Person:
Alonso, Javier

First Name

Javier

Last Name

Alonso

Institution

ISCIII

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ISCIII::Instituto de Investigación de Enfermedades Raras (IIER)

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Now showing 1 - 10 of 37

Therapeutic Potential of EWSR1-FLI1 Inactivation by CRISPR/Cas9 in Ewing Sarcoma.
(Multidisciplinary Digital Publishing Institute (MDPI), 2021-07-27) Cervera Mayor, Saint Thomas; Rodriguez-Martin, Carlos; Fernández-Tabanera, Enrique; Melero-Fernández de Mera, Raquel M; Morin, Matias; Fernández-Peñalver, Sergio; Iranzo-Martínez, Maria; Amhih-Cardenas, Jorge; Garcia-Garcia, Laura; Gonzalez-Gonzalez, Laura; Moreno-Pelayo, Miguel Angel; Alonso, Javier; Instituto de Salud Carlos III; Asociación Pablo Ugarte contra el cáncer infantil; Candela Ribera. Asociación contra el sarcoma de Ewing; Fundación la Sonrisa de Alex para la investigación y el tratamiento del sarcoma de Ewing; Centro de Investigación Biomedica en Red - CIBER; Comunidad de Madrid (España); Asociación Todos somos Iván
Ewing sarcoma is an aggressive bone cancer affecting children and young adults. The main molecular hallmark of Ewing sarcoma are chromosomal translocations that produce chimeric oncogenic transcription factors, the most frequent of which is the aberrant transcription factor EWSR1-FLI1. Because this is the principal oncogenic driver of Ewing sarcoma, its inactivation should be the best therapeutic strategy to block tumor growth. In this study, we genetically inactivated EWSR1-FLI1 using CRISPR-Cas9 technology in order to cause permanent gene inactivation. We found that gene editing at the exon 9 of FLI1 was able to block cell proliferation drastically and induce senescence massively in the well-studied Ewing sarcoma cell line A673. In comparison with an extensively used cellular model of EWSR1-FLI1 knockdown (A673/TR/shEF), genetic inactivation was more effective, particularly in its capability to block cell proliferation. In summary, genetic inactivation of EWSR1-FLI1 in A673 Ewing sarcoma cells blocks cell proliferation and induces a senescence phenotype that could be exploited therapeutically. Although efficient and specific in vivo CRISPR-Cas9 editing still presents many challenges today, our data suggest that complete inactivation of EWSR1-FLI1 at the cell level should be considered a therapeutic approach to develop in the future.
The Transcription Factor FEZF1, a Direct Target of EWSR1-FLI1 in Ewing Sarcoma Cells, Regulates the Expression of Neural-Specific Genes
(Multidisciplinary Digital Publishing Institute (MDPI), 2021-11-12) Garcia-Garcia, Laura; Fernández-Tabanera, Enrique; Cervera Mayor, Saint Thomas; Melero-Fernández de Mera, Raquel M; Josa, Santiago; Gonzalez-Gonzalez, Laura; Rodriguez-Martin, Carlos; Grünewald, Thomas G P; Alonso, Javier; Instituto de Salud Carlos III; Asociación Pablo Ugarte contra el cáncer infantil; Candela Ribera. Asociación contra el sarcoma de Ewing; Asociación Todos somos Iván; Centro de Investigación Biomedica en Red - CIBER; Fundación la Sonrisa de Alex para la investigación y el tratamiento del sarcoma de Ewing
Ewing sarcoma is a rare pediatric tumor characterized by chromosomal translocations that give rise to aberrant chimeric transcription factors (e.g., EWSR1-FLI1). EWSR1-FLI1 promotes a specific cellular transcriptional program. Therefore, the study of EWSR1-FLI1 target genes is important to identify critical pathways involved in Ewing sarcoma tumorigenesis. In this work, we focused on the transcription factors regulated by EWSR1-FLI1 in Ewing sarcoma. Transcriptomic analysis of the Ewing sarcoma cell line A673 indicated that one of the genes more strongly upregulated by EWSR1-FLI1 was FEZF1 (FEZ family zinc finger protein 1), a transcriptional repressor involved in neural cell identity. The functional characterization of FEZF1 was performed in three Ewing sarcoma cell lines (A673, SK-N-MC, SK-ES-1) through an shRNA-directed silencing approach. FEZF1 knockdown inhibited clonogenicity and cell proliferation. Finally, the analysis of the FEZF1-dependent expression profile in A673 cells showed several neural genes regulated by FEZF1 and concomitantly regulated by EWSR1-FLI1. In summary, FEZF1 is transcriptionally regulated by EWSR1-FLI1 in Ewing sarcoma cells and is involved in the regulation of neural-specific genes, which could explain the neural-like phenotype observed in several Ewing sarcoma tumors and cell lines.
CD44 In Sarcomas: A Comprehensive Review and Future Perspectives
(Frontiers Media, 2022-06-17) Fernández-Tabanera, Enrique; Melero-Fernandez de Mera, Raquel M.; Alonso, Javier; Instituto de Salud Carlos III; Asociación Pablo Ugarte contra el cáncer infantil; Candela Ribera. Asociación contra el sarcoma de Ewing; Asociación Todos somos Iván; Fundación la Sonrisa de Alex para la investigación y el tratamiento del sarcoma de Ewing; Centro de Investigación Biomédica en Red - CIBERER (Enfermedades Raras)
It is widely accepted that the tumor microenvironment, particularly the extracellular matrix, plays an essential role in the development of tumors through the interaction with specific protein-membrane receptors. One of the most relevant proteins in this context is the transmembrane protein CD44. The role of CD44 in tumor progression, invasion, and metastasis has been well established in many cancers, although a comprehensive review concerning its role in sarcomas has not been published. CD44 is overexpressed in most sarcomas and several in vitro and in vivo experiments have shown a direct effect on tumor progression, dissemination, and drug resistance. Moreover, CD44 has been revealed as a useful marker for prognostic and diagnostic (CD44v6 isoform) in osteosarcoma. Besides, some innovative treatments such as HA-functionalized liposomes therapy have become an excellent CD44-mediated intracellular delivery system for osteosarcoma. Unfortunately, the reduced number of studies deciphering the prognostic/diagnostic value of CD44 in other sarcoma subgroups, neither than osteosarcoma, in addition to the low number of patients involved in those studies, have produced inconclusive results. In this review, we have gone through the information available on the role of CD44 in the development, maintenance, and progression of sarcomas, analyzing their implications at the prognostic, therapeutic, and mechanistic levels. Moreover, we illustrate how research involving the specific role of CD44 in the different sarcoma subgroups could suppose a chance to advance towards a more innovative perspective for novel therapies and future clinical trials.
DNA Methylomes Reveal Biological Networks Involved in Human Eye Development, Functions and Associated Disorders
(Nature Publishing Group, 2017) Berdasco, María; Gómez, Antonio; Rubio, Marcos J; Català-Mora, Jaume; Zanón-Moreno, Vicente; Lopez, Miguel; Hernández, Cristina; Yoshida, Shigeo; Nakama, Takahito; Ishikawa, Keijiro; Ishibashi, Tatsuro; Boubekeur, Amina M; Louhibi, Lotfi; Pujana, Miguel A; Sayols, Sergi; Setien, Fernando; Corella, Dolores; de Torres, Carmen; Parareda, Andreu; Mora, Jaume; Zhao, Ling; Zhang, Kang; Lleonart, Matilde E; Alonso, Javier; Simó, Rafael; Caminal, Josep M; Esteller, Manel; Fundación La Caixa; Federación Española de Enfermedades Raras; Instituto de Salud Carlos III; Government of Catalonia (España)
This work provides a comprehensive CpG methylation landscape of the different layers of the human eye that unveils the gene networks associated with their biological functions and how these are disrupted in common visual disorders. Herein, we firstly determined the role of CpG methylation in the regulation of ocular tissue-specification and described hypermethylation of retinal transcription factors (i.e., PAX6, RAX, SIX6) in a tissue-dependent manner. Second, we have characterized the DNA methylome of visual disorders linked to internal and external environmental factors. Main conclusions allow certifying that crucial pathways related to Wnt-MAPK signaling pathways or neuroinflammation are epigenetically controlled in the fibrotic disorders involved in retinal detachment, but results also reinforced the contribution of neurovascularization (ETS1, HES5, PRDM16) in diabetic retinopathy. Finally, we had studied the methylome in the most frequent intraocular tumors in adults and children (uveal melanoma and retinoblastoma, respectively). We observed that hypermethylation of tumor suppressor genes is a frequent event in ocular tumors, but also unmethylation is associated with tumorogenesis. Interestingly, unmethylation of the proto-oncogen RAB31 was a predictor of metastasis risk in uveal melanoma. Loss of methylation of the oncogenic mir-17-92 cluster was detected in primary tissues but also in blood from patients.
The role of miR-17-92 in the miRegulatory landscape of Ewing sarcoma
(Impact Journals, 2017-02-14) Schwentner, Raphaela; Herrero-Martin, David; Kauer, Maximilian O; Mutz, Cornelia N; Katschnig, Anna M; Sienski, Grzegorz; Alonso, Javier; Aryee, Dave N T; Kovar, Heinrich; FWF Austrian Science Fund; Unión Europea. Comisión Europea. 7 Programa Marco
MicroRNAs serve to fine-tune gene expression and play an important regulatory role in tissue specific gene networks. The identification and validation of miRNA target genes in a tissue still poses a significant problem since the presence of a seed sequence in the 3'UTR of an mRNA and its expression modulation upon ectopic expression of the miRNA do not reliably predict regulation under physiological conditions. The chimeric oncoprotein EWS-FLI1 is the driving pathogenic force in Ewing sarcoma. MiR-17-92, one of the most potent oncogenic miRNAs, was recently reported to be among the top EWS-FLI1 activated miRNAs. Using a combination of AGO2 pull-down experiments by PAR-CLIP (Photoactivatable-Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation) and of RNAseq upon miRNA depletion by ectopic sponge expression, we aimed to identify the targetome of miR-17-92 in Ewing sarcoma. Intersecting both datasets we found an enrichment of PAR-CLIP hits for members of the miR-17-92 cluster in the 3'UTRs of genes up-regulated in response to mir-17-92 specific sponge expression. Strikingly, approximately a quarter of these genes annotate to the TGFB/BMP pathway, the majority mapping downstream of SMAD signaling. Testing for SMAD phosphorylation, we identify quiet but activatable TGFB signaling and cell autonomous activity of the BMP pathway resulting in the activation of the stemness regulatory transcriptional repressors ID1 and ID3. Taken together, our findings shed light on the complex miRegulatory landscape of Ewing Sarcoma pointing miR-17-92 as a key node connected to TGFB/BMP pathway.
Primary intracranial spindle cell sarcoma with rhabdomyosarcoma-like features share a highly distinct methylation profile and DICER1 mutations.
(Springer, 2018) Koelsche, Christian; Mynarek, Martin; Schrimpf, Daniel; Bertero, Luca; Serrano, Jonathan; Sahm, Felix; Reuss, David E; Hou, Yanghao; Baumhoer, Daniel; Vokuhl, Christian; Flucke, Uta; Petersen, Iver; Brück, Wolfgang; Rutkowski, Stefan; Zambrano, Sandro Casavilca; Garcia Leon, Juan Luis; Diaz Coronado, Rosdali Yesenia; Gessler, Manfred; Tirado, Oscar M; Mora, Jaume; Alonso, Javier; Garcia Del Muro, Xavier; Esteller, Manel; Sturm, Dominik; Ecker, Jonas; Milde, Till; Pfister, Stefan M; Korshunov, Andrey; Snuderl, Matija; Mechtersheimer, Gunhild; Schüller, Ulrich; Jones, David T W; von Deimling, Andreas; German Cancer Aid; Friedberg Charitable Foundation; Sohn Conference Foundation; Fördergemeinschaft Kinderkrebs-Zentrum Hamburg; Damp Foundation
Patients with DICER1 predisposition syndrome have an increased risk to develop pleuropulmonary blastoma, cystic nephroma, embryonal rhabdomyosarcoma, and several other rare tumor entities. In this study, we identified 22 primary intracranial sarcomas, including 18 in pediatric patients, with a distinct methylation signature detected by array-based DNA-methylation profiling. In addition, two uterine rhabdomyosarcomas sharing identical features were identified. Gene panel sequencing of the 22 intracranial sarcomas revealed the almost unifying feature of DICER1 hotspot mutations (21/22; 95%) and a high frequency of co-occurring TP53 mutations (12/22; 55%). In addition, 17/22 (77%) sarcomas exhibited alterations in the mitogen-activated protein kinase pathway, most frequently affecting the mutational hotspots of KRAS (8/22; 36%) and mutations or deletions of NF1 (7/22; 32%), followed by mutations of FGFR4 (2/22; 9%), NRAS (2/22; 9%), and amplification of EGFR (1/22; 5%). A germline DICER1 mutation was detected in two of five cases with constitutional DNA available. Notably, none of the patients showed evidence of a cancer-related syndrome at the time of diagnosis. In contrast to the genetic findings, the morphological features of these tumors were less distinctive, although rhabdomyoblasts or rhabdomyoblast-like cells could retrospectively be detected in all cases. The identified combination of genetic events indicates a relationship between the intracranial tumors analyzed and DICER1 predisposition syndrome-associated sarcomas such as embryonal rhabdomyosarcoma or the recently described group of anaplastic sarcomas of the kidney. However, the intracranial tumors in our series were initially interpreted to represent various tumor types, but rhabdomyosarcoma was not among the typical differential diagnoses considered. Given the rarity of intracranial sarcomas, this molecularly clearly defined group comprises a considerable fraction thereof. We therefore propose the designation "spindle cell sarcoma with rhabdomyosarcoma-like features, DICER1 mutant" for this intriguing group.
Five new cases of syndromic intellectual disability due to KAT6A mutations: widening the molecular and clinical spectrum
(BioMed Central (BMC), 2020-02-10) Urreizti, Roser; Lopez-Martin, Estrella; Martinez-Monseny, Antonio; Pujadas, Montse; Castilla-Vallmanya, Laura; Pérez-Jurado, Luis Alberto; Serrano, Mercedes; Natera-de Benito, Daniel; Martinez-Delgado, Beatriz; Posada De la Paz, Manuel; Alonso, Javier; Marin-Reina, Purificación; O'Callaghan, Mar; Grinberg, Daniel; Bermejo-Sanchez, Eva; Balcells, Susanna; Ministerio de Economía y Competitividad (España); Instituto de Salud Carlos III; Government of Catalonia (España)
BACKGROUND: Pathogenic variants of the lysine acetyltransferase 6A or KAT6A gene are associated with a newly identified neurodevelopmental disorder characterized mainly by intellectual disability of variable severity and speech delay, hypotonia, and heart and eye malformations. Although loss of function (LoF) mutations were initially reported as causing this disorder, missense mutations, to date always involving serine residues, have recently been associated with a form of the disorder without cardiac involvement. RESULTS: In this study we present five new patients, four with truncating mutations and one with a missense change and the only one not presenting with cardiac anomalies. The missense change [p.(Gly359Ser)], also predicted to affect splicing by in silico tools, was functionally tested in the patient's lymphocyte RNA revealing a splicing effect for this allele that would lead to a frameshift and premature truncation. CONCLUSIONS: An extensive revision of the clinical features of these five patients revealed high concordance with the 80 cases previously reported, including developmental delay with speech delay, feeding difficulties, hypotonia, a high bulbous nose, and recurrent infections. Other features present in some of these five patients, such as cryptorchidism in males, syndactyly, and trigonocephaly, expand the clinical spectrum of this syndrome.
Frequency of low-level and high-level mosaicism in sporadic retinoblastoma: genotype-phenotype relationships.
(Springer, 2020-01) Rodriguez-Martin, Carlos; Robledo, Cristina; Gomez-Mariano, Gema Maria; Monzon-Fernandez, Sara; Sastre, Ana; Abelairas, Jose; Sábado, Constantino; Martín-Begué, Nieves; Ferreres, Joan Carles; Fernández-Teijeiro, Ana; González-Campora, Ricardo; Rios-Moreno, María José; Zaballos, Ángel; Cuesta de la Plaza, Isabel; Martinez-Delgado, Beatriz; Posada De la Paz, Manuel; Alonso, Javier; Instituto de Salud Carlos III; Asociación Pablo Ugarte contra el cáncer infantil; Fundación la Sonrisa de Alex para la investigación y el tratamiento del sarcoma de Ewing; Ministerio de Economía y Competitividad (España); Centro de Investigación Biomedica en Red - CIBER; Asociación Todos somos Iván
Somatic mutational mosaicism is a common feature of monogenic genetic disorders, particularly in diseases such as retinoblastoma, with high rates of de novo mutations. The detection and quantification of mosaicism is particularly relevant in these diseases, since it has important implications for genetic counseling, patient management, and probably also on disease onset and progression. In order to assess the rate of somatic mosaicism (high- and low-level mosaicism) in sporadic retinoblastoma patients, we analyzed a cohort of 153 patients with sporadic retinoblastoma using ultra deep next-generation sequencing. High-level mosaicism was detected in 14 out of 100 (14%) bilateral patients and in 11 out of 29 (38%) unilateral patients in whom conventional Sanger sequencing identified a pathogenic mutation in blood DNA. In addition, low-level mosaicism was detected in 3 out of 16 (19%) unilateral patients in whom conventional screening was negative in blood DNA. Our results also reveal that mosaicism was associated to delayed retinoblastoma onset particularly in unilateral patients. Finally we compared the level of mosaicism in different tissues to identify the best DNA source to identify mosaicism in retinoblastoma patients. In light of these results we recommended analyzing the mosaic status in all retinoblastoma patients using accurate techniques such as next-generation sequencing, even in those cases in which conventional Sanger sequencing identified a pathogenic mutation in blood DNA. Our results suggest that a significant proportion of those cases are truly mosaics that could have been overlooked. This information should be taking into consideration in the management and genetic counseling of retinoblastoma patients and families.
CIBERER: Spanish national network for research on rare diseases: A highly productive collaborative initiative
(Wiley, 2022-05) Luque, Juan; Mendes, Ingrid; Gómez, Beatriz; Morte, Beatriz; López de Heredia, Miguel; Herreras, Enrique; Corrochano, Virginia; Bueren, Juan; Gallano, Pía; Artuch, Rafael; Fillat, Cristina; Pérez-Jurado, Luis A; Montoliu, Lluis; Carracedo, Ángel; Millán, José M; Webb, Susan M; Palau, Francesc; CIBERER Networ; Lapunzina, Pablo; Alonso, Javier; Alonso-Ferreira, Veronica; Martinez-Delgado, Beatriz; Instituto de Salud Carlos III; Ministerio de Ciencia e Innovación (España)
CIBER (Center for Biomedical Network Research; Centro de Investigación Biomédica En Red) is a public national consortium created in 2006 under the umbrella of the Spanish National Institute of Health Carlos III (ISCIII). This innovative research structure comprises 11 different specific areas dedicated to the main public health priorities in the National Health System. CIBERER, the thematic area of CIBER focused on rare diseases (RDs) currently consists of 75 research groups belonging to universities, research centers, and hospitals of the entire country. CIBERER's mission is to be a center prioritizing and favoring collaboration and cooperation between biomedical and clinical research groups, with special emphasis on the aspects of genetic, molecular, biochemical, and cellular research of RDs. This research is the basis for providing new tools for the diagnosis and therapy of low-prevalence diseases, in line with the International Rare Diseases Research Consortium (IRDiRC) objectives, thus favoring translational research between the scientific environment of the laboratory and the clinical setting of health centers. In this article, we intend to review CIBERER's 15-year journey and summarize the main results obtained in terms of internationalization, scientific production, contributions toward the discovery of new therapies and novel genes associated to diseases, cooperation with patients' associations and many other topics related to RD research.
Efficacy of ATR inhibitors as single agents in Ewing sarcoma
(Impact Journals, 2016-09-13) Nieto-Soler, Maria; Morgado-Palacin, Isabel; Lafarga, Vanesa; Lecona, Emilio; Murga, Matilde; Callen, Elsa; Azorin, Daniel; Alonso, Javier; Lopez-Contreras, Andres J; Nussenzweig, Andre; Fernandez-Capetillo, Oscar; Banco Santander; Ministerio de Economía y Competitividad (España); Fundación La Marató TV3; Howard Hughes Medical Institute; Instituto de Salud Carlos III; Red Temática de Investigación Cooperativa en Cáncer (RTICC) (España)
Ewing sarcomas (ES) are pediatric bone tumors that arise from a driver translocation, most frequently EWS/FLI1. Current ES treatment involves DNA damaging agents, yet the basis for the sensitivity to these therapies remains unknown. Oncogene-induced replication stress (RS) is a known source of endogenous DNA damage in cancer, which is suppressed by ATR and CHK1 kinases. We here show that ES suffer from high endogenous levels of RS, rendering them particularly dependent on the ATR pathway. Accordingly, two independent ATR inhibitors show in vitro toxicity in ES cell lines as well as in vivo efficacy in ES xenografts as single agents. Expression of EWS/FLI1 or EWS/ERG oncogenic translocations sensitizes non-ES cells to ATR inhibitors. Our data shed light onto the sensitivity of ES to genotoxic agents, and identify ATR inhibitors as a potential therapy for Ewing Sarcomas.