE DISTRIBUTION AGE DISTRIBUTION GENDER DISTRIBUTION GENDER DISTRIBUTION GENDER DISTRIBUTION GENDER DISTRIBUTION MALE MALE MALE MALE 17 % 46 % 38 % 58 % FEMALE FEMALE FEMALE FEMALE 83 % 54 % 62 % 42 % SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 181 PRESS CLIPPINGS 1 ABC, January 23, 2016 2 Expansión ( front page ), February 11, 2016 3 El Ojo Clínico, La 2, March 13, 2016 4 BBC, March 16, 2016 5 SINC, March 16, 2016 6 La Razón, March 27, 2016 7 Diario Médico, April 11, 2016 8 La Sexta Noticias, April 17, 2016 Investigaci ón BELÉN DÍA Z ALONSO de se espec ializó en el establecimi ento tem- prano de lo s circuitos n euronales d urante el desarrollo. « La mezcla d e la neuroci encia con la biología d el cáncer no s ha permit ido hacer cosas muy n ovedosas y abre grande s expec- tativas», ex plica Valien te. Su trabajó j unto a Mass agué aclaró cues- tiones que hasta enton ces se desc onocían. «Cómo sale una célula de un tumo r, cómo se infiltra en e l torrente s anguíneo y logra so- brevivir ya se sabía. E l enigma em pezaba cuando esa célula mali gna llega a u n capilar del cerebro, que es un va so sanguíne o tan fino que literalm ente queda atrapada». En este primer pas o mueren la mayoría de las célu- las metastá sicas que ll egan órgan o rector. Pero no es e l único obst áculo que d eben su- PILAR QUI JADA C on una bril lante carre ra en de- sarrollo en E stados Unid os, Ma- nuel Valien te (Zaragoz a, 1980) decidió volv er para dirig ir el pri- mer laborat orio dedicad o en ex- clusiva al e studio de la s metás- tasis cerebr ales en Espa ña. Empezó en mar- zo del año p asado, en el Centro Nac ional de Investigacio nes Oncológ icas. Trae co mo equi- paje lo apre ndido dura nte cinco a ños en el Memorial S loan Ketter ing Cancer Center de Nueva York , al lado del español Joa n Massa- gué, líder in ternaciona l en la inves tigación de metásta sis. A Nuev a York llegó desde el Instituto de Neurocienc ias de Alica nte, don- Manuel Vali ente, del CN IO, experto en el sistema ner vioso y el de sarrollo de m etástasis, investiga có mo una célu la de la mam a o el pulmón log ra multiplic arse en el ce rebro ONCOLOGÍ A La biología del cáncer y la neurocien cia abren grandes exp ectativas perar. Desp ués han de cruzar la ba rrera he- matoencefá lica, una rig urosa aduan a que ro- dea al cereb ro, formada por un «co rdón de seguridad» de células e specializada s que im- piden la ent rada de elem entos extra ños des- de la sangre . Otro interr ogante: cóm o pueden sobrevivir e sas células en un ambi ente tan diferente y e specializado como el del cerebro. Su trabajo c on Massagu é lo aclaró e n par- te. «De la mi noría de célu las canceríg enas que cruzan esa b arrera física , la mayoría mueren, porque no s aben cómo crecer en un ambien- te tan difere nte. Pero alg unas prospe ran gra- cias a una e specie de es cudo que la s protege de los meca nismos de d efensa del c erebro», señala con u n lenguaje m uy coloquial . Se acos- tumbró en Nueva York a explicar s us inves- tigaciones d e forma com prensible a los inte- grantes de las fundaci ones privad as que fi- nanciaban su trabajo. Mecanismo s de defens a «Las célula s canceríge nas deben a lterar la pared de los vasos sang uíneos para "asomar la cabeza" y entrar en e l cerebro. Y cuando cruzan esta barrera ha n de perma necer ad- heridas a la parte exter ior de los va sos san- guíneos par a sobrevivir . Estos proc esos de- penden de g enes que ent ran en funci onamien- to tanto ant es como des pués de que la célula tumoral lleg ue a este ór gano vital», explica. En el cerebr o hay célula s que se pon en en guardia cua ndo detecta n esos mov imientos extraños. E ntre ellas, lo s astrocitos , «encar- gados de pro teger a las n euronas y d e alimen- tarlas, y qu e, por tanto juegan un papel im- portante. So n capaces d e detectar la presen- cia de una ú nica célula cancerígena ». Como su no mbre indica , los astrocit os tie- nen forma d e estrella, co n muchas r amifica- ciones para entrar en co ntacto con n euronas, vasos sangu íneos y otro s astrocitos . Cuando algo va mal , se vuelven reactivos, e s decir, se transforma n físicamen te, como los superhé- Manuel Valiente en su laboratorio del CNIO 16 SALUD abc.es/salu d SÁBADO , 23 DE ENE RO DE 2016 ABC Jueves 11 febrero 2016 43Expansión DIRECTIVOS CRUCERO Hurtigruten lanza nuevos itinerarios de cruceros con la cultura noruega como fondo. La música se convertirá en protagonista en el recorrido Rumbo Norte y Rumbo Sur, con los acordes de las obras del compositor Edward Grieg y excursiones en tierra con conciertos en cuevas, iglesias locales o antiguas fábricas de pescado. LIBRO Ayer llegó a las librerías Historia de un canalla (Plaza & Janés), la última novela de Julia Navarro. Las confesiones del publicista y asesor de imagen Thomas Spencer diseccionan una época, desde los años 80 hasta la actualidad, entre Londres y Nueva York. Navarro ha vendido cinco millones de ejemplares en más de 30 países de sus cinco libros. DESTINOS Córdoba es la ciudad más reservada en invierno, según el portal Hoteling. Los españoles se decantan por destinos nacionales y grandes ciudades para los días más fríos del año. A la ciudad andaluza le siguen Madrid, París, Londres y Sevilla. PARA DESCONECTAR Los nuevos itinerarios de Hurtigruten están ligados a la cultura noruega. Córdoba, la ciudad más reservada en invierno. “Ha llegado la hora de recuperar talento” MARÍA BLASCO Directora del Centro Nacional de Investigaciones Oncológicas (CNIO). Emelia Viaña. Madrid “¿Cómo compagina la direc- ción del Centro Nacional de Investigaciones Oncológicas (CNIO) con su carrera como investigadora?” Así empieza la entrevista con María Blasco y no podría haber elegido una pregunta peor. “Si fuera un hombre nunca me harían esa pregunta. Creo que a Mariano Barbacid –que dirigió la institu- ción antes que ella– nunca se la hicieron”, señala Blasco, que cree que uno de los problemas para que la mujer no esté en puestos de responsabilidad son los estereotipos. “Hay sesgos inconscientes que pesan nega- tivamente en la visión que la so- ciedad tiene del liderazgo de una mujer. La pregunta que me has hecho es uno de ellos”. Consciente o inconsciente- mente, lo cierto es que a las mu- jeres se les pregunta en muchas entrevistas por familia y trabajo mientras que con los hombres sólo se habla de trabajo. Tras el error –en mi caso, inconscien- te–, Blasco hace un alegato a fa- vor de ellas: “El 80% de los in- vestigadores en un laboratorio son mujeres, pero el 80% de los cargos de responsabilidad está en manos de hombres”, señala Blasco, que asume que el CNIO no está a salvo de este mal gene- ralizado. “Es un problema que me preocupa mucho, por eso hemos impulsado iniciativas para corregirlo. La jornada con- tinua o la obligatoriedad de no poner reuniones a las siete de la tarde van encaminadas a solu- cionarlo. Además, estamos ayu- dando a las mujeres a aumentar su red de contactos porque mu- chos puestos de responsabili- dad se otorgan porque conoces cómo trabaja alguien y necesi- tan ganar visibilidad”, explica Blasco, que participa en el pro- grama L’Oréal-Unesco For Wo- men in Science. El discurso de Blasco no nace del rencor, de hecho ella es una de esas mujeres que ha llegado a lo más alto en su carrera pro- fesional. ¿Cree que le ha costa- do más por ser mujer? “Me he tenido que esforzar el doble que un hombre para ver reconocido mi trabajo. Yo lo he conseguido, pero eso no quiere decir nada”. Buena parte del trabajo de Blasco fuera del laboratorio –está especializada en telóme- ros y telomerasa y su función en el cáncer y el envejecimiento, y fue fundadora de la empresa de biotecnología Life Length– tie- ne que ver con la política. De hecho, su jefe directo es otra mujer, Carmen Vela, secretaria de Estado de Investigación, De- sarrollo e Innovación. En el pa- tronato del CNIO están repre- sentadas algunas instituciones públicas y sus miembros cam- bian cada vez que se celebran elecciones, algo que Blasco acepta, pero que le parece sor- prendente si lo compara con instituciones de EEUU –traba- jó en el laboratorio Cold Spring Harbor de Nueva York, en el equipo de Carol Greider, pre- mio Nobel de Medicina en 2009 por un trabajo que inició cuando Blasco trabajaba allí–. De EEUU, Blasco ha impor- tado la necesidad de que cen- tros como el CNIO dispongan del apoyo privado. Las funda- ciones de La Caixa, BBVA o Santander, así como Iberia o Axa ya son patrocinadores de la institución, que cuentan con la colaboración de Servired o Hewlett Packard y el apoyo de particulares a través de la ini- ciativa Amigos del CNIO. “La fi- lantropía individual es algo po- co habitual en España. La socie- dad debe sentirse parte de la in- vestigación”, señala Blasco, que cree que éste es uno de los pro- blemas más graves de la ciencia en España. “Si conseguimos que las tres patas de la ciencia –investigación, innovación y aplicación clínica– engrasen, podremos explicar su rentabili- dad”. El CNIO es uno de los po- cos centros que patenta fárma- cos nacidos en su laboratorio gracias a acuerdos con farma- céuticas, algo que le permite obtener retornos económicos. Blasco dirige el centro desde 2011 y ha tenido que hacer re- cortes. Gracias a eso, la investi- gadora se ha visto obligada a pensar en éstas y otras formas de financiación que ahora dan sus frutos. “Gracias a esos retor- nos económicos estamos em- pezando a contratar a investi- gadores españoles en el extran- jero y que han adquirido una formación fundamental. Ha lle- gado la hora de recuperar ese talento”, concluye Blasco, que aclara que la formación en el extranjero no tiene que trasla- darse a la sociedad como nega- tivo. “La mitad de nuestros es- tudiantes son extranjeros y no sorprende en sus países. El CNIO es un polo de atracción de talento y uno de los retos es que lo siga siendo en el futuro”. Blasco dirige el CNIO desde 2011 y se formó con Margarita Salas, Mariano Barbacid o la Premio Nobel Carol Greider. Está especializada en telómeros y telomerasa. Blasco ha impulsado iniciativas como ‘Amigos del CNIO’ y ha buscado el apoyo de empresas para acercar la ciencia a la sociedad Fundó la empresa de biotecnología Life Length y defiende que la ciencia dé retornos económicos “Durante mi carrera, me he tenido que esforzar el doble que un hombre para ver reconocido mi trabajo” JM C ad en as SALUD /INVESTIGACIÓN que se le ha extirpado quirúrgicamente, es decir, ese tejido que sobra una vez rea- lizado el correspondiente diagnóstico por parte de los especialistas en Anatomía Patológica, y que en circunstancias nor- males se destruiría. Y es que ese exceden- te puede ser de gran importancia para investigación. «No le va a suponer ningún nuevo procedimiento ni molestia al donan- te», aclara. En caso de que el individuo quiera donar, también puede preguntar directamente en los grandes hospitales si tienen biobanco para hacerlo. LAS MUESTRAS¿Qué se hace después con esas muestras? Algunas se incluyen en parafi na y se conservan a temperatura ambiente, mientras que otras se congelan y se man-tienen a menos 80 grados o a menos 190 grados, depen-diendo del tipo de muestra. «Se congela en tiempo muy corto tras obtenerlas, en menos de dos horas», explica Matías Guiu. «En cuanto a la sangre, se fracciona y se consigue la máxima diversidad de produc- to para dar mayor rendimiento», matiza Morente. Pero, además, los biobancos continúan en evolución y los tipos de muestras varían. «Por ejemplo, en cáncer, se almacenan muestras de tejidos (tisulares) pero a veces hay tumores poco accesibles para tomar muestras, por lo que ahora se trabaja en una estrategia para obtener el DNA que las células tumorales vierten a la sangre. Es un nuevo tipo de muestra llamada biopsia líquida», explica el director del Biobanco IRBLleida. SENCILLO A la hora de ceder las muestras a investi- gadores, el procedimiento es sencillo. «Primero un Comité de Ética de Investiga- ción debe aprobar el proyecto que quieren llevar a cabo. Una vez dado este paso, se muestras de tejidos de pacientes, 135.000 de las cuales han sido aprobadas en el último año para ser usadas en más de 1.300 pro- yectos de investigación. «La Plataforma sirve para que estemos todos en red y po- damos vehiculizar las solicitudes de toda España», explica Xavier Matías Guiu, di- rector del Biobanco IRBLleida.Es más, el grado de organización colec- tiva en España es único en el mundo. «Fuimos los primeros en crear una red de biobancos con un modelo que está siendo copiado en Europa, América Latina y Oriente Medio», destaca Morente. Sin embargo, so-mos el único gran país de la Unión Europea que no está adherido a la Red Europea de Biobancos (Bbmri por sus siglas en inglés: Biobanking and Biomolecular Resources Research Infrastructures). « No se entiende que España, el noveno país en producción científica del mundo, no participe en esta estructura pan-europea especialmente cuando ha sido la pionera en estructuras cooperativas de biobancos. Esto nos resta presencia inter- nacional y difi culta al máximo el acceso a grandes proyectos internacionales, con su consiguiente pérdida científi ca y de retor- no de inversiones», argumenta Morente. Por su parte, Enrique de Álava, explica que «la pega, el presupuesto. Nuestra plataforma no está representada en las instituciones europeas. Hay una red euro- pea y nuestro gobierno no paga la cuota y eso nos corta las alas. Es un reto que nos conozcan». Lo importante es la labor al- truista, ya que son establecimientos (pú- blicos o privados) sin ánimo de lucro. En ellos se recogen muestras, no sólo de en- fermos, sino también de personas sanas que deseen colaborar, ya que «los necesi- tamos como controles», asegura el citado experto. En los hospitales, el biobanco «suele proponer al paciente que done parte de lo S on los llamados «bancos bue-nos», porque no cobran comi-siones y dan la mayor rentabi-lidad: investigar contra graves enfermedades. Y lo hacen al-macenando muestras biológicas humanas que ponen a disposición de la ciencia. Cerebros, tejidos, sangre, lágrimas, saliva… Todo con el objetivo de acabar con esas dolencias que todavía se nos escapan, como algunas cardiovasculares y neurológicas, tumorales, patologías infantiles o enfermedades raras. «La función de los biobancos es captar mues-tras humanas con datos asociados para que puedan ser utilizadas por investiga-dores en proyectos viables y de excelen-cia», explica Manuel Morente, director científi co del Biobanco del Centro Nacio-nal de Investigaciones Oncológicas (CNIO) y coordinador de la Plataforma Nacional de Biobancos. «Es una herramienta de apoyo a la in-vestigación. Se ceden y guardan muestras de pacientes, tanto enfermos como sanos y los datos clínicos se manejan con la supervisión legal y ética necesaria para que los investigadores realicen estudios», añade Enrique de Álava, jefe del Servicio de Anatomía Patológica del Hospital Universitario Virgen del Rocío, y miembro del comité de dirección de la Plataforma Nacional de Biobancos. POR QUÉ HUMANASPese a que las muestras que normalmente se utilizan de animales han sido y son muy útiles, no dejan de tener grandes di- ferencias con el ser humano, algo que co- rrobora Morente: «Una gran parte de la investigación se realiza utilizando mode- los, es decir, animales de experimentación y cultivos celulares. Pero, aunque estos modelos tradicionales tienen un valor tremendo, no son totalmente válidos debi- do a las grandes diferencias que tienen con los humanos. Esa diferencia es la que A. JIMENEZ  MADRID EJEMPLO DE SU GRAN UTILIDAD Un nuevo estudio liderado por el CNIO demuCientífi cos del Centro Nacional de Investigaciones Oncológicas ha demostrado que, en respuesta a un exceso de nutrientes, la proteína MCRS1 sirve de «interruptor» de otra proteína (mTOR) que está alterada en cáncer, diabetes y desórdenes asociados al envejecimiento. Los resultados del estudio que realizaron indican que el bloqueo de esta proteína podría ser efi caz para el tratamiento de este tipo de enfermedades. Las muestras para esta investigación fueron gestionadas a través del biobanco del CNIO. El trabajo fue publicado en la revista científi ca «Developmental Cell» y abre una vía al posible desarrollo de agentes bloqueantes de MCRS1 para poder tratar tumores y diabetes, fundamentalmente. Casi un centenar de centros guarda muestras de sangre, saliva, cerebros, mucosas, lágrimas... útiles para proyectos de investigación contra enfermedades graves. España fue el primer país que creó una red de biobancos con un modelo que, en la actualidad, se está copiando en Europa, América Latina y Oriente Medio Bi bancosAlmacenes de tejidos humanosal servicio de la ciencia La plataforma alberga dos millones de muestras, 135.000 han sido aprobadas en el último año para estudios vienen a salvar los biobancos, poniendo a disposición de los investigadores las muestras humanas correspondientes a pacientes reales», añade. «MADE IN SPAIN» En España, la mayoría se encuentran en hospitales, aunque también los hay en grandes centros de investigación. En la actualidad, existen unos 90 biobancos en nuestro país, 52 de los cuales trabajan dentro de la llamada Plataforma Nacional de Biobancos, una iniciativa del Instituto de Salud Carlos III (ISCIII) coordinada por el Biobanco del CNIO (Centro Nacional de Investigaciones Oncológicas). En ella, se reúnen en total cerca de dos millones de D O M I N G O . 2 7 d e M A R Z O d e 2 0 1 6 • 10 • 1 2 3 4 5 6 8 "La secuenciaci ón genómi- ca es la tecnolog ía que evo- luciona más ráp idamente, influye directam ente en casi todas las discip linas de la biología y la bi omedicina, y cada vez tiene mayor im- pacto social. Per o es que en el futuro se volv erá ubicua: dispondremos de secuen- cias para todo g racias a su integración en d ispositivos móviles de inform ación que, por ejemplo, pos ibilitarán la monitorización continua de nuestro estado d e salud". El vaticinio de A lfonso Va- lencia, presiden te de la So- ciedad Internaci onal de Bio- logía Computac ional (ISCB, por sus siglas e n inglés) y director del Pr ograma de Biología Estruc tural y Bio- computación e n el Centro Nacional de Inve stigaciones Oncológicas (C NIO), pasa por instrumen tos de se- cuenciación má s pequeños, baratos y veloce s, gracias al avance vertigi noso de la bioinformática y la biología computacional . Dos disci- plinas entrelaz adas e im- prescindibles pa ra el "man- tenimiento, org anización, accesibilidad y t ransforma- ción en conocim iento de la información gen ómica". Fue la materia d e debate La bioinformática a vanza en el estudio del ge noma La integración de la genó mica en dispositivos móv iles la volverá ubicua, según los expertos en este campo BARCELONA ANTONI M IXOY dmredaccion@ diariomedico.co m para los 185 in vestigado- res (de 32 paíse s) que par- ticiparon en la c onferencia internacional N ext Genera- tion Sequencin g (NGS), or- ganizada por e l ISCB y el Centro de Regul ación Genó- mica (CRG) y ce lebrada por segundo año con secutivo en Barcelona. La llamada zon a oscura del genoma fue uno de los principales tem as de deba- te. El exoma es e l 1 por cien- to del genoma q ue codifica para proteínas y está rela- cionado con la mayor par- te de funciones y mutacio- nes conocidas, y así, con la predisposición a enferme- dades. Por el co ntrario, la información ge nómica del 99 por ciento r estante es muy poco cono cida y difí- cil de explorar. "P ero no está ahí para nada, porque hoy sabemos que b uena parte debe de tener fu nciones". Se especula con la posible existencia en es ta inmensa zona no codifica nte de inte- rruptores que ha gan que en cada órgano s e expresen unos genes y no otros. "Así, una mutación en uno de es- tos reguladores -por ejem- plo, en el desarro llo cerebral fetal- podría res ultar fatídi- ca". Por ello se trabaja en nuevos método s computa- cionales y expe rimentales para el estudio de estas re- giones. Frente a la visió n estática del genoma (tene mos el mis- mo toda nuestr a vida), en los últimos dos a ños ha pro- gresado el análi sis de cómo responde a los es tímulos del entorno. "Para e llo secuen- ciamos los pro ductos del genoma activad o, los ARN", que ofrecen in formación más directa so bre el esta- do funcional d e los siste- mas. En el NGS se debatió sobre la impla ntación de tecnologías de secuencia- ción de ARN en la práctica clínica. En este vínculo entre ge- nómica y ento rno clínico destacó la prese ntación de Martin Reese, q ue expuso los sistemas de análisis de mutaciones den tro del Pro- yecto 100.000 G enomas, en marcha desde 2014, que prevé la integra ción de esa información en el sistema de salud del Rein o Unido. El objetivo es a ele rar el esta- blecimiento de d iagnósticos y tratamientos en función del genoma clín ico. Tras la secuenciación m asiva, orga- nizada de modo industrial, el análisis corre a cargo de la compañía Om icia -vence- dora en un conc urso inter- nacional-, "que l ogra pasar, en tan solo 36 h oras, de las secuencias a in formación clínicamente re levante". Alfonso Valenci a, director del P rograma de Bio logía Estructura l y Biocomputac ión en el CNIO. J A U M E C O S IA L L S 11/04/2016 Tirada: Difusión: Audiencia: 39.316 39.315 117.945 Categoría: Edición: Página: Rev Salud Nacional 23 AREA (cm2): 423,7 OCUPACIÓN: 39,6% V.PUB.: 1.752 SECTOR||NOTICIAS DE CNIO 7 ANNUAL REPORT 2016 182 COMMUNICATION 9 Lab24, 24h, May 10, 2016 10 La Razón, June 3, 2016 11 Correo Gallego, June 10, 2016 12 El Diario Vasco, June 13, 2016 13 Diario Médico ( front page ), July 11, 2016 14 ABC, July 16, 2016 15 Gaceta Médica, July 18, 2016 16 La Voz de Galicia, July 29, 2016 17 Diario Médico, September 14, 2016 18 ABC, September 24, 2016 CIENCIA &TECNOLOGÍA T elómero es la palabra. Si usted es asiduo a estas páginas de cien- cia la habrá leído en muchas ocasiones durante los últimos años. Sobre todo en informaciones relaciona- das con la senectud y el cáncer. Es uno de los santos griales de la investigación molecular contra las enfermedades derivadas de la vejez. Y, ahora, un espectacular avance del Centro Nacional de Investigaciones Oncológicas (CNIO) lo ha vuelto a poner de manifi esto. Un grupo de científi - cos dirigido por María Blasco ha logrado crear ratones con telóme- ros mucho más largos de lo nor- mal, sin necesidad de alterar sus genes. Son animales que mues- tran menos signos de envejeci- miento celular y menor propen- sión a padecer cáncer. Para expli- car por qué, habrá que empezar por el principio. Investigadores del CNIO crean ratones con telómeros hiperlargos a los que les afecta menos la vejez y la enfermedad EL SUEÑO DE ENVEJECER MÁS DESPACIO Y CON MENOS CÁNCERES LOS CIENTÍFICOS HAN LOGRADO ALARGAR LOS TELÓMEROS SIN NECESIDAD DE MODIFICAR LOS GENES Los telómeros son estructuras proteínicas que se encuentran en los extremos de todos los cromosomas y los protegen de la degradación. Se sabe que buena parte de su buen funcio- namiento depende de su longi- tud. Cada vez que una célula se divide tiene que copiar su mate- rial genético empaquetado en forma de cromosoma. Pero esa copia no es perfecta y los extre- mos de los telómeros pueden perderse, como si al fotocopiar un documento se quedara fuera la última letra de cada línea. Cuando el acortamiento progre- sivo de los telómeros llega a un grado crítico aparecen signos de muerte celular, de senectud y de algunas enfermedades. El organismo cuenta con una enzima, la telomerasa, que se encarga de reparar los telómeros. Pero su actividad también decae con el paso del tiempo. Hay dos tipos de célula, sin embargo, en las que la telomerasa no pierde nunca fuelle: las células cancero- sas y las células pluripontentes. En estos casos, los telómeros no se acortan. Por eso las células cancerosas son tan longevas y las pluripontentes (las llamadas «madre») tan activas. Lo malo es que la longitud de los telómeros viene determinada por la herencia genética. Cada especie de vertebrado tiene su propio rango de longitudes y en cada una de ellas el ritmo de acor- tamiento es diferente. Los teló- meros de los ratones, por ejem- plo, se acortan 100 veces más rá- pido que los humanos y los de los perros a una velocidad 10 veces mayor. De manera que no parecía que existiera forma de intervenir externamente en el acortamiento telomérico... hasta ahora. O, mejor dicho, hasta hace poco. Ya en 2009, el propio CNIO demostró que si se cultivan in vitro células iPS (células madre pluripontentes inducidas, es decir células adultas reconverti- das en madre) se podía forzar el alargamiento de los telómeros hasta lograr lo que los expertos llamaron «células con telómeros hiperlargos». Pero no era eviden- te que estas células pudieran ser capaces de desarrollarse hasta convertirse en un organismo complejo (como un ratón) con telómeros hiperlargos y que, por lo tanto, envejezca más despacio. Eso es exactamente lo que parece haber descubierto el trabajo anunciado ayer en Nature Com- munications y que ha sido fi rma- do por la doctora Elisa Varela como primera autora. Sí es posi- ble crear ratones con una longi- tud telomérica mayor sin necesi- dad de manipular sus genes. Hasta ahora la única estrategia para lograr este alargamiento inducido era modifi car en el animal los genes responsables de la producción de telomerasa. Pero los organismos ahora pre- sentados han mantenido la ar- quitectura genética necesaria para alargar los telómeros sim- plemente a través de la mo- difi cación de células pluri- potentes in vitro. Los anima- les en los que se empleó esta técnica mantuvieron la longitud extra de sus telómeros incluso 12 meses después de nacer y, lo más interesante, acumularon menos daños en su ADN, mostraron una mejor capacidad para reparar los daños genéticos acumulados y presentaron una menor inciden- cia de cánceres. Es decir, por primera vez se ha demostrado que se pueden ge- nerar tejidos capaces de afrontar mejor el paso del tiempo y de responder mejor a las enferme- dades que de él se derivan. En el futuro, el CNIO pretende generar ratones en los que todas sus célu- las cuenten con telómeros el do- ble de largos de lo normal. Sólo así se podrá responder a las pre- guntas que seguro que usted ya se está haciendo. ¿Estos ratones vivirán el doble? ¿Serán más lon- gevos y sanos? ¿Servirán para enseñarnos a producir tejidos sanos con los que podremos re- parar los tejidos envejecidos de nuestro cuerpo? La respuesta está en los telómeros. Jorge ALCALDE Director de «Quo» En verde, los telómeros hiperlargos; en rojo, los normales Tutankamón fue enterrado con una daga hecha de una plancha que vino, literalmen- te, del espacio, según análisis de la composición de la pe- queña espada de hierro halla- da en el sarcófago del faraón. Usando espectrometría de fl uorescencia de rayos X por- tátil no invasiva, un equipo de investigadores italianos y egipcios confi rmó que la daga de hierro colocada en el mus- lo derecho del cuerpo momi- fi cado de Tutankamón tiene origen meteórico. El equipo, que incluye in- vestigadores del Politécnico de Milán, Universidad de Pisa y el Museo Egipcio de El Cairo, detalla sus resultados en la revista «Meteoritics and Pla- netary Science». El arma, LA DAGA DE TUTANKAMÓN NO ERA DE ESTE MUNDO Por L. R. C. - Madrid ahora en exhibición en el Museo Egipcio de El Cairo, fue descrita en 1925 por Howard Carter, quien descubrió tres años antes la tumba llena de tesoros, como «una daga de oro muy ornamentado con un pomo de cristal». Hecho de metal no oxidado, la hoja está fi namente decorada con un mango de oro. Se comple- ta con una funda de oro ador- nada con un estampado de fl ores de lirio en un lado y plumas en el otro en el otro, rematada con la cabeza de un chacal. Las últimas mejoras tecno- lógicas han permitido a los investigadores determinar la composición de la hoja. «El hierro meteórico está clara- mente indicado por la presen- cia de un alto porcentaje de níquel», ha dicho la autora principal del estudio. Tutankamón y las dagas «extraterrestres» Viernes. 3 de junio de 2016 • LA RAZÓN 76 EL CORREO GALLEGO 45 VIERNES10 DE JUNIO DE 2016 TENDENCIAS La justicia europea obliga a España a revisar la compensación por copia privadaPROPIEDAD INTELECTUAL El Tribunal de Justicia de la UE tumbó ayer el sistema español de compensación por copia privada o canon digital, aprobado por el Gobierno del PP, al esti-mar que colisiona con la directiva comunitaria so-bre derechos de autor.La sentencia obligará a España a revisar este siste-ma aprobado en 2012 por el Ejecutivo de Rajoy. La vicepresidenta en funcio- nes, Soraya Sáenz de San-tamaría, aseguró ayer que trabajarán “a pleno rendi-miento” para hallar “una solución satisfactoria”.Las entidades de gestión de derechos de autor que iniciaron el proceso con-tra el actual canon –Egeda, Dama y Vegap– lo conside-ran “un espaldarazo a la cultura”, y recordaron que el Estado “tendrá que res-ponder por los perjuicios causados”, que calculan superiores a los cuatro-cientos millones de euros.La instancia judicial en-tiende que el sistema es-pañol “no asegura que el coste de la compensación equitativa sólo sea sufra-gado por los usuarios de copias privadas”. Y expli-ca que la compensación, con cargo a los presupues-tos generales del Estado, obliga a pagar a “todos los contribuyentes, incluidas las personas jurídicas”. EfE Desde el Ejecutivo“El Gobierno trabajará en una solución que sea satisfactoria para todos y plenamente cumplidora del derecho en la Unión Europea” Desde Adepi “Debe compensarse la enorme diferencia que existe entre el importe que se ha estado pagando y el que se debería haber pagado” La Unidad de Investigación Clínica de Cáncer de Mama del Centro Nacional de In-vestigaciones Oncológicas (CNIO) acaba de publicar un importante hallazgo re-lacionado con los antiangio-génicos, los fármacos más utilizados en el tratamiento del cáncer. En un paper pu-blicado en Cell Reports des-criben un mecanismo de resistencia a estos compues-tos y, lo que es más importan-te, una vía para combatirlo. ALbA LAgO Santiago Un hallazgo del CNIO paliará la resistencia al fármaco oncológico El mecanismo descrito abre vías para combatir el rechazo a los medicamentos antiangiocénicos Trabajando con ratones con cáncer de mama y pulmón, han observado que la adición de un antidiabético a los an-tiangiogénicos inhibe el cre-cimiento tumoral hasta en un 92 %. La resistencia a los fár-macos antiangiogénicos es un problema común en los tumores epiteliales y espe-cialmente preocupante si se tiene en cuenta que son de los agentes más utilizados en el tratamiento del cáncer, ya que se emplean contra los carcinomas de mama, pul-món, ovario, colorrectales, ••• El tratamiento con TKIs impide el metabolismo des- controlado de glucosa en las células tumorales, según Miguel Quintela-Fandino, jefe de la Unidad de Investiga- ción Clínica de Cáncer de Mama del CNIO, y su equipo. Esto, que debería provocar la muerte por inanición de las células cancerígenas, no siempre tiene el efecto letal de- seado. Muchos tumores resisten. Pero en este mecanis- mo los investigadores del CNIO han descubierto también una oportunidad para atacar a las células cancerígenas. adaptación riñón o hígado, entre otros.Los inhibidores de la tiro-sín quinasa (TKIs) forman parte de la familia de los an-tiangiogénicos. El ninteda-nib —un TKI con resultados hasta ahora superiores a los antiguos sorafenib o suniti-nib— acaba de recibir la apro-bación para el tratamiento del cáncer de pulmón avan-zado por la FDA y la EMEA. Por eso, “es importante defi-nir los mecanismos de resis-tencia adquirida a los TKIs”, según los autores del traba-jo. El desarrollo de los tumo-res está soportado por vasos sanguíneos anormales que causan una disminución del oxígeno en estos tejidos. La falta de oxígeno desencadena un cambio en el metabolismo celular. Es el conocido efecto Warburg, por el cual las cé-lulas tumorales consumen hasta 20 veces más cantidad de glucosa que las normales, prescindiendo así de los mo-tores habituales de las célu-las: las mitocondrias. Miguel Quintela-Fandino, María J. Bueno, Ivana Zagorac y Silvana Mourón Foto: CNIO Blasco, una de las voces autorizadas en la ciencia del envejecimi ento, habló en Biodo nostia de los avances en la investigación m olecular :: ARANTXA A LDAZ SAN SEBAST IÁN. La ciencia b us- ca desde hace a ños cómo retra sar el envejecimient o y alargar la vi da en condiciones san as, sin enferme dad. Que una empre sa como Google haya invertido cient os de millones de eu- ros en impulsa r proyectos de inves- tigación dirigi dos a encontra r fár- macos que perm itan ese elixir d e ju- ventud demue stra la importa ncia que ha adquiri do este campo de la investigación, q ue también tien e su ‘sucursal’ dono stiarra. El inst ituto de investigació n sanitaria Biod onos- tia estudia la fu nción de los tel óme- ros, los extrem os de los crom oso- mas que hacen de reloj de la v ida. La directora de l Centro Nacion al de Investigaciones Oncológicas (C NIO), María Blasco, h a visitado recie nte- mente el centro donostiarra par a ha- blar de los últim os avances. «Ll egar a los 70 años s ano parece est ar de- terminado sob re todo por el m odo de vida, y en m enor medida p or los genes», explica . – En el lago M aggiore, Italia , vive Emma Morano que, a sus 116 años y 118 días es la mujer más an ciana del planeta. H asta allí siguen yen- do científicos para saber cuá l es el secreto de su lo ngevidad. ¿Qué bus- can? – Una de las m aneras de ente nder por qué enveje cemos y cuále s son los genes que d eterminan este pro- ceso es estudia r a los centena rios y a los supercent enarios, como E mma Morano. Aunq ue estos estudi os sin duda son muy interesantes y han permitido avan zar, los avances más gún efecto inde seable, de hech o, los ratones vivían significativam ente mucho más. A hora lo que est amos haciendo es de mostrar que la telo- merasa puede servir para el trata- miento de enfe rmedades asoc iadas el envejecimie nto como cardi ovas- cular. Vimos qu e la telomerasa pro- tegía a los raton es de muerte p or in- farto de mioca rdio. – Empresas te cnológicas de la ta- lla de Google h an invertido g ran- des sumas de d inero en la cie ncia del envejecim iento. ¿Esto ay uda- rá a acelerar lo s resultados? ¿ Qué buscan exacta mente? – Google ha in vertido en una em- presa farmacéu tica que preten de ge- nerar un nuevo tipo de fármaco s que están inspirado s en el conocim ien- to de por qué e nvejecemos, e n las causas molecu lares del enve jeci- miento. La idea es que así se po drían tratar más efici entemente enf erme- dades que aho ra no tienen t rata- miento como l as neurodegen erati- vas... – Colabora co n Biodonostia , pri- mer instituto de investigació n sa- nitaria en Eus kadi. ¿Qué op ortu- nidades puede brindar un ho spi- tal para la inve stigación? ¿Se hace un tipo de inv estigación dif eren- te, más orient ada al pacient e? – Biodonostia e s un instituto j oven, con científicos y científicas jó venes que han estado investigando e n los mejores sitios del mundo. Ti enen ideas y proyect os muy romped ores, y necesitan tod o el apoyo econ ómi- co para llevarl os a cabo. No t engo duda de que si se les apoya, B iodo- nostia puede c onvertirse en u n re- ferente en la in vestigación bio mé- dica. Ya están haciendo muy bien algo que es fun damental y es que- rer y saber tras ladar los result ados a aplicaciones. – No puedo te rminar la entr evis- ta sin pregunt ar por la crisis en la investigación . ¿La recupera ción económica de la que se habl a lle- ga a la ciencia ? ¿O los recort es no tienen vuelta atrás? – Ha llegado p ero tímidame nte, y con mucha pr eocupación d e que podamos retro ceder de nuev o. Ha de haber una a puesta por la c ien- cia clara de to dos los partido s, ha de ser una prio ridad. Mientra s que los países líde res apuestan f uerte por la ciencia y la investiga ción como motor d el conocimien to y de la innovació n para asegura r- se un futuro c ompetiti- vo frente a tita nes como China, aquí se guimos teniendo que lu- char por una fi- nanciación suf i- ciente. «Los genes que nos mantienen jóve nes dejan de funcionar de forma óptima con el ti empo» «Conocer las rut as moleculares de l envejecimiento permitirá preve nir y tratar enfermed ades» «Seguimos preo cupados. Ha de haber una apuesta clara por la cien cia de todos los par tidos» importantes s obre el conocim ien- to de por qué e nvejecemos no han venido del estu dio de los supe rcen- tenarios sino de entender a nivel molecular qué determina la l onge- vidad en distin tos organismo s (gu- sanos, moscas, ratones...). Así, se han podido descub rir las rutas mo lecu- lares que son l as causantes d el en- vejecimiento y se ha conse guido manipular esta s rutas consigu iente adelantar o retr asar el envejeci mien- to en estos org anismos. Ente nder estar rutas mo leculares del en veje- cimiento tamb ién nos dará la s cla- ves para la prev ención y tratam ien- to eficaz de dis tintas enferme dades asociadas al en vejecimiento (cán- cer, cardiovascu lar, neurodegen era- ción...). – ¿Qué tienen de diferente la s per- sonas más lon gevas? ¿Mejor es ge- nes? – Los genes par ecen tener más peso que el modo de vida en aquella s per- sonas que llega n a ser centena rias o supercenterari as. De hecho, s e es- tudian familia s en las que m u- chos de sus mie mbros son cen- tenarios para en contrar si hay genes que det erminen la longevidad ex trema. Sin embargo, llega r a los 70 años sano pare ce estar determinado sobre todo por el mo do de vida, los genes tam- bién importan , pero en menor prop orción. Pero rec orde- mos que tanto los genes com o el modo de vida in ciden sobre los e ven- tos moleculare s que causan e l en- vejecimiento. – Recientemen te ha dicho qu e «el envejecimient o no está progr ama- do por la evolu ción, que no ha y cé- lulas Terminat or». Explíques e, por favor. – Lo que se pien sa es que no hay pro- gramas genéti cos ‘Terminato r’, es decir que no ha y programas ge néti- cos para hacer nos envejecer . Hay programas gen éticos para det ermi- nar que se form e el feto, para que crezcamos hast a la edad adulta , pero no hay program as genéticos par a en- vejecer. Lo que parece ocurrir es que los genes que n os mantienen jóve- nes, porque pro tegen del daño nues- tro material ge nético, dejan d e fun- cionar de man era óptima co n el tiempo y nues tras células ac umu- lan daño, lo cu al produce el en veje- cimiento de nu estras células y teji- dos, y eventua lmente, cuand o es- tas dejan de ha cer su función, la en- fermedad y la muerte. Las cé lulas madre, que so n las que regen eran nuestro organi smo, son partic ular- mente import antes ya que cu ando estas dejan de f uncionar por ac umu- lación de daño el organismo p ierde su capacidad d e autoreparaci ón. – En laborator io han demost rado que el enveje cimiento se p uede modular (ade lantar o retra sar). ¿Para cuándo r esultados en h uma- nos? – Modulando la longevidad de los ratones hemos demostrado qu e los telómeros son una de esas ca usas moleculares d el envejecimie nto y que por lo tanto pueden ser clav e en muchas enferm edades asociad as al envejecimient o. – ¿Cómo actúa n esos telóme- ros? – Cuando los te lómeros son muy cortos, es to produce envejecimien to a nivel celular y causa enferme- dades. Si retras ábamos el acortamient o de los telómeros graci as a ac- tivar un enzim a lla- mada telome rasa (que es capaz d e re- parar los telom e- ros) conseguíam os retrasar, no un a, sino muchas en - fermedades a l a vez. Además, no vimos nin- María Blasco D irectora del Ce ntro Nacional de Investigaci ón Oncológica (CNIO) «No estamos p rogramados genéticament e para envejec er» María Blasco , directora del CNIO, tambié n colabora con Biodonostia. SALUD Lunes 13.0 6.16 EL DIARIO V ASCO 6 AL DÍA Francisco Real, del Centro Nacional de Investigaciones Oncológicas. J O S E L U IS P IN D A D O "En cáncer de vejiga esta- mos pasando ahora de la Prehistoria a la Historia; del Paleolítico al Neolítico". Este es el halagüeño pano- rama que Francisco Real, responsable del Grupo de Carcinogénesis Epitelial del Centro Nacional de Inves- tigaciones Oncológicas (CNIO), dibuja para el abor- daje de esta enfermedad que en España registra una de las incidencias más eleva- das del mundo: 27,9 casos por cien mil habitantes y año. Pero es que, además, desde el punto de vista bio- lógico, y también clínica- mente, es un tumor que re- presenta el paradigma de la progresión tumoral: tumo- res relativamente benignos pueden evolucionar hacia agresividad importante. "Estas variaciones son las que proporcionan intere- santes oportunidades para plantear preguntas y ofre- cer respuestas", fundamen- talmente encaminadas a evitar, en la medida de lo po- sible, cistectomías que mer- man la calidad de vida del paciente y a ofertar terapias más ajustadas a la realidad de cada tumor. DEFINIR LOS GRUPOS El primer paso para avan- zar en cáncer de vejiga es clasificar y definir tipos y subtipos tumorales. En una reunión internacional cele- brada en el CNIO y coordi- nada por Real, grupos de in- vestigación de la Universi- dad de Lund (Suecia), del MD Anderson, del Baylor College y de la Universidad de Carolina de Norte (Esta- dos Unidos), del Instituto Curie, en París, y un grupo del Cancer Gene of Atlas (TCGA), de Estados Unidos, han llegado a un acuerdo en la existencia, definición y clasificación de un grupo tumoral: el tumor Basq (ba- La clasific ió molecular a gur un 'n va er ' n cánce d v jiga El consenso mundial en la definición de diferentes grupos tumorales de vejiga abre el camino de la especialización terapéutica. Una buena noticia para un cáncer cuyo abordaje estaba estancado MADRID RAQUEL SERRANO raquelserrano@unidadeditorial.es sal squamous like). Llama- dos también escamosos, los investigadores han consen- suado, además de su nom- bre, cuáles serían los mar- cadores moleculares princi- pales que le definirían. Otro acuerdo alcanzado es la existencia de un segun- do grupo: el de diferencia- ción urotelial, aunque aún se trabaja en cuál es la me- jor forma de definirlo, por lo que no se ha estudiado a fondo, aunque la previsión para los próximos dos años es definir mejor este segun- do grupo e incluso clasificar más subgrupos que los científicos saben que exis- ten. "El Basq es agresivo. Hay muchos datos que su- gieren que, probablemen- te, es más agresivo que el de diferenciación urotelial. Re- presenta un 30 por ciento de los tumores de vejiga e in- cluye a un subgrupo que los patólogos habían recono- cido previamente y que eran tumores mixtos con diferen- ciación urotelial y escamo- sa". En este momento, varios grupos de investigación mundial trabajan para in- tentar determinar si los pa- cientes de este subgrupo responden mejor a ciertos tratamientos. TERAPIAS ADECUADAS Según Real, uno de los va- riados obstáculos a los que tradicionalmente se ha en- frentado el abordaje de esta enfermedad era la inexis- tencia de nuevos fármacos activos desde hace casi treinta años. Pero, una vez más, el Neo- lítico irrumpe, ya que en los últimos tres años se han producido significativos avances que repercutirán en los tratamientos y en el de- sarrollo de moléculas ade- cuadas a cada subgrupo, ámbito al que también se están incorporando los check points inmunes en esquemas terapéuticos. En primer lugar, se están utilizando estrategias gené- ticas para la mejor clasifica- ción molecular tumoral. "Sabemos que el cáncer de vejiga tiene ciertas similitu- des con el de mama, en tan- to que existe un subtipo de tumores vesicales denomi- nados basales, similares al basal de mama. Otros, los de diferenciación urotelial, son parecidos a los luminales de mama". ¿PRESERVAR LA VEJIGA? El cáncer de vejiga avanza- do es un punto y aparte en el que el planteamiento y objetivo prioritario es si se puede preservar la vejiga, teniendo en cuenta la mer- ma en la calidad de vida que origina la cistectomía. Para Real, en enfermedad avanzada, la radioterapia es una área donde el progre- so es más lento, pero nota- ble. "Se sugiere que los tu- mores en los que existen al- teraciones en genes repara- dores del ADN podrían ser más sensibles a la acción de la radioterapia". Por tanto, la irradiación podría ser es- pecialmente útil en casos seleccionados, donde po- dría ser incluso más eficaz que cirugía o quimiotera- pia. "Su papel debe evaluar- se y analizarse con rigor". El tumor de mama abre camino al de vejiga Identificadas ciertas similitudes entre cáncer de vejiga y mama, la hipótesis lógica es plantear si algunos de los innovadores fármacos que se emplean en mama podrían trasladarse a vejiga o incluso compartir dianas moleculares. "Es posible", afirma Real, quien explica que desde hace cuatro años se sabe que uno de los grupos de genes que con mayor frecuencia están alterados en vejiga son los que implican a la reparación del ADN, como ocurre con BRCA1 y BRCA2, con peso específico demostrado en tumores de mama. "La alteración genética responsable del tumor es más importante que el sitio donde aparece. Así, lo que se está aprendiendo en mama u ovario con los inhibidores del PARP, por ejemplo, es un objetivo para vejiga, con el inicio de estudios clínicos en pacientes con mutaciones en genes de reparación del ADN". Biológica y clínicamente, el cáncer de vejiga representa el paradigma de la progresión tumoral, hecho interesante para plantear preguntas y ofrecer respuestas" “ Grupos mundiales especializados en cáncer de vejiga hemos acordado la existencia del tipo Basq. Un segundo grupo, el de diferenciación urotelial, precisa mejor definición" “ 11/07/2016 Tirada: Difusión: Audiencia: 39.316 39.315 117.945 Categoría: Edición: Página: Rev Salud N cion l 20 AREA (cm2): 660,6 OCUPACIÓN: 61,8% V.PUB.: 2.369 NOTICIAS DE CNIO||SECTOR 28 | SOCIEDAD | Viernes, 29 de julio del 2016 | La Voz de Galicia El Tribunal Constitucional (T C) rechazó el recurso de amp aro presentado por la Asociac ión de Afectados por la Talidom ida (Avite) contra la sentencia del Tribunal Supremo para que pu- dieran ser indemnizados po r la farmacéutica que comercial izó en España este fármaco, indi ca- do en los años 60 para las n áu- seas en el embarazo hasta que se descubrió que provocaba malf or- maciones en el feto. La asociación confirmó la de - cisión del TC después de que el Supremo decidiera en septie m- bre del 2015 respaldar la sent en- cia adoptada por la Audien cia Provincial de Madrid un año an- tes, en el 2014, cuando absolv ió a la compañía alemana Grünent hal y anuló las compensaciones e co- nómicas que había fijado un j uz- gado de primera instancia. A pesar de que Avite plante ó entonces recurrir ante el Tri bu- nal Europeo de Derechos Hum a- nos de Estrasburgo, ayer el vi ce- presidente de la asociación, Ra- fael Basterrechea, declaró que es- to era «el final» y denunció q ue España es el único país en el q ue un culpable demostrado logra sa- lir impune. El recurso de amparo que Avit e presentó ante el Constitucio nal basado en que los informes q ue relacionaban el daño y el medi ca- mento estaban fechados de m a- nera errónea fue rechazado p or considerar que el de amparo no es un recurso de apelación n i el TC una tercera instancia re vi- sora, por lo que no se revalo ra- rá la prueba. Los afectados por la talidomida t ras el rechazo del Constitucional: «Es e l final» MADRID / AGENCIAS «España es el único país del mundo en el que el culpable demostrado sale libre de toda causa» Rafael Basterrechea Vicepresidente de Avite Ryanair anunció que desde e l 1 de septiembre será obligator io para aquellos pasajeros adulto s que acompañen a menores d e 12 años adquirir un asiento r e- servado, pero que ese asien to será gratuito, según dijo. Ry a- nair informa que ha decidid o poner en marcha esta iniciat i- va tras observar que había mu - chos adultos que viajaban co n niños pequeños pero no sele c- cionaban asientos reservado s, por lo que el sistema no siem - pre los sentaba juntos, ya qu e asigna asientos al azar segú n disponibilidad. Esta situación ocasionaba al- gunos problemas a la tripul a- ción durante el embarque, al intentar sentar juntos a todo s los pasajeros que volaban co n niños pequeños. La tripulació n de Ryanair no puede mover a aquellos clientes que ya ha n pagado por reservar su asien - to para sentar a otros cliente s que viajan con niños pequeño s que no han optado por reserva r. El director de márketing de Ryanair, Kenny Jacobs, dijo qu e esta nueva medida permitir á a las familias elegir sus asien - tos en el momento de efectu ar la reserva, hacer el check-in d e su vuelo hasta 30 días antes d e la salida y volar con la certez a de que siempre se sentarán co n sus hijos, y «evitará que otro s clientes con asiento reservad o tengan que ser desplazados» . Ryanair obliga a reservar el asien to gratuito para los niños pequeño s MADRID / EUROPA PRESS Si un paciente con un mela no- ma tiene una pérdida parcial de la proteína ATG5, el especia lis- ta sabrá que el pronóstico de ese caso será malo: es más proba ble que presente resistencia al tra ta- miento y desarrolle metástasi s, y podrá ofrecer un fármaco es pe- cífico o, si no lo hay, optar po r el tratamiento más agresivo. Ha sta ahora, los médicos solo pod ían basarse en el tamaño de la les ión —dos milímetros de grosor era ya una mala señal— para sa ber cómo iba a comportarse el c án- cer: una técnica aceptable p ero menos fiable que un marcad or biológico, sobre todo si se co gía de forma precoz. Este ha sido el último hallazg o del equipo liderado por la ga lle- ga Marisol Soengas, del Cen tro Nacional de Investigaciones O n- cológicas (CNIO), tras años in- vestigando qué biomarcado res pueden determinar la graved ad de un melanoma. La informac ión se ha publicado en la revista A u- tophagy. La elección de la rev is- ta no es casual, ya que la AT G5 es, precisamente, una prote ína fundamental para la autofag ia, ese proceso de autodestrucc ión que acometen las células cuan do ya no son útiles pero que, en el caso de los tumores, funcion an de una manera desconcertan te. «Con nuestro estudio queríam os descubrir hasta qué punto la au- tofagia es importante en el me la- noma en comparación con ot ras patologías, e intentar entender en la medida de lo posible esa do- ble función pro y antitumori gé- nica», explicó Soengas. Llegar a determinar la impor - Un eq ipo del CNIO aísla el gen que d termina la gravedad del mel anoma El hallazgo puede predecir el des arrollo del tumor, ahora valorad o por su tamaño Soengas lleva años invest igando los genes que dete rminan la gravedad del me lanoma. BENITO ORDÓÑEZ tancia de la ATG5 no fue sen ci- llo: «Analizamos hasta 20 ge nes de autofagia en más de 25 tip os de cáncer utilizando bases de da- tos de casi 5.000 muestras de pa- cientes —explica Soengas en el CNIO— y encontramos alte ra- ciones en el gen ATG5 que te- nían valor pronóstico solo en el melanoma». Una vez determinado el valo r del ATG5, el equipo decidió po- ner a prueba la idea, creando ra- tones genéticamente modifi ca- dos a los que les faltaba una de las copias de ATG5, y enton ces comprobaron que efectivame nte sus tumores tenían un mal p ro- nóstico, los ratones morían. In- cluso más: «Descubrimos q ue cuando los tumores pierden u na sola copia de ATG5 también r es- ponden peor a los fármacos» . La conclusión, dice Soengas, es s en- cilla: «Estamos ante un interr up- tor que regula la autofagia y fa- vorece la metástasis, y creem os que esta información nos p er- mitirá tener una mejor capa ci- dad de predecir el pronóstic o». ¿Y para qué sirve todo este co - nocimiento? «Este trabajo tie ne importantes implicaciones tr as- lacionales de cara al diseño de fármacos», apuntó Marisol So en- gas. Es decir, que la quimio te- rapia que se aplique a quien es tengan problemas con la AT G5, por ejemplo, incluya un eleme nto que fortalezca esa proteína p ara que sintetice las copias del g en. O como detector, lo que supo ne que con un análisis de sangre del paciente se sabrá si sufre esa m o- dificación de la ATG5 y por t an- to, aunque su tumor sea peq ue- ño —y el médico tienda a pen sar que controlable—, lo norma l es que sea difícil de vencer. MEDICINA DE PRECISIÓN Eficacia a la medida Todos los hallazgos de biomar - cadores tienen el objetivo de hacer pronósticos más exacto s y fármacos más eficaces. Los médicos sabrán si es conve- niente, por ejemplo, empezar con un tratamiento muy agre- sivo y dispondrán de medica- ción eficaz para esos casos. ¿PARA QUÉ SIRVE? S. C. REDACCIÓN / LA VOZ Exposición «Fotciencia». A . R. El Muncyt exhibe las fotografías ganadoras de «Fotciencia13» Cuántos gérmenes puede es- conder la mano de un niño , qué elementos oculta un cris- tal de zeolita o cómo se obtie- ne el aceite de argán son algu- nas de las preguntas que tie- nen respuesta en la exposi- ción fotográfica Fotciencia13 , que desde ayer se exhibe en el Museo de Ciencia y Tecno- logía de A Coruña (Muncyt) . La muestra reúne cerca de 50 instantáneas de las más de 700 que se presentaron a la decimotercera edición de este concurso de fotografía cientí- fica, que está organizado por el CSIC y la Fundación Espa- ñola para la Ciencia y la Tec- nología, y que tiene como ob- jetivo acercar el mundo de la divulgación a la ciudadanía a través de imágenes que abor- dan cuestiones científicas des- de una visión artística y es- tética. Los trabajos que forman parte de la exposición, que se podrá visitar hasta el 4 de septiembre, muestran una te- mática muy dispar, aunque se han agrupado en dos grandes apartados: la naturaleza en ge- neral e instantáneas obteni- das con la utilización de un microscopio. A CORUÑA / LA VOZ 06 MEDICINA MIÉRCOLES 14 SEPTIEMBRE 2016 Los tumores son un cúmulo de células que se dividen sin control, acumulando hasta cientos de alteraciones cro- mosómicas y mutaciones en su ADN. Estas alteraciones se inician en parte por un tipo de daño en el ADN co- nocido cómo estrés replica- tivo. Para sobrevivir ante este caos, las células tumo- rales necesitan de la fun- ción de la proteína repara- dora ATR, conocida por su función como guardián del genoma, a la que se vuel- ven adictas. Después de ocho años de trabajo, el equipo de Óscar Fernández-Capetillo en el Centro Nacionalde Investi- gaciones Oncológicas (CNIO) ha descubierto que el bloqueo de esta proteína tiene efectos antitumora- les en varios modelos ani- males de cáncer, como un tipo agresivo de leucemia mieloide aguda y el sarco- ma de Ewing. Los resulta- dos se recogen en sendos trabajos publicados en Science Signaling y Onco- target. El equipo de Fernández- Capetillo partía de la base Bloquear la proteína ATR tiene efecto antitumoral Inhibir la proteína reparadora ATR tendría efectos positivos en leucemia mieloide y el sarcoma de Ewing MADRID REDACCIÓN dmredaccion@diariomedico.com Una vez obtenida esta prueba de concepto, "el si- guiente paso era centrarnos en encontrar aquellos tu- mores con mayor estrés re- plicativo, ya que creemos que estos son los que más se podrán beneficiar de esta nueva terapia", señalael in- vestigador. Según datos publicados por el propio laboratorio, los tumores con mucho es- trés replicativo tienden a presentar cantidades eleva- das de la proteína CHK1 - también involucrada en su- primir el estrés replicati- vo- para sobrevivir en estas condiciones adversas. "Esto nos sugirió que una forma de identificar los tumores con más cantidad de estrés replicativo era simplemen- te evaluar cuánto CHK1 te- nían". De este modo, después de haber analizado los nive- les de CHK1 en un amplio panel de diferentes tipos de tumores humanos, los in- vestigadores identificaron dos tipos tumorales con cantidades muy elevadas de esta proteína: el sarcoma de Ewing y varios tipos de lin- fomas y leucemias, inclu- yendo la leucemia mieloide aguda. Óscar Fernández Capetillo, director de la investigación, del CNIO. D M La inhibición farmacoló- gica y genética de la vía de señalización de RANK/RANKL conduce a una reducción significati- va de recidivas y metásta- sis del cáncer de mama en un modelo animal de ra- tón, según los resultados de un estudio dirigido por Eva González-Suárez, del Instituto de Investigación Biomédica de Bellvitge (Idibell). Este trabajo, publicado en la revista Cancer Re- search, sugiere que los inhibidores RANK, que se utilizan actualmente en los pacientes con osteo- porosis y metástasis ósea, pueden tener poten- Inhibir la vía RANK reduce la recurrencia del cáncer de mama BARCELONA REDACCIÓN cial para el tratamiento del cáncer de mama. Las células precursoras de cáncer o células madre cancerígenas (CSC) expre- san altos niveles de la proteína RANK. "De he- cho, la mortalidad en el cáncer de mama se debe principalmente a las CSC supervivientes a trata- mientos, que son respon- sables de la recurrencia del tumor y la metásta- sis", explica González- Suárez. "Otros estudios de este y otros grupos de in- vestigación habían de- mostrado que la inhibi- ción de la vía de RANK podría prevenir el cáncer de mama, pero hasta aho- ra nadie había demostra- do el potencial de los inhi- bidores de la vía de RANK en el tratamiento de la en- fermedad." En su último trabajo, los investigadores de- muestran que la inhibi- ción de la vía de señaliza- ción RANK/RANKL po- dría usarse potencial- mente como nueva tera- pia en cáncer de mama; en palabras de la inves- tigadora, "esta inhibición no reduce el crecimiento del tumor, pero promueve su diferenciación, lo que reduce la población de CSC y dificulta la metás- tasis, mejorando el pro- nóstico." Cuando las condiciones son normales, el par de proteínas del meta- bolismo óseo Un equipo del Hospital John Radcliffe, en Oxford (Reino Unido), ha realizado con éxito una cirugía robótica pionera en el interior del ojo, según informa The Guardian. El cirujano Robert MacLaren y sus colaboradores han intervenido con un sistema robótico a un paciente de 70 años con una membrana epirretinia- na que le afectaba a la visión. La mem- brana tenía un espesor de entorno a una centésima de milímetro. Los especialistas aseveran que se tra- ta de la primera vez que se consigue un dispositivo que alcance la preci- sión tridimensional requerida para operar dentro del ojo humano. Tras la intervención, el paciente ha declarado que ha experimentado una mejora de la visión. Un robot opera con éxito la membrana epirretiniana Un equipo de científicos británicos ha demostrado por primera vez que se puede obtener un embrión sin utilizar gametos femeninos. Los resultados del grupo de Anthony Perry, de la Universi- dad de Bath, revolucionan los conoci- mientos asimilados en los dos últimos siglos. “Nuestro trabajo desafía el dog- ma de que sólo un óvulo fertilizado con un espermatozoide puede dar lugar al nacimiento de un mamífero viable”. El procedimiento que se describe en Nature Communications se basa en la reprogramación mitótica durante la fecundación, y se resume en la inyec- ción de esperma de ratón en embriones inactivos modificados (partenotes ha- ploides). De este modo, se ha consegui- do generar crías de ratón viables con una tasa de éxito superior al Generan embriones viables de ratón sin utilizar óvulos de que si los tumores sufren de cantidades elevadas de estrés replicativo, podrían ser especialmente sensibles al tratamiento con fármacos que inhibiesen a ATR, ya que esta proteína se encar- ga de reducir este tipo de es- trés. Además, como las célu- las sanas prácticamente no sufren de este tipo de estrés, el efecto en estas células se- ría mucho menor. "Si elimi- nas a un bombero, que sería ATR, en un pueblo donde no hay fuego, las células sanas, no pasa nada, pero si lo ha- ces en un pueblo donde hay fuego, es decir células tumo- rales con mayor daño en el ADN que las sanas,éste se extiende y destruye el pue- blo". ATACAR AL GUARDIÁN En 2011 el equipo de Fer- nández-Capetillo demostra- ba que no iba mal encami- nado. En dos trabajos inde- pendientes publicados en Nature Structural & Mole- cular Biology reportaba por primera vez que el bloqueo de ATR era especialmente tóxico en células tumorales, conclusión a la que llegaron utilizando cultivos celula- res y modelos genéticos de ratón. El cerebro de millones de personas con Alzheimer se va vaciando de neuro- nas lentamente de forma ineludible. Sin embargo, qué provoca la muerte de las neuronas es aún una incógnita. Diversos estu- dios proponen que la in- teracción de la proteína beta amiloide con la membrana de las neuro- nas provoca neurotoxici- dad. Un estudio llevado a cabo por especialistas del Estrategia obtendría agregados beta-amiloide del Alzheimer MADRID REDACCIÓN Instituto de Investigación Biomédica (IRB Barcelo- na) liderado per la inves- tigadora asociada, Natàlia Carulla, en cola- boración con científicos de la Universidad de Lo- vaina, en Bélgica, de Gro- ninga, en Holanda, y de los Servicios Científico- Técnicos de la Universi- dad de Barcelona, presen- ta por primera vez una es- trategia para obtener agregados de beta amiloi- de asociados a membrana que adoptan una estruc- tura específica de barril. Este tipo de estructu- ra –que también tienen otras proteínas de la na- turaleza–, tiene la capaci- dad de formar poros en las membranas de las cé- lulas. El descubrimiento, trasladado al contexto de Alzheimer, sugiere que el agregado también podría perforar las membranas de las neuronas, alterar- les el equilibrio celular e inducir su muerte. La descripción de estos agregados con forma de barril, publicada en Proceedings of the Im pre so po r B en ard o P os ad a V arg as. Pr op ied ad de U nid ad E dit ori al. Pr oh ibi da su re pro du cc ión . 3046068 10 11 12 16 17 9 S e acerca el momento de vivir sin prisas, de paladear la sana holgaza-nería que permiten las vacaciones. Quizá el camino hacia la felicidad la salud pase por dar valor a las cos s más simples, las que nos permiten abandonar la rutina de las pris s y reparar cuerpo y alma. Desde disfrutar de una siesta a mirar el horizonte hipnótico del mar o leer un libro tumbado sobre la hierba. El verano es tiempo de cuidarse, de ensay r nuevos háb tos saludables sin esperar a septiembre. No le estamos enviando al gimnasio o proponiendo que deje d fumar. Si lo hace, le aplaudimos. Desde estas líneas le invitamos a alcanzar tres metas más sencillas con las que además disfrutará. Coma (bien), duerma (ocho horas diarias) y medite para saborear cada segundo que viva. ¿Suena fácil? Pues probablemente lleve once meses sin cum lirlo. ¿O usted no se ha acostado algún día contestando centenares de e-mails?, ¿o despertado pensando e un taza de café tras hilvanar apenas seis horas de sueño?, ¿o sobrevive muchos días con comida precocinad ? Presumir de dormir poco y apurar la jor ad con un sandwich frente al ordenador ya no es sinónim de eficiencia y productividad. Hay empresas en Estados Unidos que empiezan a primar a sus trabaj d res si demuestran haber dormido más de siete horas. Y sería tedioso enumerar los estudios científicos que relacionan el equilibrio de nuestro organis-mo con un sueño de calidad, verdadera-mente reparador. En el último número «PLOS Medicine», un estudio de Harvard anima a cocinar en casa para luchar contra la diabetes tipo 2, una de las grandes epidemias del siglo XXI. Los que toman comida casera mantienen mejor su peso y alejan la enfermedad, defiende esta investigación. En vacaciones no hay excusa, empiece por ir al mercado, elegir productos frescos sin procesar y disfrute cada momento, como manda el mindfulness. Solo desconecte y disfrute. NURIA RAMÍREZ DE CASTRO COMER, MEDITAR, DORMIR 16 ALEJANDRO SANZ VISITA EL CNIO La Fundación Juegaterapia ha firmado un acuerdo de colaboración con la iniciativa Amigos del CNIO, del Centro Nacional de Investigacio-nes Oncológicas (CNIO), para financiar una beca de 100.000 euros que desarrollará durante dos años proyectos de investigación de excelencia en tumores pediátricos. Alejandro Sanz acompañó a la Fundación como Embajador de Honor. El cantante diseñó hace un año el pañuelo de un Baby Pelón, los muñecos que no tienen pelo y llevan un pañuelo en la cabeza en homena-je a todos los niños enfermos de cáncer. Todos los beneficios de la venta de los Baby Pelones se destinan, además de a esta nueva iniciativa, «Juegaterapia Investiga-ción», a lograr mediante el juego que los niños con cáncer pasen sus ciclos de quimio de la mejor manera posible («la quimio jugando se pasa volando»). En la imagen, el cantante, con María Blasco (centro), directora del CNIO, y Mónica Esteban, presi-denta de Juegaterapia. 20 24 Nutrición Diana Cabrera nos propone tres deliciosas y sencillas cremas frías Fotoprotección Las claves para elegir el protector más adecuado y disfrutar del sol sin riesgos CNIO Investigación Se cumplen 20 años del nacimiento de la oveja Dolly, el primer mamífero clonado SALUD 3 ABC SÁBADO, 16 DE JULIO DE 2016abc.es/salud 13 14 GM 18al24dejuliode2016 Especializada 23 IL-17A es clave en el desarrollo de la esteatohepatitis no alcohólica (NASH) El Centro de Investigación Biomédica en Red (Ciber), laUnidaddeRaquis delHos- pital laFe,el InstitutodeInvestigaciónSa- nitaria La Fe y la Universitat deValència hanpatentadounnuevokitparadiagnos- ticar la Escoliosis Idiopática del Adoles- cente (AIS),quepermiteademásestable- cer un pronóstico de la evolución de esta enfermedad. Los investigadores han en- contradomarcadores epigenéticos espe- cíficos,enparticularmicroRNAs,quepuede contribuiramejorarlacaracterizaciónde los pacientes.Hastaahora,losprincipalesestudiosde investigaciónsehancentradoenfactores genéticos y mecánicos implicados en el desarrolloyprogresióndelaenfermedad. Sinembargo,reciente ntesehadescu- bierto la participación de factores epige- néticos, que explican cómo los factores ambientales producen cambios en nues- tro fenotipo sin alterar la secuencia de ADNde la célula o tejido.En la actualidad, existe un test pronós- ticodelaEscoliosisIdiopáticabasadoenla identificacióndemutacionesgenéticas,que ayuda a establecer la probabilidad desa- rrolloalargoplazolaenfermedad.Noobs- tante,este test no permite hacer unanáli- sisdelaevolucióndeladeformidad,yaque se basa en el análisis genético puntual del pacienteyelADNdelaspersonasnosemo- dificaconel tiempo.Además,noesfactible realizar este test en la práctica clínica ha- bitual,yaquelasmuestrashandesertrans- portadasparasuprocesamientoalabora- torios especializados fueradel país. Losinvestigadoreshandescubiertouna firma compuesta por cuatro miRNAs ca- pazdediferenciarpacientesconescoliosis de controles sanos con una sensibilidad y especificidad estadísticamente significa- tiva. El método de diagnóstico/pronóstico patentadosebasaenbiomarcadoresdiná- micos en lugar de biomarcadores estáti- cos,loscualessoloofrecenunaimagenes- táticade la enfermedad.Lapatentedescribeunmétododediag- nósticobasadoenlaidentificacióndemiR- NAs circulantes en sangre como biomar- cadores en pacientes conAIS y controles sanos.“Nuestros resultados demuestran quemiRNAscirculantesenelplasmapue- denservircomobiomarcadoresparalaAIS, loqueproporcionaunnuevométodopara diagnosticar y pronosticar esta patología, evitando irradiación repetitiva de rayos x paramonitorizar laprogresiónde laesco- liosis”, destacaTeresa Bas, jefa de la Uni- daddeColumnadelHospitalUniversitario yPolitécnicoLaFe. Miembros del equipo de investigación del Centro de Investigación Biomédica enRed (CIBER) en la Universitat de València (UV) lid rado por Federico Pallardó que ha participado en esta investigación. El Ciber, La Fe y la UV patentan un nuevo kit para el diagnóstico y seguimiento de laAIS GM Madrid Elbloqueode laaccióndeestamoléculapodríaprevenir laaparicióndeestapatologíaenpacientesderiesgo Laesteatohepatitisnoalcohólica (NASH) es una enfermedad grave del hígado, antesala del carcinoma hepatocelular (CHC),y que actualmente no tiene trata- miento. Un estudio desarrollado en el Centro Nacional de Investigaciones Oncológicas (CNIO) señala que la moléculaproinflamatoria IL-17Apodría ser el protagonista en el des rrollo de estapatología,yapuntaaquesubloqueo o la inhibiciónde lascélulasque lasecre- tan con fárm os como la digoxina prevendría su aparición en p cientes con riesgo de desarrollar este tipo de cáncer. Nabil Djouder, líder del estudio, explica a GM que IL-17A puede servir como biomarcador no invasivo del proceso inflamatorio.“Podría ayudar a identificar a los pacientes en riesgo de desarrollar NASH y CHC”, subraya, añadiendo que este hallazgo propor- ciona “nuevas estrategias preventivas y terapéuticas”.Ladigoxinayaestádispo- nible y se utiliza ocasionalmente en el tratamiento de diversas enfermedades del corazón, como fibrilación auricular (FA), o insuficiencia cardiaca (IC). Trabajandocondiferentesmodelosde ratón,losautoresmuestranenel trabajo publicadoenCancerCell cómoel exceso denutrientesestimula laexpresiónenel hígado de un oncogen llamado URI.URI —que también está sobreexpresado en las infeccionesporvirusde lahepatitisB y C— provoca daño en el ADN de los hepatocitosyestodesencadena inflama- ción sistémica y un diálogo entre el hígado y el tejido graso que culmina con la aparición de NASH. HepatitisydiabetesCuando se produce el daño en elADNde los hepatocitos, las células inmunes se infiltran en el hígado, especialmente los linfocitosTh17. IL-17A provoca la infil- tración de neutrófilos en el tejido graso que causa resistencia a la insulina y la liberación de ácidos grasos que se acumulan en el hígado o, lo que es lo mismo,NASH. “Hasta el momento,no está claro si la hepatosteatosis era una causa o una consecuencia de disfuncionesmetabóli- cas,aunquenuestroshallazgossugieren que los pacientes con diabetes y, por tanto, con resistencia a la insulina pueden acumular grasa en el hígado a través de comunicados de ácidos grasos del tejido adiposo blanco”, expone Djouder. “Esta acumulación de grasa puedepropiciar el desarrollo deNASHy deCHC.Esmás, los pacientes condiabe- tes corren el riesgo de desarrollar NASH”,agrega.En este sentido, el trabajo concluye destacando que el tratamiento de la hepatitis C representa un reto socioeco- nómico en nuestra sociedad, por lo que el bloqueo de la acción de IL-17A con digoxina proporcionaría una forma de profilaxis efectiva y barata para los pacientes con hepatitis B o C,que tienen unelevadoriesgodedesarrollarNASHy carcinoma hepatocelular.“Tratar la obesidad puede ayudar a tratar muchas otras disfunciones metabólicasy,aunque laacumulaciónde grasa no pueda explicar el resultado negativo deNASH,la inflamación deter- mina el pronóstico a largo plazo de la enfermedad”, afirma. Por tanto, el hallazgo de nuevos mediadores de la inflamación y su bloqueo pueden ser futuras terapias estratégicas. A la izquierda, una imagenhistológica de unhígado de ratón sano y, a la derecha, de unhígado de ratón con esteatohepatitis no alcohólica.FOTO: CNIO J.V. Madrid 15 UN CANDIDATO ESPAÑOL AL N NURIA RAMÍREZ DE CASTRO MADRID E l próximo 3 de octubre la Academia Nobel anuncia-rá el premio de mayor prestigio en investigación médica. En las quinielas de los posibles candida-tos d stacan los descubridores de una herramienta de nombre impronuncia-ble –CRISPR (léase crisper)– que está revolucionando los laboratorios de todo el mundo. Esta técnica permite cortar y pegar genes a voluntad; en de-finitiva, cambiar el destino escrito en el ADN y editar cualquier forma de vida. Lo hace, además, d una forma tan sencilla, económica y eficaz que «asusta» a sus propios creadores. Su potencial es asombroso. Podría mejo-rar cultivos y hasta resu-citar especies extintas. Pero sobre todo tiene un asombroso potencial te-rapéutico con enfermeda-des tan difíciles de tratar como el cáncer o miles de enfermedades heredita-rias. Si el «corta-pega» ge-nético no recibe el reco-nocimiento del Nobel este año, lo hará con toda se-guridad en los próximos cinco años, auguran los gurús de la ciencia. Gane o no, detrás de esta asom-brosa herramienta hay un nombre español, el del bió-logo de la Universidad de Alicante Juan Francisco Martínez Mojic . Descubrió que las bacterias tienen su propio sistema in-mune y con ese hallazgo abrió la puer-ta a est revolucionaria máquina de manipulación g é ica. Su historia es la de una investigación con final feliz, pero en la que durante años no se vis-lumbraba una utilidad. «Seguí inves-tigando por cabezonería, casi fue una cuestión de fe», explica Mojica a ABC durante su visita a Madrid para parti-cipar en un ciclo de conferencias del Centro Nacional de Investigaciones Oncológicas (CNIO), patrocinado por la Fundación Banc Sabadell. —¿Cómo fue su momento eureka? —Todo empezó en las salinas de San- Martínez Mojica, el cerebro alicantino del «corta-pega» genético· Es el «padre» de una técnica revolucionaria que permite cambiar el destino es rito en los genes ta Pola, mientras trabajaba en mi te-sis doctoral. Estaba estudiando cómo unos microorganismos de las salinas eran capaces de adaptarse a condicio-nes de vida extrema. Empecé a estu-diar su genoma y encontramos unas repeticiones del genoma que se encon-traban espaciadas a intervalos regu-lares. Es decir, entre una repetición y la siguiente había una secuencia dis-tinta de un tamaño constante. Enton-ces no nos imaginábamos la rep rcu-sión que podía tener. Pero seguimos trabajand . Al principio, todos los ex-perimentos fueron un fracaso. Fue ab-solutamente desesperante. Yo seguí investigando por cabezonería, algo me decía en mi interior que aquellas se-cuencias eran importan-tes. Me marqué averiguar qué función estaban cum-pliendo como objetivo de mi carrera científica. Frus-trado porque no avanzaba, decidí revisar los genomas completos de bacterias se-cuenciadas. Así describí una nueva familia de repe-ticiones que llamé CRISPR. Pero aún no sabíamos para qué servían estas repeti-ciones. El momento eureka llegó en 2003 cuando vi-mos claramente que era un sistema inmunológico de las bacterias. —Se ha dicho que duran-te esos años de búsqueda no contó con el suficien-te apoyo del Gobierno. ¿No supieron ver el poten-cial de su trabajo? —Si yo hubiera sido un evaluador de proyectos científicos en aquel momen-to no me lo habría dado a mí mismo (risas). Creo que es una respuesta lo suficientemente clara a su pregunta. Quienes evalúan deben dar dinero a grupos competitivos, que producen y vean posibilidades de éxito. Yo pasé unos años sin publicar y entiendo que no me dieran financiación. En ese mo-mento era imprevisible lo que iba a sa-lir de allí y llegar a la locura que esta-mos viviendo ahora. —Y ahora ¿se siente apoyado? —Sí, m y apoyado. Ha habido un cam-bio en los últimos años, incluso antes de que se me reconociera como el «pa- CRISPR podía servir para editar geno-mas. Si se hubiera reconocido el he-cho de quien dio pie al desarrollo de esa tecnología, entonces sí, mi nom-bre debería haber estado allí. —Pero usted mismo acaba de decir «yo lo parí» —Sí, pero todo depende de lo que que-ramos mirar hacia atrás. —Meses más tarde llegó el Jaime I. ¿Siente que fue un premio de conso-lación? —Personas relevantes del mundo de la ciencia me han dicho que no enten-dían cómo no me habían concedido el Princesa de Asturias, sobre todo cuan-do las premiadas habían reconocido que yo era el padre de la técnica. Yo es-toy muy satisfecho con la concesión del Jaime I y prefiero creer que me lo han dado porque me lo merezco. —¿Cómo se puede explicar la expan-sión de esta técnica? —Ha sido bestial. Antes había otras herramientas para editar genomas. drino» de estas herramientas. —Su trabajo fue el punto de partida de una revolución científica que es-tán aprovechando otros científicos. ¿Esto es frustrante o es, sin embar-go, la mejor recompensa? —Yo estoy absolutamente satisfecho por haber contribuido. Creía en ello y ha resultado ser algo excepcional. —El último Princesa de Asturias de Investigación premió a dos científi-cas por esta técnica, Doudna y Char-pentier. ¿No echó en falta su nombre? —Me alegré una barbaridad de que se lo dieran. Recuerdo que el día de la concesión del premio entró uno de mis colaboradores en mi despacho y me dijo: «¡Francis, han dado el Princesa de Asturias a CRISPR!», y así es como yo lo vi. Por primera vez había un re-conocimiento al sistema que, por de-cirlo de alguna manera, parí yo. El pre-mio reconoce el desarrollo de una he-rramienta y ellas fueron las primeras personas que dijeron que el sistema CNIO Legislación «Cualquiera puede utilizar esta tecnología tan potente. Necesitamos regularla» Ensayos «Da miedo que se esté probando ya. Los efectos sencundarios no tienen marcha atrás» Ã El investigador, en el Centro de Investigación Oncológica 44 SOCIEDAD abc.es/conocer SÁBADO, 24 DE SEPTIEMBRE DE 2016 ABC 18 SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 183 19 QUO, September 27, 2016 20 El Pais Semanal, October 9, 2016 21 El Mundo, October 15, 2016 22 La Razón, October 19, 2016 23 Orbita Laika, October 19, 2016 24 En Intermedio, La Sexta, November 2, 2016 25 La Razón, November 18, 2016 26 El Mundo ( front page ), November 25, 2016 57EL PAÍS SEMANAL / 9.10. 2016 María Blasco “La ciencia no debe renunciar a la rentabilidad” La investigadora y directora del CNIO asegura que la apuesta clave de la ciencia es desentrañar el envejecimiento para manipular genéticamente ese proceso. por José Luis Barbería F O T O G R A F Í A Gorka Lejarcegi 2089 DOC maria blasco.indd 57 28/09/2016 16:45:32 EL MUNDO. SÁBADO 15 DE OCTUBRE DE 2016 28 EM2 i CIENCIA MIGUEL G. CORRAL MADRID Francisco J. Ayala (Madrid, 1934) es una referencia científica internacio-nal, una de las máximas figuras mundiales en Genética y Evolución. Ostenta el título exclusivo de Uni-versity professor, el más elevado de la Universidad de California, Irvine (EEUU), a pesar de que tres de sus colegas son Premio Nobel. Su agra-decimiento a esta institución le ha impulsado a donar casi 12 millones de dólares, entre premios recibidos y dinero propio obtenido con sus actividades como productor de uva destinada a la producción de vino en California. Acaba de visitar Ma-drid para impartir una conferencia en el ciclo Distinguished Seminars del Centro Nacional de Investigacio-nes Científicas (CNIO), patrocinado Fundación Banco Sabadell. Pregunta.– ¿Está el hombre mo-dificando la evolución con los avan-ces en Biología Sintética? Respuesta.– No, igual que si crea-mos una máquina o un objeto de ar-te o una casa no estamos jugando a ser dioses. Hacer células es mucho más difícil, más complejo, que hacer una casa. Así que es algo que va a llevar tiempo. Ya se están constru-yendo, pero son células muy sim-ples y además se utilizan partes pre-existentes. Todavía no hemos llega-do al estado en el que podamos crear realmente una célula, crear la vida, desde el principio, desde com-ponentes puramente inorgánicos. P.– Si somos capaces de manipu-lar su ADN sí podemos alterar lo que trasmitirán a su descendencia. ¿Eso no es alterar la evolución? R.– Estamos aprendiendo cada vez más cómo modificar el genoma, el ADN, de los organismos. Todavía lo hacemos bastante por tanteo y de una manera un poco burda. Aunque en los últimos tres años hemos des-cubierto estos nuevos métodos que se llaman CRISPR, que abren la po-sibilidad de enfocarse en un gen único. Pero todavía son cambios que se modifican en las células so-máticas, no en las reproductivas. Esto no sabemos cómo hacerlo, no podemos modificar el semen o los óvulos. Quiere decir que estas mo-dificaciones que hacemos se que-dan en esa generación, no se tras-miten a la generación siguiente. P.– El español Francis Mojica es uno de los pioneros de esta técnica. ¿Cree que podría obtener el Nobel? R.– Doudna y Carpentier son las que lo obtendrán con seguridad. Y hay algunos otros científicos, no só-lo Mojica, también un investigador chino del MIT, que podrían ser mere-cedores. Así que sí, lo podría obtener. Espero que Mojica sea el tercero. P.– ¿La medicina está yendo en contra de la Selección Natural? R.– Naturalmente que sí. Hay agoreros que dicen que cuanto más avance la medicina y más enferme-dades curemos, más enfermedades habrá que curar en la generación próxima. Pero en una enfermedad recesiva con una incidencia de una entre 10.000 personas, dentro de 100 generaciones va a ser de cuatro en 10.000. De aquí a entonces ha-brán avanzado tanto los fármacos y la medicina que no habrá que preo-cuparse. La medicina tiene menos de 1.000 años y la genética diría que menos de 100 y los avances han si-do muy grandes, imagine lo que se puede hacer en 30 siglos. P.– ¿Ni siquiera el sumatorio de una y otra enfermedad le preocupa? R.– Es cierto que este proceso ocu-rre, que hay que curar más enferme- dades en las generaciones siguien-tes. Pero desde el punto de vista de la población humana el efecto es tan pequeño, que siempre que podamos curar hay que hacerlo, tenemos la responsabilidad de mejorar la vida humana todo lo que se pueda. P.– ¿Qué le falta a la política cientí-fica española? R.– La política científica españo-la sufre de que simplemente el país no invierte suficientemente en cien-cia. EEUU invierte el 3% del PIB en investigación científica y España el 1,2%. Cuando murió Franco era el 0,4% y en los 80 llegó hasta el 1%, pero nunca se ha superado el 1,3%. Los Gobiernos no entienden los beneficios de la investigación cien-tífica. El 50% de todos los avances económicos en EEUU desde la Se-gunda Guerra Mundial se deben a descubrimientos científicos. Es de-cir, el Gobierno invierte el 3% y ob-tiene el 50% de sus avances econó-micos de esa inversión. P.– ¿Falta también una cultura del mecenazgo privado? R.– Sí, el problema es la falta de tradición en España de ser generoso y de contribuir a la ciencia y a la be-neficencia. La muchacha de servicio que trabaja para nosotros en casa da más dinero a la Iglesia y a obras be-néficas que lo que dan mis herma-nos en Madrid, y ellos tienen mucho más dinero, pero dan una pequeñez. P.– Hay colegas que dicen que reli-gión y ciencia no son compatibles… R.– La metáfora que yo uso es que la ciencia y la religión son dos venta-nas diferentes para ver el mismo mundo, pero lo que se ve es diferen-te. La ciencia se ocupa de la evolu-ción biológica, la astronomía… La religión se ocupa de los valores mo-rales. Son cosas diferentes. P.– Pero en EEUU hay quienes se empeñan en mezclarlo, me refiero a los creacionistas. ¿No cree? R.– La ciencia no sólo es compa-tible con la religión, sino que es mucho más compatible que sus creencias, que son implícitamente blasfemias. Porque si uno cree que Dios ha diseñado el cuerpo huma-no tal como es, entonces cómo ex-plicar todos los defectos que tene-mos. Incluyendo el hecho de que el 20% de todas las concepciones hu-manas terminan en aborto espon-táneo durante el embarazo. Eso son 20 millones de abortos al año debido a que el sistema reproduc-tivo humano está muy mal diseña-do. Si le van a echar la culpa a Dios por ser el abortista mayor del mun-do, eso es una barbaridad. P.– ¿Con los negacionistas del cambio climático ocurre algo pare-cido en EEUU? R.– Un señor que aspira a la Casa Blanca dice que el cambio climático es algo que han inventado los chi-nos para perjudicar a la economía americana. Y es candidato a ser presidente de los Estados Unidos. P.– Pero da la sensación de que el negacionismo climático tiene me-nos presencia, ¿todavía existe el riesgo de que se dé marcha atrás? R.– No creo que dé marcha atrás. Y primero, no creo que lo elijan presidente. Además, no quiere de-cir que lo que dice ahora que hará sea lo que vaya a hacer de verdad. Aunque en EEUU todavía hay mu-chas personas que no creen que la contribución humana sea impor-tante en el calentamiento global. P.– Suena a acto de fe… R.– Es una actitud muy parecida, simplemente negar la evidencia porque no es consistente con mis propias convicciones, porque tengo inversiones en carbón o porque me gusta usar gasolina para el coche... P.– ¿Y si Donald Trump llegase a ser presidente de EEUU? R.– Las consecuencias son im-predecibles, como lo es él. Pero si fuera a actuar con arreglo a lo que dice, sería una catástrofe. «El 50% de todos los avances económicos de EEUU se deben a hallazgos científicos» «En España, no se invierte suficiente en ciencia. Se dedica sólo el 1,2% del PIB» ALBERTO DI LOLLI «Los Gobiernos no entienden los beneficios de la investigación» Catedrático de la Universidad de California, Irvine (EEUU). Es una eminencia mundial en el campo de la Biología Evolutiva y tras donar cerca de 12 millones de dólares a la universidad que le ha acogido también es uno de los mayores filántropos científicos FRANCISCO J. AYALA «Ciencia y religión son dos ventanas diferentes para mirar al mismo mundo» «Si Trump llegase a presidente y hace lo que dice que hará, sería una catástrofe» 19 20 21 26 Navidades , aunque p arece que le adelant arán una de las sesiones p ara que no le coincid a con Noch ebuena. «A sí, a lo me jor puedo comer alg unas gam bas, aunqu e lo mismo acabo tom ando un c aldito», dic e. Tiene pensado h acerse un tatuaje qu e ponga «never giv e up», «nu nca te rind as», una fr ase «LA PRIME RA SESIÓN, BI EN; EN LA SEGU NDA OCASIÓN T UVE VÓMITOS» , RELATA «que me s ervirá para toda la vid a». Acabad as las 16 sesio nes de qui mio, le qu edan 30 d e radio. Las va a comp aginar con su trabajo de teleasisten cia en Aten zia, donde ayuda a personas c on discapa cidad. «Si he podido con 16 de quimio, ¿c ómo no vo y a poder c on 30 de radi o? Hay qu e echarle h umor a es to». La misión d el investigad or Javier Benítez, desd e hace dos dé cadas, es identifi car to dos esos gene s respon- sables de qu e las mujeres desarro- llen cáncer d e mama, al m argen de los factores a mbientales. E s el direc- tor del Progr ama de Gen ética del Cáncer Hum ano del Cent ro Nacio- nal de Invest igaciones On cológicas (CNIO) y des de que en 199 5 se iden- tifi caran los p rimeros gene s respon- sables del c áncer de hu mano, el BRCA1 y el B RCA2, ha seg uido se- cuenciando las mutacion es. En 2001 pus ieron en ma rcha la consulta de cáncer famil iar. Fue la primera de E spaña. Dos a ños más tarde pasó a fi nanciarla la Comu- nidad de Ma drid. Y es qu e una de cada ocho m ujeres tiene probabi- lidades de padecer tu mor de mama. Poco a poco han ido am- pliando el e spectro de i nvestiga- ción y, mien tras «antes e xigíamos que al meno s tres familia res hubie- ran tenido c áncer y uno de ellos fuera menor de 50, ahora también investigamo s casos aisl ados de gente joven que tienen u n tipo de tumor concr eto», explica . Y es que, gracias a sus i nvestigacion es fueron capaces de id entifi car que , entre el 60 y el 70 por ciento de las mujeres portadoras d el BRCA1 tie nen pro- babilidades de desarroll ar cáncer de mama. Bá rbara Hernán dez, que aparece en l a foto junto al doctor, B. V. CONQ UERO - Mad rid tiene e sta mutación . Pero ella ent ra dentro de las nuevas paut as, ya que le detectaron el tumor con 30 años, muy joven y previamen te no lo había desarr ollado ningu no de sus familiares. Ya ha tenido q ue pasar por tres ope raciones, ad emás del tratamiento de quimi oterapia, pero su opti mismo le ha ayudado a superarlo. «Ahora soy ‘‘coach’’ emocional y ayudo a la g ente con cáncer a sob rellevarlo», a fi rma. Mientras Bár bara se centr a en el bienestar de los paciente s, Benítez participa en un proyecto europeo gracias al que han consegu ido iden- tifi car otros t antos genes de agresi- vidad mod erada, grac ias a los avances tecn ológicos que permiten hacer una se cuenciación masiva. En una terce ra vía de inve stigación también trab ajan en gene s de bajo riesgo que af ectarían a en tre el 11 y al 12 por cien to de los tum ores. Sin embargo, au nque parezca que se ha avanzado m ucho, como explica Benítez, «si sumamos BRCA1, BRCA2, los m oderados y lo s de bajo riesgo sólo t endríamos i dentifi ca- dos el 50 por ciento de las mutacio- nes genétic as responsa bles del cáncer de m ama». Sabe que aún queda much a labor por ha cer, pero gracias a esto s avances tam bién se pueden iden tifi car mucho mejor las posibilidade s que tiene u na mujer de tener un tu mor y hacer u n criba- do más espe cífi co y meno s dañino que la mamo grafía. Aliarse con la genética y predecir e l tumor LUCHA C ONTRA LAS MUT ACIONE S Cristina B ejarano La inmunote rapia tiene u n impac- to decisivo e n detener los tumores más letales. Esta es la co nclusión del último co ngreso de la S ociedad Europea de Medicina On cológica (ESMO, en s us siglas en ingles). Y uno de los m ayores impu lsores de estos nuevo s tratamien tos que trabajan co n nuestras propias defensas es I gnacio Mele ro, inves- tigador de in munoterapi a experi- mental del cáncer de l a Clínica Universidad de Navarra , y cuyo proyecto ha recibido la fi nancia- ción de más de un millón de euros de la Aecc. «La inmun oterapia benefi cia a e ntre un 20 y u n 50 por ciento de lo s enfermos d e cáncer y poco a p oco estamo s consi- guiendo qu e su uso sea más fre- cuente», exp lica. ¿Qué es exac tamente la in muno- terapia? «Es un tratamien to por el que nuestro sistema inm unitario nos ayuda a matar las cé lulas in- fectadas». Consiguen mandar instruccione s a nuestros l infocitos gracias a cier tos fármacos para que ataquen a lo s tumores y, así, «ob- tenemos re sultados du raderos sobre la prog resión del tum or». Por el momento , los mejores resulta- dos se han obtenido en la lucha contra el me lanoma en m etástasis, que fue el p rimero en e l que se probó, y en e l linfoma de Hodgkin que, «en los estadios ref ractarios B. V. CONQ UERO - Mad rid consi gue control ar la enferm e- dad», aunq ue también se están consiguiend o buenos re sultados en cáncer d e cabeza y c uello, así como en el r enal de célul as claras. En lo que se refi ere al de mama están trabaj ando con un o de los más agresivo s, el carcinom a triple negativo y « hemos com probado que existen buenos resu ltados en un 18-25 po r ciento de l os casos que hemos analizando, aunque ahora estam os reclutand o a pa- cientes para verifi car los resulta- dos en segu nda línea d e trata- miento», aún queda much o traba- jo, «creemos que a media dos o fi - nales de 201 7 podremos tener el primer fárm aco». Y es q ue esta nueva realid ad ha alenta do a mu- chas enferm as de mama a sumar- se a los ens ayos clínico s que se están realiz ando en di ferentes hospitales e spañoles. Lo s benefi - cios de esta t erapia puede n marcar el devenir de una pacient e: puede alargar la su pervivencia meses. En lo que se refi ere a los efectos secundarios , los resultad os tam- bién son bue nos porque «el perfi l de tolerabi lidad es bu eno. Se pueden de spertar rea cciones autoinmun es e infl am atorias, pero son pr oblemas de fácil tra- tamiento». Es por todo esto por lo que ya se h an aprobado fárma- cos para ha sta nueve p atologías y más de 2. 500 pacient es se be- nefi cian de ellos. Primer fárm aco contra e l cáncer de m ama en 201 7 EL FUTU RO DE L A INMUN OTERAP IA SEÑALES D E ALERTA DE LA ENF ERMEDAD La doctora Marta Blan co, de la Asociac ión Españo la Contra el C áncer (AEC C), establece los princip ales signos de alarma:  Aparición de nódulos e n la mama o e n la zona d e la axila.  Irregularidad es en la zo na de la mam a o hundim ento del pezón . También hay que vigilar cua lquier alter ación en el cont orno de la mama.  Desarrollo d e piel de naranja en el pecho ( con una aparie ncia simila r a la de la celul itis), aunqu e a veces pue de parece r simpleme nte un ecc ema.  La secreción espontán ea a través del pezón.  ¿Es útil la aut oexplora- ción? Para la doctora , no está muy r ecomend ada y generalme nte no sirv e. Asegura q ue se lleva n a cabo biop sias y una serie de pruebas si n necesida d, si bien ante c ualquier sí ntoma siempre h ay que con sultar con el mé dico.  En cuanto a los quistes mamarios , en mujer es jóvenes, u no de cad a cuatro son benig nos.  Según una e ncuesta a 2.621 muje res realiza da por el centro S alud de la Mujer Dexeus, un a de cada cuatro cree, de fo rma errón ea, que la principa l causa de l cáncer de mama es la predisposi ción gené tica hereditaria . SOCIEDAD DÍA MUN DIAL CON TRA EL C ÁNCER D E MAMA 39 LA RAZÓ N • Miérc oles. 19 de octubre d e 2016 23 22 B. V. CONQUERO / J. V. ECHAGÜE culares de la Neoplasias Linfoides del IDIBAPS que dirige Elías Cam- po. Llevan años trabajando en las células de la sangre y, en concreto, en los linfocitos, «los defensores de nuestras células sanguíneas», apunta Campo. Él también forma parte del consorcio europeo Blue- print: «Llevamos seis años traba- jando de forma coordinada», añade. Lo que se habían dado cuenta los investigadores del Clínic es que no todos los pacientes tenían la misma esperanza de vida a pesar de padecer el mismo cáncer, un linfoma. «Unos viven muchos años, mientras que en otros la enfermedad era mucho más agre- siva». Como explican en el estudio que han publicado en «Cancer Cell», «la célula del cáncer es dife- rente cuantitativamente respecto a la célula normal. Así, ser más o menos agresivo está directamen- te relacionado con los cambios del epigenoma», sostiene Campo. De ahí que, cuando hablamos de al- teraciones en el genoma, nos re- ferimos a mutaciones, mientras que las que corresponden a la epigenética se defi nen como «cambios». «No existe ninguna evidencia de que existe alguna infl uencia ambiental para que un linfoma sea más agresivo que otro, pero sí que hemos podido determinar con este estudio que los cambios en la edad de la célula son claves». Algunas viven horas, días, meses o años. Según Iñaki Martín-Su- bero, del mismo equipo que Campo, «en nuestro estudio, el factor más importante para ex- plicar la agresividad clínica es el ta para elaborar un buen plato», explica a LA RAZÓN Enrique Carrillo, investigador postdocto- ral que ha trabajado a las órdenes de Alfonso Valencia en el Progra- ma de Biología Estructural y Biocomputación del Centro Na- cional de Investigaciones Onco- lógicas (CNIO). Junto al Instituto Nacional de Bioinformática de este mismo organismo, constitu- yen la participación española en el IHEC, el mayor estudio euro- peo del epigenoma humano. Ayer se publicaron 41 trabajos de forma simultánea, 24 de los cua- les aparecieron en varias revistas del grupo «Cell Press». La parte europea, conocida como Blue- print, se ha encargado de hacer accesibles los datos generados, así como de coordinar su análisis entre más de 40 universidades europeas. El «nivel de complejidad ha sido mayor» en el estudio del epigeno- ma. No en vano, el genoma es único para todas las células del cuerpo. Pero a nivel epigenómico, cada célula tiene una información diferente: es éste el que va a deci- dir que una célula pertenezca al páncreas, otra a un músculo... y también, que lleguemos a desa- rrollar algunas enfermedades. A nivel mundial, se han estudiado más de 2.000 epigenomas de pa- cientes que sufren males hereda- dos. Así, el estudio también han buscado las claves que explican cómo las células se alteran en función de una enfermedad como el cáncer. En esta área es en la que se ha centrado un equipo del Hospital Clinic de Barcelona, en concreto el de Mecanismos Mole- nes específi cas para cumplir con las funciones que tienen enco- mendadas. «El genoma lo cono- cemos. Son letras seguidas unas detrás de otras. Pero no sabemos cómo ordenarlas. Es como si tu- viéramos ‘‘El Quijote’’ con todas sus palabras, pero sin signos de puntuación: comas, puntos, etc. Y si no están en su contexto, no lo entendemos. En eso consiste el epigenoma: los procesos quími- cos que regulan las células. En el genoma tenemos los ingredien- tes, pero el epigenoma es la rece- Hace unos años, un grupo de científi cos dio un paso de gigan- te: descifrar el genoma, ese disco duro con diferentes programas que marca el devenir de nuestras células. Sin embargo, existe un segundo nivel, conocido como epigenoma que representa el trabajo de cada una de esas célu- las. El corazón, el hígado, el pán- creas... cada uno de estos órganos tiene un epigenoma diferente que les permite seguir las órde- SOCIEDAD El mayor estudio europeo sobre el epigenoma, con participación española, explica el comportamiento de las células y promete futuros tratamientos para enfermedades como el cáncer El gran libro de instrucciones de los genes 42 Viernes. 18 de noviembre de 2016 • LA RAZÓN SALUDE M 2 38 EL MUNDO. VIERNES 25 DE NOVIEMBRE DE 2016 CRISTINA G. LUCIO MADRID En 2006, el japonés Shinya Yama-naka descubrió que era posible dar marcha atrás en el reloj de las células y conseguir que retrocedie-ran a un estado similar al embrio-nario. La reprogramación celular ha supuesto, desde entonces, toda una revolución para la ciencia, aunque sigue siendo mucho lo que se desconoce sobre el proceso. Un equipo del Centro Nacional de Investigaciones Oncológicas (CNIO) ha contribuido esta sema-na a despejar un poco más esas incógnitas al describir uno de los mecanismos clave que explican la reprogramación. Según sus datos, publicados en la revista Science, cuando la reprogramación ocurre en un tejido, el daño tisular es un factor relevante para revertir el es-tado de las células. La técnica de Yamanaka se basa en la introducción de un cóctel de cuatro genes denominado OSKM. La combinación de estos factores hace posible que una célula adulta, como por ejemplo una neurona, vuelva a convertirse en una célula parecida a la embrionaria (o pluri-potente) y, por tanto, con capacidad para convertirse en cualquier otra célula específica del organismo. Hasta 2013, todos los intentos de reprogramación se habían he- cho en el laboratorio, en placas de cultivo. Pero entonces el Grupo de Supresión Tumoral del CNIO, li-derado por Manuel Serrano, logró reprogramar células en el interior de un organismo vivo –un ratón transgénico–, abriendo la puerta, entre otras cosas, a un mejor co-nocimiento del proceso. La continuación de su investiga-ción –en la que también ha parti-cipado la directora del CNIO, Ma- ría Blasco– les ha permitido, de hecho, descubrir que el daño tisu-lar cumple un papel muy impor-tante en la reprogramación. «Cuando activamos los factores de Yamanaka, algunas células dife-renciadas sufren un daño», explica Lluc Mosteiro, investigadora del CNIO y principal firmante del tra-bajo que publica Science. Y ese da-ño, continúa, «favorece el proceso de reprogramación de las células vecinas» que se ha iniciado con la introducción del cóctel de genes. El descubrimiento supone todo un cambio en este campo de investiga-ción ya que hasta ahora se conside-raba que el daño celular provocado por OSKM no cumplía ningún pa-pel en el proceso de reprograma-ción o que incluso contribuía a re-ducir la eficacia de la técnica. En concreto, los investigadores del CNIO han observado que las células lesionadas secretan unas señales que son capaces de favo-recer la reprogramación en las células vecinas del mismo tejido. Estas señales, entre las que des-taca una molécula, la interleuki-na-6, son mediadores clave para conseguir la reprogramación, subrayan los investigadores. Para compro-bar la relación entre el daño ce-lular y la repro-gramación, los científicos del CNIO manipula-ron el proceso y vieron que, generando un daño extra a cé-lulas pulmonares de ratones trans-génicos, «la re-programación se veía muy favore-cida» en las célu-las adyacentes. Del mismo modo, también comprobaron que el envejeci-miento –que se asocia con mayo-res niveles de senescencia celular, un tipo de alteración por el que las células dejan de dividirse aunque no mueren– también potencia la reprogramación iniciada por los factores OSKM introducidos. Pero el hallazgo de los investi-gadores del CNIO va mucho más allá del proceso de reprograma-ción celular y podría ser también útil para comprender el proceso natural de regeneración, lo que tendría importantes implicacio-nes médicas. TEJIDO LESIONADO «Nuestra hipótesis es que cuando un tejido se daña de manera fisio-lógica, es posible que la respues-ta del organismo para regenerar ese tejido pase por un proceso si-milar al de la reprogramación», explica Mosteiro. Según esa teo-ría, las células adultas darían marcha atrás en su estado para aumentar su potencial regenera-tivo y poder afrontar la curación del tejido lesionado, señala la in-vestigadora, quien subraya que precisamente este punto «es una de las líneas que se van a investi-gar ahora». El grupo del CNIO también in-tentará probar distintas combina-ciones farmacológicas para, una vez conocido el papel esencial de la interleukina-6, conseguir mejorar aún más el proceso de reprograma-ción e, incluso, intentar poner en marcha el proceso sin necesidad de usar los factores de Yamanaka. «Los genes de Yamanaka son poco eficientes induciendo repro-gramación», señala Mosteiro, por lo que encontrar nuevas estrate-gias para mejorar el proceso sería de gran ayuda para la investiga-ción. Además, si se confirma que este proceso tiene lugar durante la reparación natural de tejidos, po-dría abrirse una importante vía para la medicina regenerativa, subraya la autora del estudio. CUANDO SE UTILIZAN LOS FACTORES DE REPROGRAMACIÓN, LAS CÉLULAS SUFREN UN DAÑO QUE ES FUNDAMENTAL LAS CÉLULAS LESIONADAS EMITEN UNAS SEÑALES QUE FAVORECEN LA REPROGRAMACIÓN EN OTRAS VECINAS El investigador del Centro Nacional de Investigaciones Científicas Manuel Serrano, en una foto de archivo. NACHO ALCALÁ La investigadora y directora del CNIO, María Blasco, en su laboratorio. ALBERTO DI LOLLI BIOMEDICINA LA CLAVE ES EL ENVEJECIMIENTO O EL DAÑO EN EL TEJIDO CÉLULAS MADRE ‘CONTAGIOSAS’ Científicos españoles descubren el mecanismo por el cual las células adultas retroceden hasta un estado similar al embrionario y se reprograman para ser como las del tejido en el que se implantan 25 26 24 ANNUAL REPORT 2016 184 COMMUNICATION INVITED GUEST SPEAKERS ( Distinguished Seminar Series ) Edurne Pasaban, March 8, 2016 Diane Simeone, June 27, 2016 Francisco J. Martínez Mojica, September 16, 2016 Francisco J. Ayala, October 14, 2016 Charles Brenner, October 28, 2016 Celeste Simon, December 2, 2016 SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 185 SOCIAL EVENTS In 2016, the CNIO once again, partic- ipated in the European Researchers ’ Night ; an event funded by the EU Framework Programme. Over 200 participants visited the CNIO that opened its doors to the public as part of its commitment to scientific dissem- ination and education. The event pro- vided guests with the opportunity to meet researchers in an interactive and entertaining way. September 30, 2016. The first distinction received by the ‘CNIO Friends’ initiative came from social media. Indeed, its Facebook page rose to first place in its category in the Internet Day Awards, sponsored by Google, Fénix Directo, Telefónica, Facebook, El Corte Inglés, and HP. May 12, 2016. The CNIO hosted an exhibition enti- tled ‘ Marie Sklodowska-Curie. A Pole in Paris ’, which revisited the personal and professional life of this key wom- an of the 20th century. This initiative of the CNIO Women in Science Office ( WISE ) stands testament to CNIO’s commitment to promoting and up- holding the work of women scientists. November 2016. CNIO’s Director, Maria A. Blasco ( left ), and the President of the CLH Group, José Luis López de Silanes ( right ), signed an agreement to foster collab- oration between both entities. Through this collaboration, the company joined ‘ CNIO Friends ’ in order to support research and training of research per- sonnel, with the goal of advancing the development of innovative and spe- cific therapies against cancer. March 18, 2016. ANNUAL REPORT 2016 186 International Affairs SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 187 ANNUAL REPORT 2016 188 INTERNATIONAL AFFAIRS INTERNATIONAL AFFAIRS CAROLINA POLA Director of International Affairs SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 189 The year 2016 marks the creation of the Department of International Affairs ( IAs ) at the CNIO ; its setup underscores the Centre’s commitment with increasing its international impact to continue its growth in scientific and professional excellence. The overarching goal of the Department is to provide a strategic global vision for the CNIO to facilitate the coordination of efforts from the different departments with the aim of substantially strengthening our international reach with the research community, policy makers and society. The lines of work initiated this year will enable us to reinforce four of CNIO’s fundamental concepts : knowledge sharing, value creation, research and innovation advancement, and professional and academic development. As an integral part of the CNIO, we strive to consolidate our Centre’s institutional leadership and reputation abroad, as well as to establish new robust partnerships and participate in European and International projects. This vision is now framed in the concept of ‘ Responsible Research & Innovation ( RRI )’ that will help us to further increase our impact on society by contributing to scientific advances and by endorsing innovative science policies. The department focuses on the continuation of existing CNIO initiatives and on propelling novel projects abroad. One of our key activities for the upcoming year is the co-organisation of an Innovative Medicine Initiative ( IMI ) Oncology Workshop that will be hosted at the CNIO and spearheaded by the CDTI ; its mission is to coordinate national efforts in upcoming cancer calls of this European public-private initiative. In this same vein, we have established a strategic workflow to coordinate CNIO’s efforts, keep investigators informed and facilitate their participation and leadership in H2020 projects—an effort to help CNIO researchers to reinforce their scientific potential and strengths in order to become even more competitive in the international scene. We have already sown the seeds for exciting alliances with international research centres of excellence that will pan out throughout 2017, not only in Europe but also in other countries where we can complement and synergise in regards to science, innovation and training. The CNIO will be formulating new ways of collaboration to continue attracting international talent and to provide our investigators with the most exciting, diverse and global setting for their professional and scientific growth. “ Broadening our reach and scientific impact through international collaborations is crucial for the CNIO investigators. We will continue consolidating our presence abroad with new initiatives.” ANNUAL REPORT 2016 190 CNIO Offices SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 191 Dean’s Office 192 CNIO Women In Science Office 194 ANNUAL REPORT 2016 192 CNIO OFFICES DEAN’S OFFICE MARÍA S. SOENGAS Dean for Academic Affairs Participants Mónica Álvarez, Ana F. Batalha, Hugo Bernard, Jasminka Boskovic, Bruna Calsina, Daniela Cerezo, Almudena Chaves, Guillermo de Cárcer, Lucía T. Díez, Silvia Janeiro, Eleonora Lapi, Ana Losada, Jorge Martínez, Raúl Martínez, David Olmeda, Laura Remacha, Maria Rigau, Federica Schiavioni The CNIO is recognised for the relevance and international projection of our Scientific Programmes. Key to this success is a solid core of undergraduate students, predoctoral and postdoctoral fellows, medical residents and a broad spectrum of visiting scientists. In fact, personnel in training constitute over 60% of the workforce in our institute. As such, the CNIO dedicates particular emphasis to career development, supported in part by highly competitive PhD and Postdoctoral Programmes. Agreements are also in place with multiple universities and medical centres, to ultimately bridge the gap between academic and clinical environments. Also very successful are our undergraduate summer internships, as well as diverse exchange and visitor programmes. Ultimately, our mission is to nurture and foster the development of our scientists-in-training, in order to maximise their chances of success. The CNIO Student Association ( CNIOSA ) and Postdoc Association ( CNIOPDA ) are the driving forces behind an inspiring series of seminars and workshops that we hold throughout the year. In this context, scientific reasoning, grant writing, manuscript organisation and CV preparation are just some of the many topics covered in our curriculum. Likewise, we acknowledge that career options extend beyond the bench and we therefore pay special attention to the areas of public communication, management of intellectual property, and the creation of start-ups or spin offs. These activities are performed in concert with CNIO’s Training Programmes and the Innovation and Communication Offices that are both deeply committed to providing the best environment for our personnel. We are most grateful to the Fundación Jesús Serra, for their continuous support to strengthen career development programmes at the CNIO. We believe that an informed society is better prepared to understand ( and if needed, face ) the diseases that constitute human cancer. Therefore, we are actively involved in SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 193 knowledge dissemination. Members of CNIOSA and CNIOPDA have participated in various school visits and Open Doors activities such as the Semana de la Ciencia or the European Researchers’Night ; all these events were highly attended, attracting over 250 participants of all ages. A particularly inspirational event this year was our Annual CNIO Lab Day. We were fortunate to host Simon Gifford, co-founder of Mashauri Limited, Director of Genesis Management Consulting and Professor at the prestigious Instituto de Empresa. Gifford gave an inspiring talk on how MBAs and entrepreneurs think and behave differently. He also spoke about his personal experience in setting up various consulting companies, emphasising the value of risk-taking and independent thinking. We also had seven outstanding talks given by CNIO trainees that covered exciting discoveries in the fields of epidemiology, epigenetics, proteomics, metastasis and drug development. Progress made in other basic and translational aspects of cancer were discussed in over sixty posters, which together emphasised the breadth of research covered by our different Scientific Programmes. Another main highlight of the Lab Day was the announcement of the recipients of our ‘ Director’s List Awards ’. These are recognitions of outstanding contributions made by our personnel in 3 categories : ( 1 ) predoctoral fellows with publications of the highest scientific impact ; ( 2 ) excellence in research by postdoctoral and staff investigators ; and ( 3 ) altruistic volunteering to further the mission of the Centre related to training, scientific divulgation and outreach. 1. Awards for Excellence in Research by Predoctoral Fellows Early in the year, the family Agüera-Nieto contacted the CNIO about giving a donation to support research conducted by young scientists. This led to the inauguration of the ‘ Antonia Nieto Award ’, in honour of their mother, to recognise the PhD student authoring the article with the highest impact. In 2016, the award went to Lluc Mosteiro for her impressive work published in Science on the mechanisms of cellular reprogramming in vivo. We are still moved by the kindness and commitment of the Agüera- Nieto family, who came to the CNIO to hand over this award and to share their multiple ideas and suggestions for the Amigos del CNIO initiative. Their energy is yet an additional inspiration for us to further strive to make significant contributions towards the advancement of cancer research. Additional awards in the PhD category were for Ana Teijeiro ( Cancer Cell ), Cristina Mayor-Ruiz ( Mol Cell ), Juan J. Montero ( Nat Commun ) and Eva Pérez-Guijarro ( Nat Commun ). 2. Award for Excellence in Research by Postdoctoral/ Staff Investigators The awardee was Matilde Murga, for her outstanding contributions to the fields of DNA replication and genomic instability ( Mol Cell and additional coauthored papers in Nat Struct and Mol Biol, Oncotarget and Sci Signaling ). 3. Outstanding Contribution to Outreach and Awareness This year’s recipient was Guillermo de Cárcer, for his tireless efforts in the organisation of the European Researcher’s Night. This is an event for which he coordinated over 50 volunteers, resulting in a flawless and exciting open-doors activity. The award was presented by Marcos Argumosa, who is himself an utterly impressive example of altruism, having run 10 consecutive marathons in 2015 to support the Amigos del CNIO initiative. In summary, we are as proud as ever of the achievements accomplished by our vibrant community of young investigators at the CNIO. We thank all those public and private contributors that have helped to support and fuel their efforts, and will make sure that the next years will be even more successful in moving the cancer field forward in a meaningful manner for the patients. “ At the CNIO we aim high : to carry out the most innovative and competitive basic and translational research, and to best prepare our trainees for the future, so that they can fulfil their potential as influential leaders.” DEAN’S OFFICE ANNUAL REPORT 2016 194 CNIO OFFICES CNIO WOMEN IN SCIENCE OFFICE Lola Martínez Coordinator Members Marinela Méndez : Work-Life Balance Coordinator, Francisca Mulero : Seminars and Events Coordinator, The CNIO Women in Science Office ( WISE ) was created at the end of 2012 with the aim to give visibility to women, to promote institutional awareness on gender equality, and to try to promote and support women in their professional careers. The WISE Office is composed of CNIO volunteers from different areas of the Centre, people who believe there is still a real need for action to be undertaken to ensure gender equality in the research career. Recent studies from different organisations in Spain and the European Union still display the typical ‘ scissors ’ graphic regarding the distribution of gender along the career ladder, together with an approximate 20% salary gap between men and women. Furthermore, women are still underrepresented as recipients of prestigious scientific awards ; as an example, another year has gone by without a single Nobel Prize, in any scientific discipline, being awarded to a woman. The issue does not seem to be that women are not present in academia ; the latest data show that 55% of women pursue university studies. Although it is worrying to see the lack of female students in the so-called ‘ STEM ’ careers, they are still well represented at the pre- and post-doctoral stages. However, this representation drops to a meagre 25% of women at the Principal Investigator level, and it is even lower at the levels of Department Directors and beyond. When experts analyse the data and try to identify the causes, one factor that keeps coming up repeatedly is the existence of cultural stereotypes between men and women. Since those stereotypes need to be challenged at an early age, the Office has decided to, from 2016 onwards, open up its seminars to schools and high schools with 2 main purposes : to make teenagers aware of those gender issues, and to promote equality and hopefully vocations among young girls to enter the scientific career. We are certain that within the CNIO community, we all need to continue working together, with the other CNIO administrative offices as well as the Works Council, in order to reach our common SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 195 CNIO WOMEN IN SCIENCE OFFICE goal, namely, to maintain the CNIO’s high level of scientific productivity and to ultimately make it an outstanding centre to work where gender barriers are completely eliminated and there is a sensible balance between work and life. The Office consists of two working groups : ɗ Work/Life Balance – aimed to promote and support initiatives to help improve the delicate balance between professional and personal life at the CNIO. ɗ Seminars and Events – aimed to raise awareness of gender issues, and provide networking opportunities to all CNIO researchers. In 2016, the WISE Office was again able to invite and welcome several top female leaders from different areas to tell us about their experience with gender issues, giving our young scientists ideas and advice on how to best overcome some of the hurdles that they may face during their careers, while also giving CNIO researchers the opportunity to expand their networks. Some of the seminars organised by the WISE Office during 2016 are : ɗ EXPEDITION TO SUCCESS : Achieving goals and overcoming difficulties. Edurne Pasaban. Mountaineer, Tolosa, Spain. March 8th. ɗ Recuerdos y Olvidos Feministas. María Teresa Fernández de la Vega. President of the Women for Africa Foundation, former Vice President of Spain. May 10th. ɗ Las Sinsombrero, sin ellas la historia no está completa. Tánia Balló. Documentalist. Barcelona, Spain. June 28th. ɗ Marie Skłodowska-Curie : Medical Physics pioneer and inspiration to female scientists. Dr Guadalupe Martín Martín. Medical Physicist. Fuenlabrada University Hospital. Madrid, Spain. November 7th. Also, in November, we hosted the itinerant exhibition on the life and work of two-time Nobel Laureate, Marie Sklodowska-Curie, who is still an inspiration to women scientists. We put together six organised visits to the exhibition combined with a tour to different CNIO laboratories for students and the general public ; these were a great success. Since 1901, the year when Alfred Nobel established the awards bearing his name, only 17 women have been honoured with the recognition in the scientific field and unfortunately, many were left out despite their now widely recognised merits. We wish to contribute towards changing those numbers, and so we are working on developing different mentorship and leadership programmes for CNIO researchers. “ Within the CNIO community, we continue to work together towards the elimination of gender barriers, the empowerment of women in Science and society and the promotion of scientific vocations among young girls. Scientific Excellence can only be fully achieved through gender equality.” Albanderi Alfraidi, Maria A. Blasco, Nicole Dölker, Raquel García-Medina, Francesc Madriles, Alba de Martino, Cristina de Martos (since November), Diego Megías, Fernando Peláez, Carolina Pola (since November), Alejandra Tavera ANNUAL REPORT 2016 196 Facts & Figures SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 197 Scientific Management 198 Competitive Funding 200 Education and Training Programmes 214 Scientific Events 220 Administration 234 Board of Trustees 234 Scientific Advisory Board 236 Management 238 CNIO Personnel 2016 240 ANNUAL REPORT 2016 198 FACTS & FIGURES SCIENTIFIC MANAGEMENT ISABEL BARTHELEMY Scientific Management Director Raquel Ares, Sonia Cerdá, Almudena del Codo ( since March ), M. Dolores Liébanes, Victoria López, Ana Merino, Juan Ramón Molina, Mercedes Moro, Leyre Vergés ( since December ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 199 SCIENTIFIC MANAGEMENT | The Scientific Management Department at the CNIO is committed to assisting with the facilitation of all those key areas that help our scientists to better focus their efforts on their research. The Department encompasses various Offices : Projects and Consortia, Education and Training Programmes, Scientific Events, Scientific Publishing, and Library & Archives. The mission of the Projects’ Office is to guide the CNIO scientists through all stages related to the application and management processes of externally-funded projects, whether they be financed through either public and/or private institutions, or stem from either national or international funding bodies. The Office coordinates the internal call alerts, manages the ethical certification for projects involving animal experimentation or human samples, supports scientists with the preparation of the project proposals, manages the ongoing projects, and contacts the funding agencies to resolve any issues or deal with questions. The Training Office is the central point for training at the CNIO ; it aids the recruitment process, serves as an advocate for all fellows, provides administrative support, and creates educational and learning opportunities. It is responsible for helping PhD students, postdoctoral scientists and post-resident MDs by announcing call alerts and providing the relevant key information ; helping foreign students with their paperwork at the foreign office ; organising the summer training call ; and, in general, in collaboration with the Personnel Department, managing student’s grants. The Events Office organises CNIO meetings, such as the CNIO Frontiers Meetings, the Distinguished Seminars series, the external Scientific Advisory Board ( SAB ) meeting, CNIO Progress Reports, as well as Faculty retreats, among others. The Office also helps scientists by providing advice for the organisation of specific events, including scientific divulgation events, and arranges the CNIO guided visits. The Publications Office is responsible for the preparation of institutional scientific publications, including the CNIO Annual Report, booklets of the Scientific Advisory Board meeting and those of other symposia, as well as scientific dissemination leaflets. The Office also provides support for the scientific editing of press notes and other publications of scientific divulgation to a non-specialised audience. The Library administers the electronic subscriptions of over 300 scientific journals at the CNIO and manages journal article requests for journals that the CNIO is not subscribed to. The Library also provides information regarding reference management software. “ Every day, we work towards building a strong and flexible framework to support our scientists and to help them achieve excellence.” ANNUAL REPORT 2016 200 FACTS & FIGURES COMPETITIVE FUNDING The CNIO attracts a substantial proportion of its funding from external sources. Most of this funding comes from national and international funding bodies. In 2016, researchers at the CNIO were involved in 148 projects that received extramural funding. CNIO actively participates 64 collaborative projects in total : 30 were international collaborative projects ( 4 of which are coordinated by the CNIO ) and 34 collaborative projects at the national level ( 12 of them coordinated by the CNIO ). The international collaborative projects were funded by institutions such as the European Commission through the 7 Framework Programme and Horizon 2020, the Interreg SUDOE Programme, the US National Institutes of Health ( NIH ), the US Department of Defense ( DoD ), the Melanoma Research Alliance, the Paradifference Foundation, the Worldwide Cancer Research and the Volkswagen foundation In addition to these collaborative projects, researchers at the CNIO attracted funding for projects carried out by individual groups. In 2016, 21 of these projects received international funds while 63 of them received national funding ( mainly the Spanish Ministry of Economy, Industry and Competitiveness and the Institute of Health Carlos III ). The international individual projects are funded by the European Commission ( European Research Council ( ERC ) grants and the Marie Curie Actions ), the Worldwide Cancer Research ( WCR ), the Howard Hughes Medical Institute ( HHMI ) and the European Foundation for the Study of Diabetes ( EFSD ). SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 201 SCIENTIFIC MANAGEMENT | COMPETITIVE FUNDING INTERNATIONAL GRANTS COLLABORATIVE PROJECTS EUROPEAN COMMISSION 7TH FRAMEWORK PROGRAMME ( 2007-2013 ) COST ACTION PRINCIPAL INVESTIGATOR PROJECT TITLE Malats, Núria ( coordinator ) COST Action EU Pancreas : An integrated European platform for pancreas cancer research : from basic science to clinical and public health interventions for a rare disease ( Ref.: COST BM1204 ) EURATOM PRINCIPAL INVESTIGATOR PROJECT TITLE Serrano, Manuel RISK-IR : Risk, Stem Cells and Tissue Kinetics-Ionising Radiation ( Ref.: 323267 ) INNOVATIVE MEDICINES INITIATIVE JOINT UNDERTAKING ( IMI JU ) PRINCIPAL INVESTIGATOR PROJECT TITLE Valencia, Alfonso e-TOX : Integrating bioinformatics and chemoinformatics approaches for the development of expert systems allowing the in silico prediction of toxicities ( Ref.: 115002 ) Valencia, Alfonso Open PHACTS : An open, integrated and sustainable chemistry, biology and pharmacology knowledge resource for drug discovery ( Ref.: 115191-2 ) INTEGRATED PROJECT ( IP ) PRINCIPAL INVESTIGATOR PROJECT TITLE Valencia, Alfonso BLUEPRINT : A BLUEPRINT of haematopoietic epigenomes ( Ref.: 282510 ) Valencia, Alfonso ASSET : Analysing and striking the sensitivities of embryonal tumours ( Ref.: 259348 ) Valencia, Alfonso RD-CONNECT : An integrated platform connecting registries, biobanks and clinical bioinformatics for rare disease research ( Ref.: 305444 ) MARIE CURIE ACTIONS ( MCA ) PRINCIPAL INVESTIGATOR PROJECT TITLE Fernández-Capetillo, Óscar ITN aDDRess : Joint training and research network on chromatin dynamics and the DNA damage response ( Ref.: 316390 ) NETWORKS OF EXCELLENCE ( NOE ) PRINCIPAL INVESTIGATOR PROJECT TITLE Barbacid, Mariano EUROCANPLATFORM : A European platform for translational cancer research ( Ref.: 260791 ) ANNUAL REPORT 2016 202 FACTS & FIGURES SMALL OR MEDIUM-SCALE FOCUSED RESEARCH PROJECTS PRINCIPAL INVESTIGATOR PROJECT TITLE Malats, Núria TransBioBC : Translation of novel Biomarkers for Bladder Cancer for clinical outcome prediction ( Ref.: 601933 ) Robledo, Mercedes ENS@T- CANCER : European network for the study of adrenal tumours- structuring clinical research on adrenal cancers in adults. ( Ref.: 259735 ) ERA-NET ON TRANSLATIONAL CANCER RESEARCH ( TRANSCAN ) PRINCIPAL INVESTIGATOR PROJECT TITLE Malats, Núria Bio-PaC : Biomarkers of tumor recurrence in pancreatic cancer ( financed by ISCIII, Ref.: AC14/00025 ) ERA NET NEURON II : NETWORK OF EUROPEAN FUNDING FOR NEUROSCIENCE RESEARCH PRINCIPAL INVESTIGATOR PROJECT TITLE Malumbres, Marcos MicroKin : Deciphering the multifaceted pathways underlying MCPH pathogenesis in the mouse and human ( Financed by MEIC Ref.: PCIN-2015-007 ) HORIZON 2020 ( 2014-2020 ) RESEARCH INFRASTRUCTURES, INCLUDING E-INFRASTRUCTURES PRINCIPAL INVESTIGATOR PROJECT TITLE Valencia, Alfonso ELIXIR-EXCELERATE : Fast-track ELIXIR implementation and drive early user exploitation across the life-sciences ( Ref.: 676559 ) Valencia, Alfonso OpenMinTeD : Mining INfrastructure for TExt and Data ( Ref.: 654021 ) MARIE SKŁODOWSKA-CURIE ACTIONS ( MSCA ) PRINCIPAL INVESTIGATOR PROJECT TITLE Soengas, María S. ITN IMMUTRAIN : Training network for the immunotherapy of cancer ( Ref.: 641549 ) SOCIETAL CHALLENGE 1 : HEALTH, DEMOGRAPHIC CHANGE AND WELLBEING PRINCIPAL INVESTIGATOR PROJECT TITLE Benítez, Javier BRIDGES : Breast cancer risk after diagnostic gene sequencing ( Ref.: 634935 ) INDUSTRIAL TECHNOLOGIES : ADVANCED MATERIALS AND NANOTECHNOLOGIES PRINCIPAL INVESTIGATOR PROJECT TITLE Hidalgo, Manuel NoCanTher : Nanomedicine upscaling for early clinical phases of multimodal cancer therapy ( Ref. :685795 ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 203 SCIENTIFIC MANAGEMENT | COMPETITIVE FUNDING INTERREG SUDOE PROGRAMME 1 PRINCIPAL INVESTIGATOR PROJECT TITLE Valencia, Alfonso European Network for Translational Research and Innovation in Oncology /Réseau Européen de Recherche translationnelle et d’Innovation en oncologie ( Ref.: SOE1/P1/F0082 ) MELANOMA RESEARCH ALLIANCE ( MRA ) PRINCIPAL INVESTIGATOR PROJECT TITLE Peinado, Héctor ; Soengas, María S. ( coordinator ) Imaging and therapeutic targeting of lymphangiogenesis in melanoma ( Ref.: 269626 ) Soengas, María S. ( coordinator ) Imaging and targeting dormant and pro-metastatic melanoma lesions in vivo ( Ref.: 401181 ) THE PARADIFFERENCE FOUNDATION PRINCIPAL INVESTIGATOR PROJECT TITLE Robledo, Mercedes SDHB-related metastatic paraganglioma : search for the cure US CONGRESSIONALLY DIRECTED MEDICAL RESEARCH PROGRAMS ( CDMRP )/US DEPARTMENT OF DEFENSE PRINCIPAL INVESTIGATOR PROJECT TITLE Peinado, Héctor Radiolabeled exosomes for the early detection of metastases and to predict breast cancer premetastatic niche ( Ref.: W81XWH-13-1-0249 ) Peinado, Héctor Organ-tropic metastatic secretomes and exosomes in breast cancer ( Ref.: W81XWH-13-1-0427 ) Peinado, Héctor Exosomes in Development and Therapy of Malignant Mesothelioma ( Ref.: W81XWH-14-1-0199 ) US NATIONAL INSTITUTES OF HEALTH ( NIH ) PRINCIPAL INVESTIGATOR PROJECT TITLE Peinado, Héctor Characterization and functional analysis of breast cancer secreted exosomes in malignant progression ( Ref.: U01CA169538 ) Peinado, Héctor Exosome-mediated transfer of c-MET to bone marrow progenitors promotes metastasis ( Ref.: R01CA169416 ) Valencia, Alfonso GENCODE 2 : Integrated human genome annotation : generation of a reference gene set ( Ref.: HG007234-01 ) VOLKSWAGEN FOUNDATION PRINCIPAL INVESTIGATOR PROJECT TITLE Lietha, Daniel Nanoapertures loaded with individual molecules ( Ref.: 86416-1 ) WORLDWIDE CANCER RESEARCH ( WCR, FORMERLY AICR ) PRINCIPAL INVESTIGATOR PROJECT TITLE Malats, Núria ( coordinator ) Oral microbiotic profiles and its association with risk of pancreatic ductal adenocarcinoma ( Ref.: 15-0391 ) 1. This Programme is cofunded by the European Regional Development Fund ( ERDF )  ANNUAL REPORT 2016 204 FACTS & FIGURES INTERNATIONAL GRANTS INDIVIDUAL PROJECTS EUROPEAN COMMISSION 7TH FRAMEWORK PROGRAMME ( 2007-2013 ) EUROPEAN RESEARCH COUNCIL ( ERC ) PRINCIPAL INVESTIGATOR PROJECT TITLE Fernández-Capetillo, Óscar ERC Consolidator Grant RSHEALTH : Investigating the causes and consequences of replication stress in mammalian health ( Ref.: 617840 ) MARIE CURIE ACTIONS ( MCA ) PRINCIPAL INVESTIGATOR PROJECT TITLE Al-Shahrour, Fátima PERSMEDOMICS : Bioinformatics and integrative genomics for a novel personalized cancer therapy ( Ref.: 334361 ) Peinado, Héctor WHRI COFUND ADIPOMET : Analyzing the crosstalk of tumor and adipose tissue during metastasis ( Ref.: 608765 ) Ramón, Santiago; Moreno, María WHRI COFUND CAD_FL : Revealing the functional mechanism of CAD and its potential as a therapeutic target ( Ref.: 608765 ) Squatrito, Massimo GLIDD : DNA Damage Response ( DDR ) signaling in tumor formation and therapeutic resistance of gliomas ( Ref.: 618751 ) Wagner, Erwin F.; Gago, Nuria WHRI COFUND STEM-PSO : Unraveling the contribution of Epidermal and Non-Epidermal Progenitor ( Ref.: 608765 ) HORIZON 2020 ( 2014-2020 ) EUROPEAN RESEARCH COUNCIL ( ERC ) PRINCIPAL INVESTIGATOR PROJECT TITLE Efeyan, Alejo ERC Starting Grant NutrientSensingVivo : The Physiology of Nutrient Sensing by mTOR ( Ref.: 638891 ) Hidalgo, Manuel ERC Advanced Grant AVATAR : Integrating Genomics and Avatar Mouse Models to Personalize Pancreatic Cancer Treatment ( Ref.: 670582 ) Serrano, Manuel ERC Advanced Grant CELLPASTICITY : New Frontiers in Cellular Reprogramming : Exploiting Cellular Plasticity ( Ref.: 669622 ) EUROPEAN FOUNDATION FOR THE STUDY OF DIABETES ( EFSD ) PRINCIPAL INVESTIGATOR PROJECT TITLE Djouder, Nabil Growth factors and nutrients in type 2 diabetes : role of URI in β cell plasticity and glucose homeostasis HOWARD HUGHES MEDICAL INSTITUTE ( HHMI ) PRINCIPAL INVESTIGATOR PROJECT TITLE Fernández-Capetillo, Óscar Exploring the role of replicative stress in cancer and ageing ( Ref.: 55007417 ) MELANOMA RESEARCH ALLIANCE ( MRA ) PRINCIPAL INVESTIGATOR PROJECT TITLE Soengas, María S. Prognostic and therapeutic impact of lymphovascular niches in melanoma ( Ref.: 348673 ) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 205 SCIENTIFIC MANAGEMENT | COMPETITIVE FUNDING PROSTATE CANCER FOUNDATION PRINCIPAL INVESTIGATOR PROJECT TITLE Olmos, David Integration of clinical, molecular and biological characteristics to define an aggressive subtype of prostate cancer based on deficient homologous recombination WORLDWIDE CANCER RESEARCH ( WCR, FORMERLY AICR ) PRINCIPAL INVESTIGATOR PROJECT TITLE Blasco, Maria Targeting telomeres in cancer ( Ref.: 16-1177 ) Lietha, Daniel Targeting regulatory mechanisms for allosteric cancer drug discovery ( Ref.: 15-1177 ) Malumbres, Marcos New therapeutic strategies by inhibiting Mastl in breast tumors ( Ref.: 15-0278 ) Peinado, Héctor Evaluation of obesity as a novel risk factor in metastasis ( Ref.: 16-1244 ) Pérez Moreno, Mirna A. Defining the role of macrophage-derived Wnts in squamous cell carcinoma ( Ref.: 15-1219 ) Soengas, María S. Harnessing endo/exocytosis for a coordinated targeting of melanoma cells, their vasculature and the immune system ( Ref.: 15-1374 Wagner, Erwin F. Dissecting the roles of Fra proteins in lung adenocarcinoma progression and metastasis ( Ref.: 13-0216 ) US CONGRESSIONALLY DIRECTED MEDICAL RESEARCH PROGRAMS ( CDMRP )/US DEPARTMENT OF DEFENSE PRINCIPAL INVESTIGATOR PROJECT TITLE Peinado, Héctor Role of exosomes and Endoglin in Neurofibromatosis Progression ( Ref.: W81XWH-16-1-0131 ) ANNUAL REPORT 2016 206 FACTS & FIGURES NATIONAL GRANTS COLLABORATIVE PROJECTS COMMUNITY OF MADRID / COMUNIDAD AUTÓNOMA DE MADRID 2 PRINCIPAL INVESTIGATOR PROJECT TITLE Barbacid, Mariano ; Malumbres, Marcos ( coordinator ) Programa ONCOCYCLE : El ciclo celular y los microRNAs en la autorenovación y diferenciación de células progenitoras ( Ref.: S2011/BMD-2470 ) Blasco, Maria A. ; Serrano, Manuel ( coordinator ) Programa ReCaRe : Reprogramación en cáncer y regeneración ( Ref.: S2011/BMD-2303 ) Campos-Olivas, Ramón; Lietha, Daniel Programa BIPEDD 2 : Plataforma integrada de bioinformática para el descubrimiento de nuevos fármacos basado en la estructura del receptor ( Ref.: S2011/BMD-2457 ) González-Neira, Anna Programa VISIONANIMAL : Modelos animales para el estudio de enfermedades de la visión ( Ref.: S2011/BMD-2439 ) Martínez, Jorge L. Programa ANGIOBODIES 2 : Desarrollo de anticuerpos recombinantes para uso terapéutico y diagnostico en angiogénesis patológica y para la identificación de nuevos marcadores angiogénicos ( Ref.: S2011/BMD-2312 ) Montoya, Guillermo Programa INTERACTOMICS : Interactomics del centrosoma ( Ref.: S2011/BMD-2305 ) Robledo, Mercedes Programa TIRONET : Fisiopatología tiroidea : Mecanismos implicados en cáncer, autoinmunidad y mecanismo de acción de hormonas tiroideas ( Ref.: S2011/BMO-2328 ) Soengas, María S. Programa NANODENMED : Nanosistemas dendríticos como agentes y vectores terapéuticos en distintas aplicaciones biomédicas ( Ref.: S2011/BMD-2351 ) Real, Francisco X. Programa CEL-DD : Linajes y competición celular en el desarrollo y la enfermedad ( Ref.: P2010/BMD-2315 ) INSTITUTE OF HEALTH CARLOS III / INSTITUTO DE SALUD CARLOS III ( ISCIII ) SUB-PROGRAMME OF COOPERATIVE HEALTH RESEARCH THEMATIC NETWORKS/SUBPROGRAMA DE REDES TEMÁTICAS DE INVESTIGACIÓN COOPERATIVA ( RETICS ) 3 PRINCIPAL INVESTIGATOR PROJECT TITLE Cigudosa, Juan C. Red Temática de Investigación Cooperativa en Cáncer ( RTICC ) ( Group RD12/0036/0037 ) Malats, Núria Red Temática de Investigación Cooperativa en Cáncer ( RTICC ) ( Group RD12/0036/0050 ) Real, Francisco X. Red Temática de Investigación Cooperativa en Cáncer ( RTICC ) ( Group RD12/0036/0034 ) SUB-PROGRAMME OF GRANTS FOR RESEARCH SUPPORT PLATFORMS IN HEALTH SCIENCES AND TECHNOLOGY/ SUBPROGRAMA DE AYUDAS PARA PLATAFORMAS DE APOYO A LA INVESTIGACIÓN EN CIENCIAS Y TECNOLOGÍAS DE LA SALUD 4 PRINCIPAL INVESTIGATOR PROJECT TITLE Benítez, Javier Plataforma de recursos biomoleculares y bioinformáticos, PRB 2 ( PT13/0001/0005 ) Morente, Manuel M. ( coordinator ) Plataforma de Biobancos ( Coordination node and group ( PT13/0010/0001 ) 2. This Programme is cofunded by European Structural and Development Funds ( ERDF and ESF )  3. This Programme is cofunded by the European Regional Development Fund ( ERDF )  SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 207 SCIENTIFIC MANAGEMENT | COMPETITIVE FUNDING Muñoz, Javier Plataforma de recursos biomoleculares y bioinformáticos, PRB 2 ( Group PT13/001/0010 ) Valencia, Alfonso Plataforma de recursos biomoleculares y bioinformáticos, PRB 2 ( Group PT13/0001/0030 ) RESEARCH PROJECTS IN HEALTH 5 PRINCIPAL INVESTIGATOR PROJECT TITLE Blasco, Maria A. Cellular aging in first episode early-onset psychosis. Collaboration with Gregorio Marañón Hospital ( Ref.: PI14/00397 ) Blasco, Maria A. Safety and efficacy of gene therapy with telomerase in acute myocardial infarction. Impact on ventricular remodeling in an experimental porcine model. Collaboration with Gregorio Marañón Hospital ( Ref. DTS15/00095 ) MINISTRY OF HEALTH, SOCIAL SERVICES AND EQUALITY / MINISTERIO DE SANIDAD, SERVICIOS SOCIALES E IGUALDAD ( MSSSI ) PRINCIPAL INVESTIGATOR PROJECT TITLE Hidalgo, Manuel ( coordinator ) Chemosensitivity profiles for the personalized therapy of advanced colorectal cancer ( Ref.: EC11-017 ) Hidalgo, Manuel ( coordinator ) Personalized treatment for pancreatic cancer patients ( Ref.: EC10-278 ) Hidalgo, Manuel ( coordinator ) Personalized treatment for pancreatic cancer patients II ( Ref.: EC11-005 ) MINISTRY OF ECONOMY, INDUSTRY AND COMPETITIVENESS / MINISTERIO DE ECONOMÍA, INDUSTRIA Y COMPETITIVIDAD ( MEIC ) NATIONAL PLAN FOR SCIENTIFIC AND TECHNICAL RESEARCH AND INNOVATION ( 2013-2016 ) EXCELLENCE NETWORKS / REDES DE EXCELENCIA PRINCIPAL INVESTIGATOR PROJECT TITLE Barbacid, Mariano ( coordinator ) ; Blasco, Maria A. ; Fernández-Capetillo, Óscar ; Malumbres, Marcos ; Real, Francisco X. ; Serrano, Manuel OncoBIO : Cancer biology ( Ref.: SAF2014-57791-REDC ) Malumbres, Marcos ( coordinator ) CellSYS : Functional and Systems Biology of Cell Proliferation ( Ref.: BFU2014-52125-REDT ) Serrano, Manuel ( coordinator ) SENESTHERAPY : Cell senescence in cancer therapy ( Ref.: SAF2014-56720-REDT ) CHALLENGES-COLLABORATION/RETOS-COLABORACIÓN 6 PRINCIPAL INVESTIGATOR PROJECT TITLE Djouder, Nabil NRCANCER- Desarrollo de nueva terapia antitumoral basada en nicotinamida-ribosido ( Ref.: RTC-2016-5431-1 ) Soengas, María S. Ensayo Clínico Fase I de BO-110 : un nuevo tratamiento para melanoma avanzado y otros tumores ( Ref.: RTC-2014-2442-1 ) 4. This Programme is cofunded by the European Regional Development Fund ( ERDF )  5. This Programme is cofunded by the European Regional Development Fund ( ERDF )  ANNUAL REPORT 2016 208 FACTS & FIGURES SPANISH ASSOCIATION AGAINST CANCER / ASOCIACIÓN ESPAÑOLA CONTRA EL CÁNCER ( AECC ) PRINCIPAL INVESTIGATOR PROJECT TITLE Barbacid, Mariano ( coordinator ) A multifaceted approach to target pancreatic cancer ( Ref.: GC16173694 ) Benítez, Javier Cancer and immunodeficiency in children ( Ref.: CEI14142070 ) Malats, Núria ; Real, Francisco X. ( coordinator ) Invasive bladder cancer : towards precision medicine ( Ref.: GCB14142293 ) González Pisano, David ; Peinado, Héctor ; Soengas, María S. ( coordinator ) Distinct routes of metastatic dissemination in different melanoma subtypes. Implications in the validation of new tumor biomarkers and therapeutic targets ( Ref.: GCB15152978 ) LA MARATÓ TV3 FOUNDATION / FUNDACIÓN LA MARATÓ TV3 PRINCIPAL INVESTIGATOR PROJECT TITLE Barbacid, Mariano Molecular analysis of Capicua, a novel tumor suppressor involved in RTK signaling and transcriptional repression ( Ref.: 20131730/31 ) Fernández-Capetillo, Óscar Exploring synthetic lethal interactions between PARP and the DNA damage response in cancer treatment ( Ref.: 20134130/31 ) Soengas, María S. Role of RNA binding proteins in melanoma progression : searching for new diagnostic markers and therapeutic targets ( Ref.: 20131430/31 ) MADRI+D FOUNDATION / FUNDACIÓN PARA EL CONOCIMIENTO MADRI+D PRINCIPAL INVESTIGATOR PROJECT TITLE Dean’s Office for Academic Affairs ; Soengas, María S. European Researchers ’ Night 2014, organized by Madri+d Foundation and founded by European Comission on the framework of H2020 Programme 6. This Programme is cofunded by the European Regional Development Fund ( ERDF )  SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 209 SCIENTIFIC MANAGEMENT | COMPETITIVE FUNDING NATIONAL GRANTS INDIVIDUAL PROJECTS INSTITUTE OF HEALTH CARLOS III / INSTITUTO DE SALUD CARLOS III ( ISCIII ) RESEARCH PROJECTS IN HEALTH 7 PRINCIPAL INVESTIGATOR PROJECT TITLE Benítez, Javier Biologic and genetic bases of telomere shortening in hereditary breast cancer. Searching for new high susceptibility genes in BRCAX families with short telomeres ( Ref.: PI12/00070 ) Cascón, Alberto Exome sequencing of trios, mother-father-proband, in pediatric patients with multiple pheochromocytomas/paragangliomas ( Ref.: PI12/00236 ) Cascón, Alberto Next generaton sequencing of genes directly and indirectly involved in the Krebs cycle, applied to pheochromocytomas/paragangliomas with hypermethylated phenotype ( Ref.: PI15/00783 ) Cigudosa, Juan C. Genetic diagnostics by next-generation-sequencing in myeloid neoplasias : step towards its clinical use and characterization studies on the mutation genomic and functional pathological effects ( Ref.: PI12/00425 ) García, María José Definition of novel ovarian cancer susceptibility genes using next- generation sequencing technology and a LOH-candidate region approach in high-risk non-BRCA1/BRCA2 patients ( Ref.: PI12/01319 ) González-Neira, Anna Personalizing breast cancer treatment : prediction model construction for taxanes and anthracyclines efficacy thought the integration of different genomic approaches ( Ref.: PI12/00226 ) Hidalgo, Manuel Targeting Pancreatic Cancer Stroma ( Ref.: PI13/00230 ) Malats, Núria Aetiology of pancreas cancer : Application of “ omics ” technologies in the assessment of risk factors ( Ref.: PI12/00815 ) Malats, Núria Building and validation of risk prediction models for pancreas cancer. The application of a multi-omics approach ( Ref.: PI15/01573 ) Molina, María Esther Dietary patterns, antioxidants and biomarkers of oxidant-antioxidant status in the EPIC-Granada and EPIC-Gipuzkoa ( European Prospective Investigation into Cancer and Nutrition ) cohort ( Ref.: PI12/00002 ) Pérez de Castro, Ignacio An integrative Study of Chromosomal Instability and Cancer : looking for prognostic markers and therapeutic opportunities ( Ref.: PI14/00227 ) Olmos, David Homologous recombination DNA repair deficiency related chromosomal instability in aggressive prostate cancer ( Ref.: PI13/01287 ) Quintela, Miguel Ángel From systems biology to clinical trials : high-throughput studies and definition of predictive factors and resistance mechanisms against breast cancer drugs ( Ref.: PI13/00430 ) Robledo, Mercedes Prognostic profiles in endocrine tumours identified by next generation sequencing, and definition of markers with clinical utility ( Ref.: PI14/00240 ) Rodríguez, Sandra Ewing Sarcoma Model : induction of the t( 11 ;22 ) translocation in human mesenchymal stem and iPS cells by the CRISPR-Cas9 system and study of the cellular context and other secondary events role ( Ref.: PI14/01884 ) Squatrito, Massimo Investigating the role of Fra1 and Fra2 in glioma tumor formation and treatment response ( Ref.: PI13/01028 ) Urioste, Miguel PTEN-hamartoma tumour syndrome research : Phenotypic spectrum, associated cancers, molecular basis and search of new gene ( Ref.: PI14/00459 ) 7. This Programme is cofunded by the European Regional Development Fund ( ERDF )  ANNUAL REPORT 2016 210 FACTS & FIGURES MINISTRY OF ECONOMY, INDUSTRY AND COMPETITIVENESS / MINISTERIO DE ECONOMÍA, INDUSTRIA Y COMPETITIVIDAD ( MEIC ) NATIONAL R&D&I PLAN 2008-2011 SUB-PROGRAMME FOR NON-TARGETED FUNDAMENTAL RESEARCH PROJECTS / SUBPROGRAMA DE PROYECTOS DE INVESTIGACIÓN FUNDAMENTAL NO ORIENTADA PRINCIPAL INVESTIGATOR PROJECT TITLE UluÇkan, Özge PsorTACEmiR21 : Investigating the role of microRNA21/ TIMP-3/TACE in psoriasis - evaluating the potential therapeutic implications ( Ref.: SAF2012-39670 ) Valencia, Alfonso Development of biocomputing systems and subjacent computational methods for the analysis of oncologic personalised therapies ( Ref.: BIO2012-40205 ) Wagner, Erwin F. HepAP-1 : From liver physiology to hepatitis and hepatocellular carcinoma ( HCC ): role of AP-1 ( Fos/Jun ) proteins ( Ref.: BFU2012-40230 ) NATIONAL PLAN FOR SCIENTIFIC AND TECHNICAL RESEARCH AND INNOVATION ( 2013-2016 ) CENTRES AND UNITS OF EXCELLENCE “ SEVERO OCHOA ” SUB-PROGRAMME / SUBPROGRAMA DE APOYO A CENTROS Y UNIDADES DE EXCELENCIA ‘ SEVERO OCHOA ’ PRINCIPAL INVESTIGATOR PROJECT TITLE Blasco, Maria A. Center of Excellence “ Severo Ochoa ” ( Ref.: SEV-2015-0510 ) R&D EXCELLENCE PROJECTS / PROYECTOS DE I+D EXCELENCIA 8 PRINCIPAL INVESTIGATOR PROJECT TITLE Méndez, Juan REPLICON : Molecular mechanisms that control eukaryotic DNA replication ( Ref.: BFU2013-49153-P ) Ramón, Santiago CADstructure : Structural determination of the architecture of CAD, an antitumoral target that controls the biosynthesis of pyrimidines ( Ref.: BFU2013-48365-P ) Ruiz, Sergio RSHIPS : Replicative stress during somatic cell reprogramming ( Ref.: SAF2013-49147-P ) CHALLENGES-RESEARCH /RETOS-INVESTIGACIÓN 9 PRINCIPAL INVESTIGATOR PROJECT TITLE Barbacid, Mariano PANTHER : A three prong strategy to fight pancreatic ductal adenocarcinoma ( Ref.: SAF2014-59864-R ) Blasco, Maria A. TeloHealth : Telomeres, telomerase and disease ( Ref.: SAF2013-45111-R ) Djouder, Nabil MILC : Metabolic inflammation in liver cancer ( Ref.: SAF2013-46089-R ) Efeyan, Alejo NUTRIENTOR : Physiology of nutrient sensing and signaling by the mTOR complex 1 ( Ref.: SAF2015-67538-R ) Fernández-Capetillo, Óscar BREAKINGRAD : Exploring the limits of radioresistance in mammals ( Ref.: SAF2014-59498-R ) Losada, Ana COHESIN : Cohesin function and regulation : a multidisciplinary approach ( Ref.: BFU2013-48481-R ) 8. This Programme is cofunded by the European Regional Development Fund ( ERDF )  9. This Programme is cofunded by the European Regional Development Fund ( ERDF )  SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 211 SCIENTIFIC MANAGEMENT | COMPETITIVE FUNDING Malumbres, Marcos Cyclexit : Physiological and therapeutic relevance of mitotic kinases and phosphatases ( Ref.: SAF2015-69920-R ) Muñoz, Daniel REMODEL : Cellular senescence as an active player in tissue remodeling ( Ref.: BFU2014-60020-R ) Muñoz, Javier steMS : Understanding ground state pluripotency of embryonic stem cells through mass spectrometry-based proteomics ( Ref.: SAF2013-45504-R ) Ortega, Sagrario HaploEScancer : Haploid ES cells for cancer research ( Ref.: SAF2013-44866-R ) Osorio, Ana IPAGEN : Exploring the mechanism of action of PARP inhibitors in breast and ovarian cancer patients. Identification of new genetic predictors of response ( Ref.: SAF2014-57680-R ) Pastor, Joaquín CDK8eDD : CDK8 a novel target in cancer therapy. Relevance of CDK8 kinase activity, discovery and optimization of selective orally bioavailable CDK8 inhibitor ( Ref.: SAF2013-44267-R ) Peinado, Héctor METASTAXOMEs : Role of tumor-secreted exosomes in lymph node microenvironment reprogramming during metastasis ( Ref.: SAF2014-54541-R ) Pérez Moreno, Mirna A. ESSENCE : Extrinsic control of the skin stem cell niche in homeostasis and cancer ( Ref.: BFU2015-71376-R ) Real, Francisco X. TRANS-PDAC : Transcriptional control of pancreatic cancer development ( Ref.: SAF2015-70553-R ) Rodríguez, Cristina PREDICT : Identification of genetic markers and physiopathologic factors predictive of the peripheral neuropathy of paclitaxel and of other oncologic drugs : massive sequencing of candidate genes ( Ref.: SAF2015-64850-R ) Serrano, Manuel CANCERAGE : Cancer and ageing-associated diseases : new frontiers and new strategies ( Ref.: SAF2013-48256-R ) Soengas, María S. MEL-STOP : Vesicular trafficking in melanoma progression and treatment response ( Ref.: SAF2014-56868-R ) Valiente, Manuel ReACTIVE BrainMET : Dissecting the role of reactive astrocytes in brain metastasis ( Ref.: SAF2014-57243-R ) Valencia, Alfonso EPIC : Expression Patterns of Inverse Comorbidity ( Ref.: BFU2015-71241-R ) Wagner, Erwin F. CANPSOR : Investigating Cancer Risk in Psoriasis ( Ref.: SAF2015-70857-R ) EXCELLENCE-EUROPE / EUROPA EXCELENCIA PRINCIPAL INVESTIGATOR PROJECT TITLE Rodríguez, Cristina ANGIOMARKER : Predicting antiangiogenic drug response in cancer : markers and mechanisms ( Ref.: SAF2015-70820-ERC ) Valiente, Manuel BrainMET : Deconstructing metastatic disease in the brain ( Ref.: SAF2015-62547-ERC ) RESEARCH-EUROPE / EUROPA INVESTIGACIÓN PRINCIPAL INVESTIGATOR PROJECT TITLE Valencia, Alfonso CancerCureAdvisor : An open bioinformatics platform for personalized treatment of cancer ( Ref.: EUIN2015-62887 ) NETWORKS AND SCIENTIFIC MANAGERS-EUROPE / EUROPA REDES Y GESTORES PRINCIPAL INVESTIGATOR PROJECT TITLE Blasco, Maria A. CNIO in Horizon 2020 : support for proposal preparation and project management ( Ref.: EUC2014-51617 ) ANNUAL REPORT 2016 212 FACTS & FIGURES YOUNG RESEARCHERS PROGRAM / PROGRAMA JÓVENES INVESTIGADORES 10 PRINCIPAL INVESTIGATOR PROJECT TITLE Álvarez, Mónica GPGenCan : Functional relevance of Greatwall/PP2A pathway in the maintenance of genomic stability : therapeutic implications in cancer ( Ref.: SAF2014-60442-JIN ) Lecona, Emilio UBQREP : Modulation of DNA Replication by ubiquitination of chromatin proteins ( Ref.: BFU2014-55168-JIN ) SCIENTIFIC INFRASTRUCTURES / INFRAESTRUCTURSA CIENTÍFICO-TECNOLÓGICAS 11 PRINCIPAL INVESTIGATOR PROJECT TITLE Muñoz, Javier Sistema cromatográfico UHLPC acoplado a Espectrómetro de Masas de alta resolución para estudios de proteómica avanzada ( Ref.: CNIO15-EE-2855 ) Pisano, David G. Clúster SMP de Análisis HPC ( Ref.: CNIO15-EE-3845 ) ASTRAZENECA FOUNDATION / FUNDACIÓN ASTRAZENECA PRINCIPAL INVESTIGATOR PROJECT TITLE Olmos, David Cáncer de próstata familiar y esporádico asociado a alteraciones genéticas, germinales y/o somáticas, en genes de la reparación del DNA Quintela, Miguel Ángel Reprogramación inmune en cáncer de mama preexpuesto a antiangiogénicos inductores de apoxia FERO FOUNDATION / FUNDACIÓN FERO PRINCIPAL INVESTIGATOR PROJECT TITLE Peinado, Héctor Liquid biopsy by nanoplasmonic detection of exosomes : predicting response to ( immuno- and radio )-therapy Valiente, Manuel Predictive biomarkers for brain metastasis in small cell lung cancer SPANISH SOCIETY OF MEDICAL ONCOLOGY / SOCIEDAD ESPAÑOLA DE ONCOLOGÍA MÉDICA ( SEOM ) PRINCIPAL INVESTIGATOR PROJECT TITLE Olmos, David Validación de una firma de expresión con utilidad pronóstica en cáncer de próstata resistente a la castración en una cohorte multi-institucional de pacientes tratados con docetaxel ATRESMEDIA CORPORATION / ATRESMEDIA CORPORACIÓN PRINCIPAL INVESTIGATOR PROJECT TITLE Fernández-Capetillo, Óscar Premio Constantes y Vitales en la categoría “ Joven talento en investigación biomédica ” 2015 ( A3M 2015 ) 10. This Programme is cofunded by the European Regional Development Fund ( ERDF )  11. This Programme is cofunded by the European Regional Development Fund ( ERDF )  SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 213 SCIENTIFIC MANAGEMENT | COMPETITIVE FUNDING OLGA TORRES FOUNDATION / FUNDACIÓN OLGA TORRES PRINCIPAL INVESTIGATOR PROJECT TITLE Djouder, Nabil Understanding the role of growth factors and nutrients in inflammatory bowell disease and colon cancer FUNDACIÓN PROYECTO NEUROFIBROMATOSIS PRINCIPAL INVESTIGATOR PROJECT TITLE Peinado, Héctor Uso de exosomas circulantes como marcadores de progresión en neurofibromatosis y para la determinación de nuevas estrategias terapéuticas BBVA FOUNDATION / FUNDACIÓN BBVA PRINCIPAL INVESTIGATOR PROJECT TITLE Valencia, Alfonso PerMed : Precision Medicine from Big Data to Cognitive Computing CNIO ( Ref.: 76/2016 ) FUNDACIÓN PFIZER PRINCIPAL INVESTIGATOR PROJECT TITLE Peinado, Hector Tumour exosome integrins determine organotropic metastasis ANNUAL REPORT 2016 214 FACTS & FIGURES EDUCATION AND TRAINING PROGRAMMES One of the principal goals of the CNIO is to increase its training capacity in order to give students and professionals the opportunity to advance their careers in the healthcare sector. During 2016, the CNIO signed new agreements with several Spanish Universities and other institutions, namely with the Universidad Autónoma de Barcelona, UNED, the International School of Protocol, University Claude Bernard Lyon, Universidad Francisco de Vitoria, Universidad de Navarra, Universidad de Córdoba, Universidad CEU San Pablo, Fundación Jesús Serra, Fundación ”la Caixa”, and the ISFPS Claudio Galeno of Madrid. TRAINING PROGRAMMES PARTICIPANTS IN EDUCATION AND TRAINING PROGRAMMES 2012 2013 2014 2015 2016 Training of PhD students 121 116 108 105 110 Post-doctoral training 81 67 55 48 51 Training for MDs 16 21 14 25 17 Laboratory training for MSc/BSc students 42 36 73 80 95 Laboratory training for technicians 26 19 21 27 26 Master’s Degree in Molecular Oncology ( graduated ) 37 37 34 29 25 TRAINING OF BSC/MSC STUDENTS The CNIO is committed to training junior scientists at the onset of their careers. To this end, the Centre has established a programme that offers BSc and MSc students the opportunity to obtain hands-on practical laboratory experience by working on ongoing research projects in one of the CNIO groups. The CNIO offers 2 types of short-term laboratory training : ɗ An annual Summer Training Programme for undergraduate students, from any country, who are in their last years of study in the biomedical field. The Programme encompasses 8 weeks of full-time laboratory training ( 285 hours ). During this time, the students actively participate in research projects in one of the CNIO groups. During 2016, 6 students from 3 different countries participated in this programme. ɗ Additionally, students can apply for laboratory training throughout the academic year by directly contacting the Heads of CNIO individual Research Groups or Units. This year, 95 students participated in these programmes, of which 6 ended up joining the CNIO as pre-doctoral students. TRAINING OF PHD STUDENTS The training of PhD students in cutting-edge cancer research is of key importance to the CNIO. The Centre offers many opportunities for bright and dynamic university graduates, of all nationalities, to pursue an ambitious PhD project. To attest this, in 2016, 10 students obtained their PhD degrees and 25 more joined the CNIO. More than 25% of the 110 students working at the CNIO in 2016 were graduates from foreign universities, thus contributing to the internationalisation of the Centre. Since 2008, the Fundación ”la Caixa” offers international fellowships to PhD students to enable them to carry out their thesis projects in biomedical research in Spanish centres of excellence. The CNIO was chosen, as one of 4 such centres, to launch a programme for outstanding young pre-doctoral students from all over the world who have an interest in pursuing an ambitious PhD project. Since 2013, the Ministry of Economy, Industry and Competitiveness has undertaken efforts to link the ”la Caixa”/CNIO International PhD Programme to distinguished research centres accredited as “ Severo Ochoa Centres of Excellence ”. The third call of this new ”la Caixa”- Severo Ochoa International PhD Programme was very successful, attracting around 130 eligible applications from undergraduates from 32 different countries. During 2016, 2 pre-doctoral students received one of these internationally recognised fellowships. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 215 SCIENTIFIC MANAGEMENT | EDUCATION AND TRAINING PROGRAMMES The distribution of students across the CNIO’s Research Programmes in 2016 was as follows : 54.5% of the students worked in the Molecular Oncology Programme, 11.8% in the Cancer Cell Biology Programme, 8.2% in the Structural Biology and Biocomputing Programme, 13.6% in the Human Cancer Genetics Programme, 9.1% in the Clinical Research Programme, 1.8% in the Biotechnology Programme and the remaining 0.9% in the Experimental Therapeutics Programme. FUNDING OF PHD TRAINING NO. SPANISH ENTITIES 86 Ministry of Economy, Industry and Competitiveness / Ministerio de Economía, Industria y Competitividad (MEIC) ( Predoctoral fellowships ) 36 Ministry of Economy, Industry and Competitiveness / Ministerio de Economía, Industria y Competitividad (MEIC) ( I+D Projects ) 7 Ministry of Education, Culture and Sport / Ministerio de Educación, Cultura y Deporte (MECD) ( Predoctoral fellowships ) 4 Institute of Health Carlos III / Instituto de Salud Carlos III (ISCIII) ( I+D Projects ) 6 ”la Caixa” Banking Foundation / Fundación Bancaria ”la Caixa” ( Predoctoral fellowships ) 23 Spanish Association Against Cancer ( AECC ) / Fundación Científica de la AECC ( I+D Projects ) 1 ATresMedia Foundation / Fundación ATresMedia 1 Cris Foundation / Fundación Cris 4 CIBERER 2 CNIO 2 INTERNATIONAL ENTITIES 24 European Commission Framework Programme / H2020 2 Marie Skłodowska-Curie actions of the European Commission 4 European Research Council 5 Portuguese Foundation for Science and Technology (FCT) 1 China Scholarship Council 1 European Foundation for the Study of Diabetes 1 Melanoma Research Alliance 2 Boehringer Ingelheim Fonds 1 Boehringer Ingelheim International GMBH 1 Hoffmann-La Roche 1 Pfizer 3 Prostate Cancer Foundation Young Investigator Award 1 Volkswagen Foundation 1 TOTAL 110 ANNUAL REPORT 2016 216 FACTS & FIGURES POST-DOCTORAL TRAINING One of the CNIO’s prime objectives is to attract young researchers who have recently obtained their PhD or MD degrees and to offer them highly interesting research projects at the forefront of cancer research. In 2016, 51 postdoctoral fellows worked at the CNIO. Notably, more than one third of these fellows were from outside of Spain, many coming from very prestigious international institutions. For yet another year, the Fundación Banco Santander upheld its agreement with the CNIO in 2016 and we continued the highly competitive fellowship programme aimed to support outstanding young scientists who have been trained in the UK or in the USA, and who wish to start or continue their postdoctoral training at the CNIO. One young scientist, who came from the Sloan Kettering Institute for Cancer Research ( New York ), joined the CNIO thanks to a Santander Foundation-CNIO Fellowship in 2016. Another scientist, coming from the City University of New York, was awarded one of these fellowships in late 2016. Thanks to the donations received through the “ CNIO Friends ” platform we launched the inaugural Postdoctoral Contract “ CNIO Friends ” Programme in 2016, thereby enabling us to recruit 2 scientists for a period of 2 years each. Furthermore, in 2016, as a result of an agreement signed with the Juegaterapia Foundation, we were able to create a third Postdoctoral Contract, “ Juegaterapia-CNIO Friends ”, which enabled us to contract a scientist to carry out a project on paediatric oncology. FUNDING SOURCES OF POST-DOCTORAL RESEARCHERS NO. SPANISH ENTITIES 31 Ministry of Economy, Industry and Competitiveness / Ministerio de Economía, Industria y Competitividad (MEIC) ( Postdoctoral fellowships ) 5 Ministry of Economy, Industry and Competitiveness / Ministerio de Economía, Industria y Competitividad (MEIC) ( I+D Projects ) 4 Institute of Health Carlos III / Instituto de Salud Carlos III (ISCIII) ( I+D Projects ) 2 Institute of Health Carlos III / Instituto de Salud Carlos III (ISCIII) ( Postdoctoral fellowship ) 1 Madrid-MIT M+Visión ( Postdoctoral fellowship ) 1 Spanish Association Against Cancer ( AECC ) / Fundación Científica de la AECC ( Postdoctoral fellowships ) 4 Botín Foundation / Fundación Botín 1 Banco Santander Foundation / Fundación Banco Santander 1 Cris Foundation / Fundación Cris 2 La Marató TV3 Foundation / Fundació La Marató TV3 2 HM Hospitals / HM Hospitales 1 CIBERER 1 CNIO 6 INTERNATIONAL ENTITIES 20 European Commission Framework Programme / H2020 2 European Research Council 2 Association for International Cancer Research 5 Melanoma Research Alliance 1 Worldwide Cancer Research UK 1 US Department of Defense 1 Fulbright Commission 1 Boehringer Ingelheim International GMBH 2 Celgene 1 Daiichi Sankyo Agreement 1 Pfizer 3 TOTAL 51 SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 217 SCIENTIFIC MANAGEMENT | EDUCATION AND TRAINING PROGRAMMES POSTGRADUATE PROGRAMMES In addition, the CNIO — in collaboration with several academic institutions in Spain — provides access to a variety of postgraduate programmes that cover the areas of Cellular & Molecular Biology, Molecular Biomedicine, Biotechnology, Biocomputing & Computational Biology, Clinical & Applied Cancer Research, Therapeutic Targets, and Molecular Oncology. Official Postgraduate Programmes in Biosciences The majority of the international postgraduate trainings offered at the CNIO are developed in collaboration with the Faculty of Medicine and Faculty of Sciences at the Autonomous University of Madrid ( UAM ) through 4 Official Postgraduate Programmes, namely the Doctorate in Biosciences, Masters in Molecular and Cell Biology, Masters in Molecular Biomedicine, and Masters in Biotechnology. Master’s Degree in Biocomputing and Computational Biology The Master in Bioinformática y Biología Computacional — directed by Alfonso Valencia, Director of CNIO ’s Structural Biology and Biocomputing Programme — is organised together with the National School of Health of the National Institute of Health Carlos III ( Escuela Nacional de Sanidad del Instituto de Salud Carlos III, ENS-ISCiii ), and the Madrid Science Park ( Parque Científico de Madrid, PCM ). Official Master’s Degree in Clinical and Applied Cancer Research CNIO and the CEU-San Pablo University in Madrid ( USP- CEU ) jointly organise a Postgraduate Training Programme in Clinical and Applied Cancer Research : the Máster Universitario en Investigación Clínica y Aplicada en Oncología. ANNUAL REPORT 2016 218 FACTS & FIGURES Official Master’s Degree in Therapeutic Targets, Research and Development The CNIO collaborates with the Biochemistry and Molecular Biology Department at the University of Alcalá de Henares ( UAH ) for the Máster Oficial en Dianas Terapéuticas, Investigación y Desarrollo. Master’s Degree in Molecular Oncology The main objective of this Master’s degree, organised in collaboration with the Centre for Biomedical Studies ( Centro de Estudios Biosanitarios, CEB ), is to offer training in molecular oncology with emphasis on the latest findings in translational research that are essential for state-of-the art oncological clinical practice. Upon successful completion of the 500 hours of training, a certificate for a Master’s degree in Molecular Oncology — recognised by the European School of Oncology ( ESO ) — is awarded. MÁSTER EN ONCOLOGÍA MOLECULAR Información e inscripciones: Centro de Estudios Biosanitarios e-mail: mefcamara@ceb.edu.es Tlfs. 91 739 52 54 y 630 45 26 62 Información más detallada sobre el programa: www.ceb.edu.es www.cnio.es Junio 2015 - 2017 XI EDICIÓN Bases Moleculares del Cáncer EN COLABORACIÓN CON: Centro Nacional de Investigaciones Oncológicas RECOMENDADO POR: European School of Oncology CON EL AVAL CIENTÍFICO DE: Sociedad Española de Oncología Médica AVALADO POR: Sociedad Española de Hem tología y Hemoterapia SOCIEDAD ESPAÑOLA DE HEMATOLOGÍA Y HEMOTERAPIA SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 219 SCIENTIFIC MANAGEMENT | EDUCATION AND TRAINING PROGRAMMES LABORATORY TRAINING FOR TECHNICIANS This training programme has been developed for students in Anatomical Pathology and is organised through agreements with 18 institutions that provide secondary education for laboratory technicians in Spain. It provides students with hands- on knowledge in cellular and molecular biology techniques. The programme consists of 20 weeks ( 707-712 hours ) of laboratory training for students. Additionally, the CNIO offered real-life work experience to 1 student of Analytical Assays and Quality Control for 12 weeks ( 370 hours ), and to 3 students of Clinical Diagnosis for 12 weeks ( 380 hours ). Of the 26 students who participated in this programme in 2016, 5 were hired by the CNIO. TRAINING FOR MDS In line with CNIO’s commitment to bridge the gap between bench and bedside, the Centre offers 3 programmes providing excellent training opportunities to MDs and other health care professionals. Training usually consists of a 3-month period during residency. In 2016, 17 medical residents from 11 different hospitals enjoyed the benefits of rotations within the different Groups and Units of the CNIO. ADVANCED TRAINING OF SCIENTISTS THROUGH EXTRAMURAL PROGRAMMES During 2016, the Ramón y Cajal Programme supported 8 scientists. This special initiative, established in 2001 by the former Spanish Ministry of Science and Technology (now Spanish Ministry of Economy, Industry and Competitiveness) aims to encourage Spanish or foreign scientists working abroad to return to or relocate to Spain. Successful candidates are selected on the basis of their potential capacity to lead independent projects and groups, or to contribute successfully to the ongoing research in the existing groups. Ten other scientists were funded by similar programmes, including the Juan de la Cierva programme (5 contracts); the ISCIII Miguel Servet (1 contract) and SEOM- Río Hortega (contract funded by the Spanish Society of Medical Oncology, 1 contract) programmes; and the Spanish Association Against Cancer (AECC, 3 contracts). VISITING RESEARCHER PROGRAMME The Jesús Serra Foundation, part of the Catalana Occidente Group, aims to help eminent international specialists work together with CNIO researchers for a few months in order to expand their knowledge in areas of common interest. During 2016, Patrick Sung from the Yale University in New Haven ( USA ) was beneficiary of the Jesús Serra Foundation’s Visiting Researcher Programme. “ SCIENCE BY WOMEN ” PROGRAMME Thanks to this Programme, launched by the Spanish “ Fundación Mujeres por África ”, Dorcas Osei Safo from the University of Ghana, Legon, stayed at the CNIO as a visiting scientist in the Experimental Therapeutics Programme, from January to July 2016. ANNUAL REPORT 2016 220 FACTS & FIGURES SCIENTIFIC EVENTS CNIO-”LA CAIXA” FOUNDATION FRONTIERS MEETINGS The CNIO-”la Caixa ” Foundation Frontiers Meetings are the main international conferences that are jointly organised by the CNIO and the ”la Caixa ” Foundation. They focus on specific, cutting-edge aspects of cancer research, thus providing a unique platform for an intensive and dynamic exchange and debate of scientific ideas. The invited speakers – 20 internationally renowned leaders in oncology – present their latest findings during 2 and a half days. Up to 100 additional participants are selected, via a widely publicised call for applications, based on their potential to make relevant contributions to the conference by presenting hot topics as posters or short talks. CANCEROMATICS III-TUMOR HETEROGENEITY 13-16 NOVEMBER 2016 ORGANISERS · Fátima Al-Shahrour, Spanish National Cancer Research Centre, ( CNIO ), Madrid, Spain · Núria Malats, Spanish National Cancer Research Centre, ( CNIO ), Madrid, Spain · Chris Sander, Dana-Farber Cancer Institute, Boston, US · Alfonso Valencia, Spanish National Cancer Research Centre, ( CNIO ), Madrid, Spain SESSIONS · Pan-Cancer analysis · Analysis of mutations and functional · Consequences of mutations on pathway and network · Tumor heterogeneity · Tumor heterogeneity ( part II ) · Drug prediction and repurposing · Drug prediction and repurposing ( part II ) · Translational Genomics · Translational Genomics ( part II ) SPEAKERS · Bissan Al-Lazikani, The Institute of Cancer Research ( IRC ), London, UK · Niko Beerenwinkel, ETH Zurich, Switzerland · Diego Di Bernardo, Telethon Institute of Genetics and Medicine, Pozzuoli, Italy · Paul Boutros, Ontario Institute for Cancer Research ( OICR ), Toronto, Canada · Fabien Calvo, Cancer Core Europe, Paris, France · Chris Evelo, Maastricht University, The Netherlands SPANISH NATIONAL CANCER RESEARCH CENTRE CNIO - ”LA CAIXA” FOUNDATION FRONTIERS MEETINGS 2016 CANCEROMATICS III — TUMOR HETEROGENEITY Madrid 13 — 16 November 2016 Abstract submission deadline* September 30th Application deadline October 24th Fátima Al-Shahrour Spanish National Cancer Research Centre (CNIO), Madrid, Spain Núria Malats Spanish National Cancer Research Centre (CNIO), Madrid, Spain Alfonso Valencia Spanish National Cancer Research Centre (CNIO), Madrid, Spain Chris Sander Dana-Farber Cancer Institute, Boston, US Confirmed SpeakersOrganisers Bissan Al-Lazikani The Institute of Cancer Research (IRC), London, UK Niko Beerenwinkel ETH Zurich, Switzerland Diego Di Bernardo Telethon Institute of Genetics and Medicine, Pozzuoli, Italy Paul Boutros Ontario Institute for Cancer Research (OICR), Toronto, Canada Fabien Calvo Cancer Core Europe, Paris, France Chris Evelo Maastricht University, The Netherlands Katherine A. Hoadley The UNC Lineberger Comprehensive Cancer Centre, Chapell Hill, US Rachel Karchin Johns Hopkins University (JHU), Baltimore, US Philip Kim University of Toronto, Canada Jan Korbel EMBL, Heidelberg, Germany Hans Lehrach Max Planck Institute for Molecular Genetics, Berlin, Germany Massimo Loda Harvard Medical School, Dana-Farber Cancer Institute, Boston, US Núria López-Bigas University Pompeu Fabra (UPF), Barcelona, Spain Tudor I. Oprea The University of New Mexico (UNM), New Mexico, US Francis Ouellette Ontario Institute for Cancer Research (OICR), Toronto, Canada Jordi Rambla Centre for Genomic Regulation (CRG), Barcelona, Spain Andrea Sottoriva The Institute of Cancer Research, London, UK Charles Swanton The Francis Crick Institute (CRICK), London, UK Andrew E. Teschendorff University College London & CAS-MPG Partner Institute for Computational Biology, London, UK Lodewyk Wessels Netherland Cancer Institute, Amsterdam, Netherland Jessica Zucman-Rossi Inserm, Paris, France For further information and to apply please go to www.cnio.es/events Centro Nacional de Investigaciones Oncológicas Melchor Fernández Almagro 3, 28029 Madrid *Limited capacity of 130 people. CNIO reserves the right to close registration and abstract submission when we reach full capacity. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 221 SCIENTIFIC MANAGEMENT | SCIENTIFIC EVENTS · Katherine A. Hoadley, The UNC Lineberger Comprehensive Cancer Centre, Chapell Hill, US · Rachel Karchin, Johns Hopkins University ( JHU ), Baltimore, US · Philip Kim, University of Toronto, Canada · Jan Korbel, EMBL, Heidelberg, Germany · Hans Lehrach, Max Planck Institute for Molecular Genetics, Berlin, Germany · Massimo Loda, Harvard Medical School, Dana-Farber Cancer Institute, Boston, US · Núria López-Bigas, University Pompeu Fabra ( UPF ), Barcelona, Spain · Tudor I. Oprea, The University of New Mexico, ( UNM ), New Mexico, US · Francis Ouellette, Ontario Institute for Cancer Research ( OICR ), Toronto, Canada · Jordi Rambla, Centre for Genomic Regulation ( CRG ), Barcelona, Spain · Andrea Sottoriva, The Institute of Cancer Research, London, UK · Charles Swanton, The Francis Crick Institute ( CRICK ), London, UK · Andrew E. Teschendorff, University College London & CAS-MPG Partner Institute for Computational Biology, London, UK · Lodewyk Wessels, Netherland Cancer Institute, Amsterdam, Netherland · Jessica Zucman-Rossi, Inserm, Paris, France In addition, 6 short talks were selected among participants ’ contributions and 19 posters were presented. OTHER MEETINGS & CONFERENCES The CNIO annually hosts various international meetings and conferences. Within this category are the 5 international events held in 2016. PANCREOS KICK-OFF MEETING 17 MARCH 2016 Participants by country and supporters of the PancreOS are presented : · Pancreatic Cancer Europe – Alfredo Carrato · EUPancreas COST - Nuria Malats · TTD Spanish Cooperative Group - Inma Ruiz and Juan José García · ENCR/JCR/EU - Carmen Martos BLUEPRINT, ROADMAP, ENCODE AND 1000 GENOMES : BROWSE THROUGH THEM WITH ENSEMBL 4-5 APRIL 2016 ORGANISER · CNIO Bioinformatics SPEAKER · Denise Carvalho-Silva, Ensembl Team, European Bionformatics Institute ( EBI ). ANNUAL REPORT 2016 222 FACTS & FIGURES MAKING ACCESS TO PERSONALISED MEDICINE A REALITY FOR PATIENTS 15 SEPTEMBER 2016 SPEAKERS · Nuria Malats, Spanish National Cancer Research Centre ( CNIO ) · Denis Horgan, EAPM · Antoni Andreu, Ministry of Health · Ramon Gonzalez Carvajal, Regional Ministry of Health of Andalusia · Maria Blasco, Spanish National Cancer Research Centre ( CNIO ) · Natacha Bolaños, Spanish Group for Cancer Patients, GEPAC · Emilia Sanchez Chamorro, Madrid Health Ministry · Ruth Vera, Spanish Society of Medical Oncology ( SEOM ) · Ivo Gut, Centro Nacional de Analisis Genomico CNAG- CRG · Pablo del Pino, Celgene · Federico Plaza, Roche Pharma / Roche Institute VII CONGRESO NACIONAL DE BIOBANCOS Y EL I CONGRESO LATINOAMERICANO DE BIOBANCOS 16-17-18 NOVEMBER 2016 ORGANISERS · Biobanco IDIS · Complejo Hospitalario Universitario de Santiago-CHUS · Fundación Ramón Domínguez · Plataforma Red Nacional de Biobancos · CNIO SPEAKERS · Maimuna Mendy, Agencia Internacional de Investigación en Cáncer ( IARC ). Biobanco IARC · Gustavo Stefanoff, Biobanco del Instituto Nacional de Câncer José Alencar Gomes da Silva ( INCA ) · Balwir Matharoo-Ball, Nottingham University Hospital NHS Trust · Gonzalo Héctor Ardao, Hospital Central de las Fuerzas Armadas ( HCFFAA ) · Hugo Campos, Biobanco del A. C. Camargo Cancer Center · Luz María Ruíz Godoy, Biobanco del Instituto Nacional de Cancerología ( INCan ) · Liliana Virginia Siede, Ministerio de Ciencia, Técnica e Innovación Productiva de la Nación Argentina · Pilar Nicolás, Universidad de Deusto, Bilbao · Máximo Fraga, Biobanco CHUS/IDIS · Jorge Pombo Otero, Servicio de Anatomía Patológica CHUAC · Rocío Aguilar, Biobanco del Sistema Sanitario Público de Andalucía · Ana Caroline Neuber, Biobank, Barretos Cancer Hospital, Barretos, SP · Cristina Villena Portella, Centro Investigación Biomédica en Red, Enfermedades Respiratorias ( CIBERES ) · Diego Santos, Proyecto Coppadis · Susana Teijeira, Banco de Cerebros. Biobanco Vigo. ( IISGS ) I CongresoLatinoamericanode Biobancos Noviembre de 2016 Organiza: Colabora: SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 223 SCIENTIFIC MANAGEMENT | SCIENTIFIC EVENTS · Silvia Sánchez, Instituto de Investigación Sanitaria Hospital La Fe · Gema Huesa, BarcelonaBeta Brain Research Center, Fundación Pasqual Maragall · Manuel Rodríguez Castro, AGAELA · Carmen López Rodríguez, FEGEREC · Anna Bosch-Comas, Biobanc HCB-IDIBAPS · Ángela González Ferro, Hospital Universitario Lucus Augusti · Nuria Ajenjo, Centro Nacional de Investigaciones Oncológicas, CNIO · Natalia Cal Purriños, Fundación Profesor Novoa Santos - Instituto Investigación Biomédica A Coruña ( INIBIC ) · Roberto Bilbao, Biobanco Vasco · Pilar Nicolás, Universidad de Deusto, Bilbao · Rodrigo Dienstmann, Instituto de Oncología del Hospital Vall d’Hebron · Juan Cruz Cigudosa, Centro Nacional de Investigaciones Oncológicas ( CNIO ) · Abel González, Universitat Pompeu Fabra · Manuel M Morente, Plataforma Red Nacional de Biobancos ( ISCIII ) · Inés Aroca Siendones, Biobanco Sistema Sanitario Público de Andalucia · Verónica Valdivieso Gómez, Biobanco SSPA · Ana Garcia Díaz, Centro de Diagnóstico de Enfermedades Moleculares · Raquel Bermudo, Institut d’Investigacions Biomèdiques August Pi i Sunyer ( IDIBAPS ) · Rosa María Pinto Labajo, Banco Nacional de ADN Carlos III, Instituto de Investigación Biomédica de Salamanca · Mara Ortega Gómez, Biobanco HUP. Instituto de Investigación sanitaria Hospital Universitario de la Princesa · Joan Ramón Gómez Cortijo, Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunidad Valenciana · Raquel Amigo, Hospital Universitari i Politècnic La Fe ( IIS-La Fe ) · Gustavo Stefanoff, Instituto Nacional de Cáncer ( INCA )/ Ministerio de Salud · Carol Aristimuño, Biobanco Vasco : Nodo Hospital Universitario Araba · Joan Amoedo Cibeira, GoodGut SL · Andrés García Montero, Banco Nacional de ADN Carlos III · Máximo Fraga, Biobanco CHUS/IDIS · Teresa Escamez, Biobanco en Red de la Región de Murcia ( BIOBANC-MUR ) SENESCENCE & CANCER 2ND ANNUAL MEETING OF THE SPANISH NETWORK OF CELLULAR SENESCENCE 25 NOVEMBER 2016 GROUPS OF THE SPANISH NETWORK : · Joaquín Arribas ( Vall d’Hebron Institute of Oncology, Barcelona, Spain ) and members from his laboratory · Manuel Collado ( Instituto de Investigación Sanitaria – Complejo Hospitalario Universitario de Santiago de Compostela, Spain ) and members from his laboratory · Bill Keyes, ( previously at the Centre of Genomic Regulation, Barcelona, and currently at the IGBMC, Strasbourg, France ) and members from his laboratory · Ramón Martínez ( Univ. Politécnica de Valencia, Valencia, Spain ) and members from his laboratory · Ignacio Palmero ( Instituto de Investigaciones Biomédicas “ Alberto Sols ”, Madrid, Spain ) and members from his laboratory · Manuel Serrano ( CNIO, Madrid, Spain ) and members from his laboratory · Anxo Vidal ( Universidad de Santiago de Compostela, Spain ) and members from his laboratory EXTERNAL INVITED SPEAKERS : · Juan Carlos Acosta, IGMM, Edinburgh, UK · Marco Demaria, ERIBA, Groningen, The Netherlands · Valery Krizhanovsky, Weizmann Institute, Rehovot, Israel ANNUAL REPORT 2016 224 FACTS & FIGURES TRAINING COURSES AND WORKSHOPS The CNIO is committed to disseminating the results of state- of-the-art cancer research to the wider community, including medical professional and junior scientists, thereby enabling them to stay abreast of recent developments in specialised techniques. This is achieved through training courses and hands- on workshops organised by CNIO scientists and and technologists. FLOW CYTOMETRY SEMINAR SERIES 25-26-27 JANUARY 2016 SPEAKERS/ORGANISERS · Rui Gardner, Head of the Flow Cytometry Lab. Instituto Gulbenkian de Ciência, Oeiras, Portugal · Lola Martinez, Head of the Flow Cytometry Unit, CNIO, Madrid, Spain ADVANCED CELL SORTING COURSE 28-29 JANUARY 2016 SPEAKERS/ORGANISERS · Rui Gardner, Head of the Flow Cytometry Lab. Instituto Gulbenkian de Ciência, Oeiras, Portugal · Lola Martinez, Head of the Flow Cytometry Unit, CNIO, Madrid, Spain FORMACIÓN CONTINUADA PARA LA REALIZACIÓN DE LAS FUNCIONES DEL RD/2013 : TÉCNICAS DE IMAGEN EN INVESTIGACIÓN PRECLÍNICA 25 FEBRUARY 2016 SPEAKER · Francisca Mulero, Molecular Imaging Core Unit, Biotechnology Programme, CNIO WORKSHOP REDIEX EXO-IMAGING CNIO 26-28 JULY 2016 SPEAKERS · Hernando del Portillo · Héctor Peinado · Isabel Guerrero · Diego Megias · Juan Monteagudo · Susana Garcia Silva · Ana Amor · Marta Hergueta · Maria Yañez-Mo · Antonio Marcilla · Francesc E. Borrás · Juan Manuel Falcón-Pérez · Antonio Osuna · Lucía Robado SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 225 SCIENTIFIC MANAGEMENT | SCIENTIFIC EVENTS URO-ONCOLOGICAL PATHOLOGY TUTORIAL : A 3-DAY “ MEET THE EXPERT ” 24-25-26 NOVEMBER 2016 ORGANISERS Núria Malats and Francisco X. Real, CNIO, Madrid, Spain TALLER CEGEN-PRB2 : ESTUDIOS DE ASOCIACION : DISEÑO Y ANÁLISIS DE DATOS 29 NOVEMBER 2016 ORGANISER · CEGEN SPEAKERS · Javier Benítez, Director del Nodo del CeGenISCIII en Madrid. Director del Programa de Genética Humana, Centro Nacional de Investigaciones Oncológicas, CNIO · Anna Gonzalez –Neira, Jefe de Unidad de Genotipado Humano-CeGen-ISCIII, Centro Nacional Investigaciones Oncológicas, CNIO · Guillermo Pita, Unidad de Genotipado Humano-CeGen- ISCIII, Centro Nacional Investigaciones Oncológicas, CNIO · Rosario Alonso, Unidad de Genotipado Humano- CeGen- ISCIII, Centro Nacional Investigaciones Oncológicas, CNIO · Pablo Fernández, Jefe de Área de Epidemiología Ambiental y Cáncer, Centro Nacional de Epidemiología ISCIII, Madrid. CIBER en Epidemideología y Salud Pública -CIBERESP · Ana Osorio, Grupo de Genética Humana, Centro Nacional Investigaciones Oncológicas, CNIO · Agustín Fernández Fernández, Unidad de epigenética del Cáncer, Instituto Universitario de Oncología del Principado de Asturias ( IUOPA ) · Sara Ruiz –Pinto, Unidad de Genotipado Humano-CeGen, Centro Nacional Investigaciones Oncológicas, CNIO · Lola Alonso, Grupo de Epidemiologia Genética, Centro Nacional Investigaciones Oncológicas, CNIO COURSE OF ANIMAL LABORATORY FROM C TO D 12-13-14-15-16 DECEMBER 2016 ORGANISERS · CNIO · Animalaria Formación y Gestión SPEAKERS · Manuel Berdoy, University of Oxford, UK · Ignasi Sahun, PCB, Spain · José M. Orellana, University of Alcala, Spain · Sagrario Ortega, CNIO, Spain · Ignacio Álvarez, UCM, Spain · Marlees Leenars, Radbound UMC, Netherlands · Ángel Naranjo, CNB, Spain · Javier Guillén, AAALAC International, Spain · Pedro Pablo López, CNIO, Spain · Violeta Solis, Glaxo Smitkline, Spain · Alba de Martino, CNIO, Spain · Belén Pintado, CBM, Spain · Francisca Mulero, CNIO, Spain · Isabel Blanco, CNIO, Spain MORE INFORMATION www.cnio.es/events or calvaro@srv.cnio.es Uropathology Course A 3-day “Meet the expert” course 24th - 26th November, 2016 Madrid, Spain Speakers: Organisers: Yves Allory Ferran Algaba David Olmos (Henri Mondor Hospital, France) (Fundació Puigvert & Universitat Autònoma de Barcelona, Spain) (CNIO, Spain) Francisco X Real (CNIO, Spain) Núria Malats (CNIO, Spain) CNIO. Melchor Fernández Almagro, 3 28029 Madrid Designed by freepik.es ANNUAL REPORT 2016 226 FACTS & FIGURES CNIO DISTINGUISHED SEMINARS The purpose of the Distinguished Seminars Series is to invite outstanding and internationally renowned scientists to give a seminar and to meet with researchers at the CNIO. Distinguished Seminars are recurrent events that are open to the general public and are held throughout the year, usually on Fridays at noon in the CNIO Auditorium. Each Distinguished Seminar series includes world-leading scientists who address topics that are of general interest to the CNIO faculty. The purpose of this international seminar series is not limited to bringing outstanding cancer researchers to the CNIO, but also serves to annually invite 3 to 4 opinion leaders from other areas of science, technology, and literature ; the overarching goal is to enable the CNIO to present its know-how as well as its vision on contemporary and future technological, societal and cultural challenges. These “ out-of-the-box ” seminars are sponsored by the “ Fundación Banco Sabadell ”. The breadth of expertise and topics covered creates a multidisciplinary and intellectually challenging environment that goes far beyond the frontiers of cancer research. In total, the CNIO hosted 23 distinguished speakers in 2016. Friday 4 Sep James Hurley University of California, Berkeley, USA SEP—DEC 2015 Friday 11 Sep Roger Williams MRC Laboratory of Molecular Biology, Cambridge, UK Friday 18 Sep Siamon Gordon University of Oxford, South Parks Road, Oxford, UK Friday 25 Sep Megan C. King Yale University, New Haven, USA Friday 2 Oct William C. Hahn Dana-Farber Cancer Institute, Boston, USA Friday 9 Oct Eduard Batlle Institute for Research in Biomedicine (IRB Barcelona), Spain Friday 15 Jan Giulio Draetta Institute for Applied Cancer Science, The University of Texas MD Anderson Cancer Center, Houston, USA Friday 30 Oct Hugues de Thé University Institute for Haematology, Paris, France Friday 20 Nov Lee Zou Harvard Medical School, The Jim & Ann Orr MGH Research Scholar, Boston, USA JAN—JUN 2016 Friday 27 Nov Angel Lanas Arbeola Research Health Institute of Aragon, Zaragoza, Spain Friday 4 Dec Carlos Caldas Cancer Research UK Cambridge Institute University of Cambridge, UK Friday 18 Dec Robert Schwabe Columbia University, NY, USA Friday 5 Feb Sarah Teichmann EMBL-European Bioinformatics Institute & Wellcome Trust Sanger Institute, Cambridge, UK Friday 26 Feb Cory Brayton Johns Hopkins University, School of Medicine, Baltimore, USA Friday 4 Mar Michael Sieweke Center of Immunology Marseille-Luminy, France Friday 11 Mar Nicholas Dyson James and Shirley Curvey MGH Research Scholar, Harvard Medical School, Boston, USA Friday 20 May Mathias Heikenwälder Institute of Virology - Helmholtz Center Munich, Germany Friday 15 Apr Andras Nagy Mount Sinai Hospital Lunenfeld-Tanenbaum Research Institute, Toronto, Canada Friday 22 Apr Herbert Waldmann Max-Planck Institute of Molecular Physiology, Dortmund, Germany Friday 29 Apr Navdeep S. Chandel Northwestern University, Feinberg School of Medicine, Chicago, USA Friday 13 May Anna M. Wu Crump Institute for Molecular Imaging, David Geffen School of Medicine at UCLA, USA Friday 10 Jun Adolfo Ferrando Columbia University Medical Center, NY, USA Monday 27 Jun Diane Simeone University of Michigan Health System, Ann Arbor, USA Out-of-the Box Seminars supported by Organisers www.cnio.es/eventos/seminars Melchor Fernández Almagro 3 28029 Madrid, Spain Friday 12 Feb Romain Quidant ICREA, The Institute of Photonic Sciences, Barcelona, Spain Friday 6 May Andrés Moya University of Valencia, FISABIO, Spain Friday 19 Feb Jos Jonkers Netherlands Cancer Institute, Amsterdam Friday 3 Jun Stephan Herzig Institute for Diabetes and Cancer IDC Helmholtz Center, Munich, Germany Friday 28 Oct Charles Brenner University of Iowa Carver College of Medicine Iowa, US Sep—Dec 2016 Friday 21 Oct Mike Hall BiozentrumUniversity of Basel, Switzerland Friday 11 Nov Stig E. Bojesen Herlev Hospital, University of Copenhagen, Denmark Friday 2 Dec Celeste Simon The Abramson Family Cancer Research Institute, University of Pennsylvania Perelman School of Medicine, Philadelphia, US Friday 16 Dec Hans-Guido Wendel Memorial Sloan Kettering Cancer Center, New York, US Friday 12 May Anne Brunet Stanford University School of Medicine Stanford, US Friday 28 Apr Kari Alitalo Institute of Biomedicine, Biomedicum Helsinki, University of Helsinki, Finland Friday 21 Apr Geneviève Almouzni Institut Curie Research Centre, Paris, France Friday 31 Mar José Luis Sanz Autonomous University of Madrid, Spain Friday 5 May Vera Gorbunova University of Rochester, NY, US Friday 19 May Oscar Marín MRC Centre for Developmental Neurobiology, King’s College London, UK Friday 14 Oct Francisco J. Ayala University of California, Irvine, US Friday 16 Sep Francisco J. Martínez Mojica University of Alicante, Spain Out-of-the Box Seminars supported by www.cnio.es/eventos/seminars Melchor Fernández Almagro 3 28029 Madrid, Spain Jan—Jun 2017 Friday 13 Jan Elaine Fuchs Howard Hughes Medical Institute, The Rockefeller University, NY, US Friday 20 Jan Raul Mostoslavsky Massachusetts General Hospital, Harvard Medical School, Boston, US Friday 27 Jan Benjamin L. Ebert Brigham and Women’s Hospital, Harvard Medical School, Boston, US Friday 17 Feb Nuria Oliver DataPop Alliance, New York, US Friday 10 Mar Tom Kirkwood Institute for Cell and Molecular Biosciences, Newcastle University, UK Friday 17 Mar Reinhard Faessler Max Planck Institute of Biochemistry, Munich, Germany Friday 24 Mar Ioannis Aifantis NYU School of Medicine, US Organisers Friday 7 Apr Jacob Hanna Weizmann Institute of Science, Rehovot, Israel Friday 16 Jun Guillermo Oliver Feinberg Cardiovascular Research Institute, Northwestern University, Chicago, US SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 227 SCIENTIFIC MANAGEMENT | SCIENTIFIC EVENTS DATE SPEAKER ORGANISATION TITLE JANUARY 15/01/2016 Giulio Draetta Institute for Applied Cancer Science, The University of Texas MD Anderson Cancer Center, Houston, USA Integrating functional genomics with drug discovery to overcome resistance to treatment in multiple cancer FEBRUARY 05/02/2016 Sarah Teichmann EMBL- European Bioinformatics Institute & Wellcome Trust Sanger Institute, Cambridge, UK Understanding Cellular Heterogeneity 12/02/2016 Romain Quidant ICREA, ICFO- The Institute of Photonic Sciences, Barcelona, Spain Applications to oncology of light and nanotechnology 19/02/2016 Joseph Jonkers Netherlands Cancer Institute, Amsterdam Genetic dissection of breast cancer development, therapy response and resistance in mouse models 26/02/2016 Cory Brayton Johns Hopkins University School of Medicine, Baltimore, USA Research relevant immune variations in mice MARCH 04/03/2016 Michael Sieweke Center of Immunology Marseille-Luminy, France Beyond stem cells : Dissecting lineage identity and self-renewal 11/03/2016 Nicholas Dyson James and Shirley Curvey MGH Research Scholar, Harvard Medical School, Boston, USA The consequences of Rb inactivation APRIL 15/04/2016 Andras Nagy Mount Sinai Hospital Lunenfeld- Tanenbaum Research Institute, Toronto, Canada Reprogramming Leads to Multiple States of Pluripotency in the Artificial Cell Space 22/04/2016 Herbert Waldmann Max-Planck Institute of Molecular Physiology, Dortmund, Germany Chemical Biological Modulation of Ras-Signaling 29/04/2016 Navdeep S. Chandel Northwestern University, Feinberg School of Medicine, Chicago, USA Mitochondria as signaling organelles MAY 06/05/2016 Andrés Moya University of Valencia, Spanish Evolutionary Biology Society ( SESBE ), Valencia, Spain Man : Nature and Future 13/05/2016 Anna M. Wu Crump Institute for Molecular Imaging, David Geffen School of Medicine at UCLA, Los Angeles, US ImmunoPET : Engineered antibodies for noninvasive imaging of tumors and immune cell subsets 20/05/2016 Mathias Heikenwälder DKFZ - German Cancer Research Center, Heidelberg, Germany How cells of the immune system control development of fatty liver disease, non- alcoholic steatohepatitis and subsequent JUNE 03/06/2016 Stephan Herzig Institute for Diabetes and Cancer IDC Helmholtz Center, Munich, Germany Cancer and Metabolism : A bi-directional connection ANNUAL REPORT 2016 228 FACTS & FIGURES 10/06/2016 Adolfo Ferrando Columbia University Medical Center, New York, USA Oncogenic circuitries and mechanisms of resistance in acute lymphoblastic leukemia 27/06/2016 Diane Simeone University of Michigan Health System, Ann Arbor, USA TRIM29 : an Oncogenic Driver in Human Cancers SEPTEMBER 16/09/2016 Francisco Juan Martinez Mojica University of Alicante, Spain The history behind « the CRISPR craze » OCTOBER 14/10/2016 Francisco J. Ayala University of California, Irvine, US Genetic Engineering and Mankind’s Future 21/10/2016 Mike Hall Biozemtrum, University of Basel, Switzerland mTOR signaling in growth and metabolism 28/10/2016 Charles Brenner Roy J. Carver Chair & Head of Biochemistry, University of Iowa, US Nicotinamide Riboside : From Discovery to Human Translation NOVEMBER 11/11/2016 Stig E. Bojesen Herlev and Gentofte Hosptial, Copenhagen University Hospital Faculty of Health Sciences, University of Copenhagen, Denmark DNA and cancer in the general population DECEMBER 02/12/2016 Celeste Simon Abramson Family Cancer Research Institute, University of Pennsylvania School of Medicine, Philadelphia, USA Metabolic adaptations during tumor progression 16/12/2016 Hans-Guido Wendel Memorial Sloan-Kettering Cancer Center, New York, US Restoring tumor suppression with CAR-T micro-pharmacies AD-HOC SEMINARS In addition to the CNIO Distinguished Seminar Series, the CNIO also hosts numerous ad-hoc seminars throughout the year. A total of 49 ad-hoc seminars were organised by CNIO researchers in 2016. DATE SPEAKER ORGANISATION TITLE JANUARY 08/01/2016 Rosa María Señarís University of Santiago de Compostela, Spain Metabolism of cancer cells : Lessons from astrocytic tumors 11/01/2016 Michele de Palma The Swiss Institute for Experimental Cancer Research ( ISREC ), Lausanne, Suiza microRNA regulation of tumor-associated macrophages and response to immunotherapy FEBRUARY 11/02/2016 Jeremy Graff Biothera Pharmaceutical, Inc., Minneapolis, US Imprime PGG, a Pathogen Associated Molecular Pattern ( PAMP ), triggers a coordinated, anti-cancer immune respons SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 229 SCIENTIFIC MANAGEMENT | SCIENTIFIC EVENTS 12/02/2016 Arianna Bianchi Institute of Genetic Medicine, Newcastle University in Newcastle, UK Development of a novel scaffold-free 3D spheroid model of the cornea 22/02/2016 Eva Gonzalez Suarez Bellvitge Biomedical Research Institute ( PEBC ) -IDIBELL Duran i Reynals Hospital, Barcelona, Spain Therapeutic opportunities for RANK pathway in cancer MARCH 01/03/2016 Chris Thorne Horizon Discovery Ltd., Cambridge, United Kingdom Lessons learned from high throughput CRISPR targeting in human cell lines APRIL 12/04/2016 Marc A. Marti- Renom National Centre for Genomic Analysis ( CNAG ) - Center for Genomic Regulation ( CRG ), Barcelona, Spain Structure determination of genomes and genomic domains by satisfaction of spatial restraints 12/04/2016 Karl-Dimiter Bissig Baylor College of Medicine Center for Cell and Gene Therapy, Houston, US From Cancer to metabolic disease - novel applications of human liver chimeric mice 13/04/2016 Rafael De Cabo National Institute on Aging, Baltimore, US Dietary Interventions for Healthy Aging 26/04/2016 Mattia Pelizzola Center for Genomic Science of the IIT, Milan, Italy Epitranscriptomic and regulatory determinants of transcriptional dynamics JUNE 02/06/2016 Dana Branzei IFOM Foundation - The FIRC Institute of Molecular Oncology Foundation, Milan, Italy DNA replication and recombination : tight connections 02/06/2016 Maaike Pols Scientific Outreach Manager Faculty of 1000, London, UK A new way of writing, discovering and sharing science 08/06/2016 Damien P. Devos CABD ; Pablo de Olavide University - CSIC, Sevilla, Spain Deciphering the evolution of genome locus conformation from 4C data 13/06/2016 Eusebio Manchado Novartis Institute for Biomedical Research, Basel, Switzerland Identifying vulnerabilities in undrugabble cancers using RNAi multiplexed technology 16/06/2016 Maria Soriano Carot Addgene, Cambridge, US Addgene, an easy way to share plasmids 20/06/2016 Sean Post and Miguel Gallardo MD Anderson ( Prof. Department of Leukemia ), Houston, US Developing Personalized Therapies for a High-Risk Patient Population with Acute Myeloid Leukemia 22/06/2016 Marek Wagner University of Bergen, Norway The tumor microenvironment and its contribution to melanoma growth and progression 23/06/2016 Alberto Gandarillas Marqués de Valdecilla Institute- IDIVAL, Santander, Spain A novel differentiation-mitosis checkpoint 28/06/2016 Randy Y.C. Poon Hong Kong University of Science and Technology, Kowloon, Hong Kong Exit Strategies – the many fates of mitosis in cancer cells 30/06/2016 Gabriel Victora Whitehead Institute for Biomedical Research, Cambridge, US Clonal Dinamycs in the Antibody Response JULY 14/07/2016 Pekka Katajisto Institute of Biotechnology, University of Helsinki, Finland Age-selective segregation of organelles by stem-like cells 22/07/2016 Kathleen Meyer Institute of Pharmacology and Toxicology, TU Munich, Munich, Germany Essential role for premature senescence of myofibroblasts in myocardial fibrosis 26/07/2016 Michael T. Heneka University of Bonn, Clinic und Polyclinic for Neurology, Bonn, Germany Innate immune activation : a detrimental connection between inflammation and neurodegeneration ANNUAL REPORT 2016 230 FACTS & FIGURES AUGUST 08/08/2016 Luis Arnes Pérez Biomedical Informatics, Columbia University, New York, US Functional analysis of long non-coding RNAs associated with Pancreatic Ductal Adenocarcinoma 09/08/2016 Mark A. Febbraio Garvan Institute of Medical Research, Sidney. Australia Molecular mechanisms for the protective effects of physical exercise on cancer incidence SEPTEMBER 02/09/2016 James DeGregori University of Colorado, Aurora, US Connecting cancer to aging : An evolutionary approach using in silico and in vivo modeling 07/09/2016 Alvaro Ingles Prieto Institute of Science and Technology, Klosterneuburg, Austria Optogenetic control of Receptor Tyrosine Kinases with light-oxygen-voltage 13/09/2016 Guillem Paniagua Soriano PhD student at Leiden University, Netherlands Proteasome inhibitor acquired drug- resistance in Multiple Myeloma 14/09/2016 Yann Cormerais Centre Scientifique de Monaco Amino Acids and Cancer : LAT1 a transporter essential for mTORC1 activity and tumor growth 26/09/2016 Ana Janic The Walter and Eliza Hall Institute, Melbourne, Australia Identification of the critical p53 tumour suppression mechanisms in vivo 26/09/2016 Iain Cheeseman Whitehead Institute for Biomedical Research, Cambridge, US Dissecting the Mechanisms of Cell Division using CRISPR/Cas9 OCTOBER 06/10/2016 Ronald Koop PerkinElmer, Waltham, US Advanced In Vivo Optical Imaging : Tomography, Spectral Unmixing and Co-Registration 13/10/2016 Llucia Albertí Servera Basel University, Switzerland Single-cell analysis of early haematopoietic development : multipotenitality or heterogeneity ? 18/10/2016 Masayuki Yamamoto Tohoku University Graduate School of Medicine, Sendai, Japan The Keap1-Nrf2 System and Cancer Development 21/10/2016 Senthil Muthuswamy Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, US Tumor and Normal Pancreas Organoids : A live biobank platform for discovery and translation 27/10/2016 Maria Sibilia Institute for Cancer Research. Medical University of Vienna, Austria Immune regulation in inflammation and cancer NOVEMBER 03/11/2016 Diana Pippig and Magnus Bauer Ludwig Maximilian University, Munich, Germany Probing Protein Structure and Function by means of Atomic Force Microscopy 17/11/2016 Alejandro Sweet- Cordero Stanford University School of Medicine, US Genomic analysis of osteosarcoma patient- derived xenografts defines therapeutic opportunities and metastatic driver 22/11/2016 Arkaitz Carracedo CIC bioGUNE, University of the Basque Country, Derio, Spain Metabolic rewiring in prostate cancer 22/11/2016 Marta Kovatcheva Koff Laboratory, Memorial Sloan- Kettering Cancer Center, NY, US New roles for old proteins : MDM2, ATRX and HRAS drive the transition from quiescence to senescence 28/11/2016 Eli Gilboa Miller School of Medicine, University of Miami, USA Inducing neoantigens in therapeutic and prophylactic cancer immunotherapy 29/11/2016 Sophie Vasseur CRCM. Cancer Research Center of Marseille, France Decrypting the metabolic flexibility of pancreatic adenocarcinoma SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 231 SCIENTIFIC MANAGEMENT | SCIENTIFIC EVENTS DECEMBER 05/12/2016 Graham Robertson Centenary Institute, Camperdown, Australia Skin inflammation, type 2 immunity and the atopic march 05/12/2016 Amalio Telenti Human Longevity Inc., San Diego, US Genomes are just data 12/12/2016 Jorge Moscat Sanford Burnham Prebys Medical Discovery Institute, La Jolla, US Stroma-epithelium signaling crosstalk by p62 in cancer 14/12/2016 Alfredo Caro CIC bioGUNE | Center for Cooperative Research in Biosciences, Derio, Bilbao Anti-cholesterolemic treatment promotes prostate cancer aggressiveness 15/12/2016 Federico Pietrocola Cordeliers Research Center University of Paris Descartes, Paris, France Caloric Restriction Mimetics : mechanisms and impact on anticancer therapy 16/12/2016 Albert Jordan Molecular Biology Institute of Barcelona, Spain Specificities and genomic distribution of somatic human histone H1 subtypes 21/12/2016 Aranzazu Villasante Columbia University, New York City, US Cancer Engineering : A Translation from One Lan guage to Another CNIO-WOMEN IN SCIENCE ( WISE ) SEMINARS These seminars are aimed to raise gender awareness via lectures given by gender experts and/or role models, and also to provide CNIO researchers with an opportunity to expand their networks. 12/01/2016 María Concepción Ferreras You Tube, Head of Partnerships, Southern Europe and Russia Woman and Technology : a positive story 19/01/2016 Margery Resnick Massachusetts Institute of Technology, Cambridge, US and International Institute in Madrid, Spain In Her Own Voice : MIT’s Earliest Women Scientists 23/02/2016 Pilar Garrido IRYCIS. Ramón y Cajal University Hospital, Madrid, Spain Lung cancer in women : a different disease ? 08/03/2016 Edurne Pasaban Mountaineer, Tolosa, Spain EXPEDITION TO SUCCESS : Achieving goals and overcoming difficulties 10/05/2016 María Teresa Fernández de la Vega President of Women for Africa Foundation ( Fundación Mujeres por África ), Former Vicepresident of Spain Recuerdos y olvidos feministas 28/06/2016 Tánia Balló Documentalist and Film Director, Barcelona, Spain Las Sinsombrero, sin ellas la historia no está completa 27/09/2016 Ángeles González-Sinde Scriptwriter, Film Director. Former Spanish Minister of Culture Out of focus : women and film ( Fuera de foco : mujeres y cine ) 07/11/2016 Guadalupe Martín Martín Radiofísica Hospitalaria/Medical Physicist. Servicio de Radiofísica / Medical Physics Service. Fuenlabrada University Hospital, Madrid, Spain Marie Sklodowska-Curie : Medical Phsycs pioneer and inspiration to female scientists 20/12/2016 Christina Rosenvinge Spanish singer-songwriter, actress and producer 30 years of work : my view on the politics and procedures of the Spanish and global music scenes/30 años de trabajo : mi visión en la política y procedimientos en la escena musical Española y global ANNUAL REPORT 2016 232 FACTS & FIGURES SCIENTIFIC DIVULGATION EVENTS RESEARCHERS ’ NIGHT 30 SEPTEMBER 2016 This year, the CNIO participated in the Researchers ’ Night ; an activity aimed at bringing researchers closer to the general public and concerned families in order to give them the opportunity to learn more about what researchers do for society. Each year, more than 300 European cities participate, in parallel, in what is a great night for science. During the activities − promoted by the European Commission and coordinated by the Madrid Regional Government and the madri+d Foundation − a total of 200 people came to the Spanish National Cancer Research Centre ( CNIO ) to attend Researchers ’ Night ( September 30, 2016 ) and to learn about cancer research. The activities, which were entirely organised by voluntary contributions from 30 young researchers, provided guests the opportunity to meet researchers in an interactive and entertaining way. These included hands-on experiments, view of a virtual tour through the facilities thanks to a video project recorded by scientists from CNIO “ CNIO for Kids ”, and a speed dating session with the researchers. European Researchers’ Night ESPAÑA ANDALUCÍA • MADRID www.lanochedelosinvestigadores.es w w w . m a d r i m a s d . o r g septiembre 30 201 6 O R G A N I Z A NF I N A N C I A conocimiento fundación para el D. L. : M -? ?? ?? -2 01 6. B .O .C .M . C O L A B O R A N noche europea séptima M A D R I D investigadoresde los OPEN DOORS DAY : INVESTIGATING TO DISARM CANCER 7-20 NOVEMBER 2016 The CNIO also dedicates considerable efforts to bringing science and society closer together ; one of these endeavours is its collaboration with the madri+d research network for the organisation of the Madrid Science Week ( XVI Semana de la Ciencia, 7-20 November, 2016 ). In 2016, 50 people participated in the guided visit to the Centre’s facilities. noviembre 2016 7al 20 e n u n l u g a r d e l a c i e n c i a semana de la ciencia d e c i m o s e x t a w w w . m a d r i m a s d . o r g M A D R I D patrocinacolaboran organizan conocimiento fundación para el D. L. : M -? ?? ?? -2 01 6. B .O .C .M . Aplicación disponible para Año Torres Quevedo Centenario del transbordador del Niágara sc semana ciencia teléfonos 010 / 012 SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 233 SCIENTIFIC MANAGEMENT | SCIENTIFIC EVENTS MARIE CURIE EXHIBITION NOVEMBER 2016 The CNIO hosted an exhibition entitled “ Marie Sklodowska- Curie. A Pole in Paris ”; the exhibition revisited the personal and professional life of one of the key women of the 20th century. The exhibition opened on 7 November — the birthday of this Polish-French scientist — and could be visited until the 30th of November. This initiative of the CNIO’s Women in Science Office ( WISE ) stands testament to our commitment to promoting and upholding the work of women scientists. The exhibition attracted 296 visitors. ROCAVIVA EVENTOS BCNMOMENTS 14 JULY 2016 During this year, the company bcnmoments organised the “ Leading Program Madrid ”; a programme sponsored by the ”la Caixa” Foundation that awards the 20 highest selectividad test scores within the Community of Madrid. The selected students had the opportunity to get to know different success stories in a broad range of companies and institutions, including the CNIO. During their “ Business Experience ” at the CNIO in July 2016, the students had the chance to visit the labs guided by young scientists. GUIDED VISITS Throughout the year, the CNIO provides tailor-made opportunities to visit its installations and to learn about the essentials of cancer research. During 2016, more than 471 people participated in such guided visits ; most of them were ESO and Bachillerato student groups, but also professionals in the health sector. ANNUAL REPORT 2016 234 FACTS & FIGURES ADMINISTRATION BOARD OF TRUSTEES ȹ Honorary President · Luis de Guindos Jurado Minister of Economy, Industry and Competitiveness Ministro de Economía, Industria y Competitividad ȹ President · Carmen Vela Olmo Secretariat of State for Research, Development and Innovation of the Spanish Ministry of Economy, Industry and Competitiveness Secretaria de Estado de Investigación, Desarrollo e Innovación del Ministerio de Economía, Industria y Competitividad ȹ Vice-President · Jesús Fernández Crespo Director of the National Institute of Health Carlos III Director del Instituto de Salud Carlos III ȹ Appointed Members · José Javier Castrodeza Sanz Secretary General for Health and Consumer Affairs of the Spanish Ministry of Health, Social Services and Equality Secretario General de Sanidad y Consumo del Ministerio de Sanidad, Servicios Sociales e Igualdad · Marina Pilar Villegas Gracia Director of the Spanish State Research Agency of the Ministry of Economy, Industry and Competitiveness Directora de la Agencia Estatal de Investigación del Ministerio de Economía, Industria y Competitividad · Cristina Ysasi-Ysasmendi Pemán Director of the Department of National Affairs of the Cabinet of the Presidency of the Government Directora del Departamento de Asuntos Nacionales del Gabinete de la Presidencia del Gobierno · Margarita Blázquez Herranz Deputy Director General for Networks and Cooperative Research Centres of the National Institute of Health Carlos III Subdirectora General de Redes y Centros de Investigación Cooperativa del Instituto de Salud Carlos III · Luis Gabilondo Pujol Director General of Health of the Health Department of the Government of Navarre Director General de Salud de la Consejería de Salud del Gobierno de Navarra · Carlos Pesquera González Head of Cabinet of the Healthcare Counsellor of the Government of Cantabria Jefe de Gabinete de la Consejera de Sanidad del Gobierno de Cantabria SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 235 ADMINISTRATION | BOARD OF TRUSTEES · Luis Ángel León Mateos Director of Health Research Planning and Promotion of the Galician Health Service ( SERGAS ) Director del Área de Planificación y Promoción de la Investigación Sanitaria del Servicio Gallego de Salud de la Xunta de Galicia ( SERGAS ) · Sandra García Armesto Managing Director of the Aragon Institute of Health Sciences Directora Gerente del Instituto Aragonés de Ciencias de la Salud ȹ Elected Members · Rafael Pardo Avellaneda Director, BBVA Foundation Director de la Fundación BBVA · Jaume Giró Ribas CEO of ” la Caixa ” Banking Foundation Caixa d ’ Estalvis i Pensions de Barcelona Director General de la Fundación Bancaria Caixa d ’ Estalvis i Pensions de Barcelona, ” la Caixa ” · Ignacio Polanco Moreno Chairman, Grupo PRISA Presidente del Grupo PRISA · Caja Madrid Foundation ( until September 2016 ) Fundación Caja Madrid ȹ Secretary · Margarita Blázquez Herranz Deputy Director General for Networks and Cooperative Research Centres of the National Institute of Health Carlos III Subdirectora General de Redes y Centros de Investigación Cooperativa, Instituto de Salud Carlos III ȹ Legal Advisor · Fernando Arenas Escribano Chief State’s Attorney of the Ministry of Health, Social Services and Equality Abogado del Estado Jefe en el Ministerio de Sanidad, Servicios Sociales e Igualdad * In accordance with the Spanish Transparency Legislation ( Spanish Royal Decree 451/2012, of March 5 ), the following information is hereby provided : - At the close of the financial year, the accumulated remuneration received by the Top Management of the Foundation – the CNIO’s Director plus the Managing Director – has amounted to a total of 203,845 Euros. This amount was received as base salary, seniority, and position supplement. - Members of the CNIO Board of Trustees are not remunerated. ANNUAL REPORT 2016 236 FACTS & FIGURES SCIENTIFIC ADVISORY BOARD · Mariann Bienz, PhD, FRS, FMedSci ( Chair ) Joint Divisional Head Division of Protein and Nucleic Acid Chemistry Medical Research Council Laboratory of Molecular Biology Cambridge, United Kingdom · Lauri A. Aaltonen, MD, PhD Academy Professor Director, Genome Scale Biology Research Programme Biomedicum, University of Helsinki Helsinki, Finland · Genevieve Almouzni, PhD Director, Institut Curie Research Centre Head of Nuclear Dynamics & Genome Plasticity Unit Institut Curie, Paris, France · J. Michael Bishop, MD Chancellor Emeritus Director, G.W. Hooper Research Foundation University of California at San Francisco San Francisco, USA · José Costa, MD, FACP Professor of Pathology and of Orthopaedics and Rehabilitation Director, Translational Diagnostics ; Director, Musculoskeletal Tumor Program Yale University School of Medicine New Haven, USA · Sara Courtneidge, PhD, DSc ( hc ) Associate Director for Translational Sciences, Knight Cancer Institute Professor, Department of Cell, Developmental & Cancer Biology and Biomedical Engineering Oregon Health & Science University Portland, USA · John F.X. Diffley, PhD Associate Research Director The Francis Crick Institute London, United Kingdom · Stephen Frye, PhD Director Center for Integrative Chemical Biology and Drug Discovery Fred Eshelman Distinguished Professor The University of North Carolina at Chapel Hill Chapel Hill, USA · Denise Galloway, PhD Associate Division Director, Human Biology Fred Hutchinson Cancer Research Center Research Professor of Microbiology at the University of Washington Seattle, USA · Scott W. Lowe, PhD Chair, Geoffrey Beene Cancer Research Center Chair, Cancer Biology and Genetics Program Memorial Sloan-Kettering Cancer Center New York, USA · Joan Massagué, PhD Director Sloan Kettering Institute Memorial Sloan-Kettering Cancer Center New York, USA · Ángela Nieto, PhD Full Professor and Head of the Developmental Neurobiology Unit Neuroscience Institute of Alicante ( CSIC-UMH ) Alicante, Spain SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 237 ADMINISTRATION | SCIENTIFIC ADVISORY BOARD · Josep Tabernero, MD PhD Director Vall d’Hebron Institute of Oncology ( VHIO ) Head, Medical Oncology Department of Vall d’Hebron University Hospital P. Vall d’Hebron, Barcelona, Spain · Janet M. Thornton, FRS, PhD Director Emeritus and Senior Scientist European Bioinformatics Institute ( EMBL-EBI ) Hinxton, United Kingdom · Karen H. Vousden, PhD, CBE, FRS, FRSE, FMedSci Director The Beatson Institute for Cancer Research Cancer Research UK Glasgow, United Kingdom · Alfred Wittinghofer, PhD Emeritus Group Leader Department of Structural Biology Max Planck Institute for Molecular Physiology Dortmund, Germany · Ada E. Yonath, PhD Director The Helen and Milton A. Kimmelman Center for Biomolecular Structure and Assembly Kimmel Professor of Structural Biology Weizmann Institute of Science Rehovot, Israel ANNUAL REPORT 2016 238 FACTS & FIGURES MANAGEMENT DIRECTOR SECRETARIATE SECRETARIATE ( COMMUNICATION, INNOVATION, SCIENTIFIC MANAGEMENT ) Alcamí, María Jesús Rodríguez, M. Carmen TECHNOLOGY TRANSFER & VALORISATION OFFICE Sanz, Anabel Director Marín, M. Cruz ( Technology Transfer Manager ) Manzano, Rocío * PROJECTS & CONSORTIA EDUCATION & TRAINING PROGRAMMES SCIENTIFIC EVENTS Molina, Juan Ramón Head Del Codo, Almudena * ( since March ) Moro, Mercedes Head SCIENTIFIC PUBLISHING LIBRARY & ARCHIVES Cerdá, Sonia Head López, Victoria Head DIRECTOR’S OFFICE INTERNATIONAL AFFAIRS Peláez, Fernando Pola, Carolina Director ( since May ) Fernández-Capetillo, Óscar Director COMMUNICATION INNOVATION SCIENTIFIC MANAGEMENT Noriega, Nuria Head Pombo, Vanessa (  Communications Officer ) (until April) De Martos, Cristina ( Communications Officer ) (since April) Barthelemy, Isabel Director Blasco, Maria A. Liébanes, M. Dolores  Head Ares, Raquel Merino, Ana Vergés, Leyre (since December) SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 239 ADMINISTRATION | MANAGEMENT MANAGING DIRECTOR SECRETARIATE Ámez, María del Mar Arroyo, Juan SAP Ferrer, Alfonso Head PURCHASING MAINTENANCE HUMAN RESOURCES PREVENTION & BIOSECURITY ECONOMIC MANAGEMENT INFORMATION TECHNOLOGIES AUDIT Pérez, José Lorenzo Head Bardají, Paz Carbonel, David Martín, Francisco ( until December ) Cespón, Constantino Head Salido, M. Isabel  Head Galindo, José Antonio García, Juan J. Fernández, José Luis Head de Miguel, Marcos Echeverría, Rebeca* ( until December ) García-Risco, Silvia Head Moreno, Ana María Head ( since April ) FINANCE & ADMINISTRATION INFRASTRUCTURE MANAGEMENT EXTRAMURAL CLINICAL RESEARCH Fontaneda, Manuela Director de Dios, Luis Javier Director López, Antonio Director ( until December ) Álamo, Pedro Head Baviano, Marta García-Andrade, Javier Novillo, Angélica De Luna, Almudena * Luongo, Victoria Eloina * Garrido, Fernando  Head ( until May ) Rodríguez, M. José Fernández, Rut * Hernando, M. Elena Doyagüez, Laura * Bertol, Narciso * Plan de Empleo Joven ( Youth Employment Plan ) ANNUAL REPORT 2016 240 FACTS & FIGURES CNIO PERSONNEL 2016 GENDER DISTRIBUTION IN SENIOR ACADEMIC AND MANAGEMENT POSITIONS SCIENTIFIC DIRECTION: DIRECTORS, HEADS OF AREA MANAGEMENT: DIRECTORS, HEADS OF AREA HEADS OF UNIT/BIOBANK FEMALE 50% 7 MALE 50% 7 FEMALE 33% 4 MALE 67% 8 FEMALE 53% 10 MALE 47% 9 GROUP LEADERS, HEADS OF CLINICAL RESEARCH UNIT/SECTION FEMALE 32% 10 MALE 68% 21 420 RESEARCH 91% 152 30 33% 43 > 50 9% 160 31-40 34% 464 TOTAL CNIO PERSONNEL 44 ADMINISTRATION 9% AGE DISTRIBUTION 109 41-50 24% 152 MALE 33% 312 FEMALE 67% GENDER DISTRIBUTION SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 241 SCIENTIFIC PERSONNEL 2016 DISTRIBUTION BY PROGRAMMES EXPERIMENTAL THERAPEUTICS 9% 36 CANCER CELL BIOLOGY 10% 40 BIOTECHNOLOGY 11% 47 CLINICAL RESEARCH 18% 75 MOLECULAR ONCOLOGY 30% 127 STRUCTURAL BIOLOGY AND BIOCOMPUTING 10% 44 HUMAN CANCER GENETICS 12% 51 DISTRIBUTION BY PROFESSIONAL CATEGORY TECHNICIANS 36% 151 PRINCIPAL INVESTIGATORS 12% 50 STAFF SCIENTISTS 17% 72 GRADUATE STUDENTS 25% 104 POST-DOCTORAL FELLOWS 10% 43 GENDER DISTRIBUTION BY PROFESSIONAL CATEGORY TECHNICIANS TOTAL SCIENTIFIC PERSONNEL PRINCIPAL INVESTIGATORS FEMALE 74% 111 MALE 26% 40 FEMALE 283 MALE 137 FEMALE 38% 19 MALE 62% 31 STAFF SCIENTISTS FEMALE 71% 51MALE 29% 21 GRADUATE STUDENTS FEMALE 70% 73 MALE 30% 31 POST-DOCTORAL FELLOWS FEMALE 67% 29 MALE 33% 14 TOTAL SCIENTIFIC PERSONNEL 420 ADMINISTRATION | CNIO PERSONNEL 2016 ANNUAL REPORT 2016 242 FACTS & FIGURES TOTAL SCIENTIFIC PERSONNEL 420 TOTAL 100% 211 TECHNICIANS 34% TECHNICIANS 63% 43 52 TOTAL 100% TOTAL 100% 126 83 DISTRIBUTION BY PROFESSIONAL CATEGORY IN: BASIC RESEARCH PRINCIPAL INVESTIGATORS 11% 23 STAFF SCIENTISTS 15% 32 GRADUATE STUDENTS 34% 71 POST-DOCTORAL FELLOWS 14% 29 TECHNICIANS 26% 56 DISTRIBUTION BY PROFESSIONAL CATEGORY IN: INNOVATION DISTRIBUTION BY PROFESSIONAL CATEGORY IN: TRANSLATIONAL RESEARCH PRINCIPAL INVESTIGATORS 10% PRINCIPAL INVESTIGATORS 17% 13 14 STAFF SCIENTISTS 22% STAFF SCIENTISTS 16% 27 13 GRADUATE STUDENTS 24% 30 POST-DOCTORAL FELLOWS 10% POST-DOCTORAL FELLOWS 1% GRADUATE STUDENTS 3% 13 1 3 SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 243 18 OTHER 38% 6 PORTUGAL 13% 13 ITALY 27% 4 GERMANY 8% 3 AUSTRIA 6% 4 FRANCE 8% SCIENTIFIC PERSONNEL: NATIONAL ORIGIN DISTRIBUTION OF SCIENTIFIC PERSONNEL BY NATIONAL ORIGIN FOREIGN SCIENTIFIC PERSONNEL: DISTRIBUTION BY PROFESSIONAL CATEGORY TECHNICIANS 6% 9 PRINCIPAL INVESTIGATORS 12% 6 STAFF SCIENTISTS 22% 16 GRADUATE STUDENTS 24% 25 POST-DOCTORAL FELLOWS 33% 14 70 NON-SPANISH 17% TOTAL SCIENTIFIC PERSONNEL 100% TOTAL SCIENTIFIC PERSONNEL 100% 420 420 350 SPANISH 83% 48 REST OF EUROPE 11,4% 15 AMERICA 3,6% 7 ASIA 1,7% 350 SPANISH 83,3% Total foreign scientific personnel 70 Percent values represent percentages of foreign employees of the total CNIO personnel in each category ADMINISTRATION | CNIO PERSONNEL 2016 ANNUAL REPORT 2016 244 CNIO Friends SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 245 CNIO Friends 247 ‘ CNIO Friends ’ Postdoctoral Contracts 248 Juegaterapia Foundation-‘ CNIO Friends ’ 249 CNIO opens its doors to ‘CNIO Friends’ 250 Benefactor Friends/Sponsor Friends 252 Donations to the CNIO 253 SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 247 CNIO FRIENDS The ‘ CNIO Friends ’ initiative celebrated its first two years of existence at the end of 2016, and it did so by looking towards the future with all the enthusiasm, commitment, and effort it received from its donors. Since its inception, the community of Friends has continuously increased and more and more people decided to renew their commitment, and thereby their loyalty and trust, to the Centre’s scientific research. We closed 2016 with a community of about 800 Friends, in addition to inheritances and legacies from individuals who decided to support the CNIO’s cutting-edge research ; our challenge now is to get our message across more effectively with every announcement we make. In 2016, thanks to our Friends, we launched the first three cancer research postdoctoral contracts after a competitive assessment process based on scientific excellence. The first two were allocated to sponsor the researchers Paulina Gómez, from the Genetic and Molecular Epidemiology Group, and Vera Pancaldi, from the Structural Computational Biology Group. The former will be working on an international project aimed at analysing the relationship between multiple risk factors and pancreatic cancer, one of the cancers with the highest mortality rates today ; the latter will be studying how DNA structures affect tumour aggressiveness, among other factors. Both researchers stressed “ the crucial importance of philanthropy as a driving force in scientific research and their surprise about the difference between the Anglo-Saxon system - where philanthropy is fully integrated - and our country’s system ”. Indeed, philanthropy is a sign of commitment to a cause, as well as a tool for individuals and society in general to support cancer research regardless of the economic circumstances. The third contract came from the Juegaterapia Foundation and their famous ‘ baby pelones ’ toys, by which they underlined their commitment to childhood cancer research. Last June, the CNIO welcomed one of the most acclaimed popular music stars in our country, Alejandro Sanz, to commemorate the collaboration between the Juegaterapia Foundation and our Centre. Thanks to this grant, the CNIO will host a researcher in 2017 in the Epithelial Carcinogenesis Group who, together with the CNIO- Hospital Niño Jesus Clinical Trials Unit, will study neuroblastic tumours − the most common tumour type in the first 2 years of life − and tumours of the central nervous system, the most common solid tumours in children. Businesses also joined our community of Friends in 2016. Indeed, the CNIO signed a collaborative agreement with ASISA Vida in relation to a new product launched by the insurance company that offers specific coverage for gynaecological cancer. Thanks to the insurance company’s commitment to this cause, 5% of the premiums of this product will be donated to the ‘CNIO Friends ’ initiative to fund cancer research. Another agreement was formalised with Grupo CLH, the leading company in Spain for the transportation and storage of oil products. Our CNIO volunteers are another highly valuable asset. Dozens of them came out last November to meet new Friends at the La Vaguada Shopping Centre, where they installed a mini-laboratory to introduce the initiative to the residents and visitors of the popular Barrio del Pilar district in Madrid, where the centre is located. This initiative was one of many examples that helped raise funds for research : there was the event organised by the secondary school IES La Encantá de Rojales, Alicante, consisting of a charity run in May to raise funds for CNIO’s research ; the Edinburgh Marathon for which participants put on their running shoes in name of cancer research ; and the charity tuna music festival at the Madrid Polytechnic University. Thanks to all of you for supporting our work ! The latest cancer survival figures are encouraging – some types of cancer, such as breast cancer, now exceed 80% – however, there is still a long and difficult road ahead. Our Friends are key in the effort of finding new solutions to fight this disease. Our heartfelt appreciation goes out to each and every one of them. CNIO FRIENDS “ Our Friends : a key partner in our mission of conquering cancer.” ANNUAL REPORT 2016 248 CNIO FRIENDS ‘ CNIO FRIENDS ’ POSTDOCTORAL CONTRACTS Paulina Gómez is an expert in Genetic and Molecular Epidemiology. Her research mainly focuses on PanGenEU, a large European study involving six countries that delves into the relationship between multiple risk factors and pancreatic cancer, one of the carcinomas with the highest mortality rate today. These studies are aimed at defining the population with the greatest risk of suffering from pancreatic cancer, and thereby facilitate the early diagnosis of the disease. “ CNIO Friends is crucial for raising public awareness about the importance of research and for fostering the responsibility we all have for actively participating in our future.” Vera Pancaldi is an expert in Computational Biology. Specifically, she focuses on the study of how the DNA structure affects tumour aggressiveness and how it helps us to understand the context and impact of genetic modifications in patients. These studies could be used as a basis for improving tumour diagnosis and for developing personalised therapies in the clinic. “ This financial contribution has enabled me to ensure the continuity of this project without having to worry about looking for new external funding ; it gives me the necessary independence to develop this research.” At the beginning of the year, the CNIO organised the first recruitment programme funded by donations from the ‘ CNIO Friends ’ philanthropic initiative. After a thorough selection process of the candidates, the first two grants were allocated to sponsor the researchers Paulina Gómez, from the Genetic and Molecular Epidemiology Unit, and Vera Pancaldi, from the Structural Computational Biology Group. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 249 CNIO FRIENDS JUEGATERAPIA FOUNDATION-‘ CNIO FRIENDS ’ In July, the Juegaterapia Foundation, which helps children suffering from cancer, signed a collaboration agreement with the ‘ CNIO Friends ’ initiative to fund a 100,000 euro grant that will be dedicated to research projects related to childhood cancer. Alejandro Sanz came to the CNIO as Juegaterapia’s Goodwill Ambassador to commemorate this agreement. The singer visited the facilities and obtained first-hand information on the research being carried out. “ The fight against cancer is a war and I am a soldier ”, he said during his visit to the CNIO. ANNUAL REPORT 2016 250 CNIO FRIENDS CNIO OPENS ITS DOORS TO ‘CNIO FRIENDS’ One of the most exiting events held in 2016 was the ‘ Jornada Amigos del CNIO ’, which took place in mid-May and welcomed our donors/CNIO Friends in order to bring CNIO science closer to them. Our donors spent an afternoon with us and once again demonstrated their great enthusiasm for being part of the important mission of conquering of cancer. SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 251 CNIO FRIENDS Last but not least, we would also like to extend our heartfelt thanks to all the anonymous benefactors who have donated their legacies to support cancer research at the CNIO ( around €361,000 this year ); in doing so they have contributed to society for generations to come. AMIGOS DEL CNIO más INVESTIGACIÓN menos CÁNCER ANNUAL REPORT 2016 252 CNIO FRIENDS ȹ Benefactor Friends · Alfonso Agüera Nieto Santa Ana-Cartagena, Murcia · Álvaro Gil Conejo Mijas, Málaga · Andrés Sánchez Arranz Madrid, Madrid · Asociación Junta Local Casasimarro Cuenca, Cuenca · Encarnación Fernández Pérez Madrid, Madrid · Ferrán Nacher Carull Xativa, Valencia · Francisco Javier Gállego Franco Barbastro, Huesca · Gema Rubio González Madrid, Madrid · IES La Encantá Rojales, Alicante · Iluminada Hernández González Gijón, Asturias · Instituto Preventivo de Galicia Arteixo, A Coruña · Ismael Crespo Martín Cáceres, Cáceres · Javier Cons García Madrid, Madrid · Jesús Miguel Iglesias Retuerto Valladolid, Valladolid · José Luis Catalá López Las Palmas de Gran Canaria, Las Palmas · José Limiñana Valero Alicante, Alicante · José Polo Criado Cáceres, Cáceres · Juan Félix Ortigosa Córdoba Granollers, Barcelona · Luis David Sanz Navarro Madrid, Madrid · M. Begoña Rumbo Arcas Rutas-Vilaboa/Culleredo, A Coruña · María Jesús Amores Molero Jábaga, Cuenca · María Rodríguez López Celada de los Calderones, Cantabria · Miguel Muñoz Martín Alcalá de Henares, Madrid · Nemesio Carro Carro León, León · Paloma Fuentes González Madrid, Madrid · Raúl Bueno Herrera Plasencia, Cáceres · Santiago Crespo Martín Cáceres, Cáceres · Sergio Recio España Madrid, Madrid · Susana Sanz Fraile Mérida, Badajoz BENEFACTOR FRIENDS/SPONSOR FRIENDS ȹ Sponsor Friends · Asisa Vida Seguros S.A.U. Madrid, Madrid · Compañía Logística de Hidrocarburos CLH, S.A. · Freesia Group Salou, Tarragona · Fundación Juegaterapia Madrid, Madrid · María Josefa Azcona Peribañez Madrid, Madrid SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 253 CNIO FRIENDS DONATIONS TO THE CNIO 505,000€ TOTAL CNIO DONATIONS 2016 248,000€ CNIO FRIENDS 2016 144,000€ 2015 100,000€ 2014 4,000€ 784,000€ LEGACIES 2016 361,000€ 2015 400,000€ 2014 23,000€ 100,000€ ATRESMEDIA 2015 100,000€ 44,000€ DONATIONS BEFORE LAUNCH OF CNIO FRIENDS 2014 10,000€ 2013 25,000€ 2012 3,000€ 2011 6,000€ ANNUAL REPORT 2016 254 CREATIVE TEAM AMPARO GARRIDO PHOTOGRAPHY In order to pour the Annual Report into a more creative concept, the CNIO works closely with selected professionals in the artistic and creative sectors who ensure delivery of an end product that is attractive in more ways than one. We extend our thanks to the creative team, the visual artist Amparo Garrido, and the graphic design studio underbau whose invaluable work created the images and design that illustrate this Annual Report. A Madrid-based visual artist working with photography and video, Amparo Garrido has been represented in individual and group shows both in Spain and abroad since 1998. Her work has been honoured in several prestigious competitions. She obtained the first place in the 2001 edition of the ABC Photography Prize, and second place in the 2007 Purificación García Prize. Other honourable mentions include the Pilar Citoler and Ciudad de Palma prizes. Her work can be found in major collections, including the Museo Nacional Centro de Arte Reina Sofia in Madrid, the photographic holdings of the Madrid regional government, the Coca-Cola Foundation, the Es Baluard Museum of Modern and Contemporary Art in Palma de Mallorca, and the ‘ Types and Trends on the Threshold of the 21st Century ’ Alcobendas Collection, among many others. Amparo’s most recent solo exhibitions in Spain were shown at the Sala Robayera de Miengo, Cantabria 2017, Galería Trinta, Santiago de Compostela 2015, and the Museo del Romanticismo, Madrid 2012. CREATIVE TEAM SPANISH NATIONAL CANCER RESEARCH CENTRE, CNIO 255 CREATIVE TEAM UNDERBAU DESIGN Underbau is a design studio that emerged in 2008 from professional designers with 15 years of experience in the field of corporate design, publishing and advertising. From the very beginning, the studio has sought to maintain its primary focus on art and culture, working together with Spanish and international bodies ( Orquesta y Coro Nacionales de España, Instituto Cervantes, La Fábrica and Museo Thyssen-Bornemisza among others ). Underbau’s total-design approach puts the emphasis on coherency. To achieve that, the studio assumes full responsibility for the entire creative process, from the initial concept to the final product. Centro Nacional de Investigaciones Oncológicas (CNIO) Melchor Fernández Almagro, 3 28029 Madrid, Spain www.cnio.es Coordination Sonia Cerdá Edition Sonia Cerdá and An Devriese Direction Isabel Barthelemy Text, data and figures CNIO Faculty and Management Publication database Victoria López Photo shoot Coordination M. Carmen Rodríguez Photography Amparo Garrido Design underbau Typesetting Nicolás García Marque Prepress La Troupe This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reproduction on microfilms or in any other way, and storage in data banks. © Fundación CNIO Carlos III, 2016 National book catalogue number M-7602-2017 ISSN 2529-9514