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Permanent URI for this collectionhttps://hdl.handle.net/20.500.12105/19607

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Stem cell niche organization in the Drosophila ovary requires the ECM component Perlecan.
(Cell Press, 2021-04-26) Díaz-Torres, Alfonsa; Rosales-Nieves, Alicia E; Pearson, John R; Santa-Cruz Mateos, Carmen; Marín-Menguiano, Miriam; Marshall, Owen J; Brand, Andrea H; González-Reyes, Acaimo; Agencia Estatal de Investigación (España); Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Royal Society Darwin Trust Research Professorship; Wellcome Trust; European Molecular Biology Organization
Stem cells reside in specialized microenvironments or niches that balance stem cell proliferation and differentiation.1,2 The extracellular matrix (ECM) is an essential component of most niches, because it controls niche homeostasis, provides physical support, and conveys extracellular signals.3-11 Basement membranes (BMs) are thin ECM sheets that are constituted mainly by Laminins, Perlecan, Collagen IV, and Entactin/Nidogen and surround epithelia and other tissues.12 Perlecans are secreted proteoglycans that interact with ECM proteins, ligands, receptors, and growth factors such as FGF, PDGF, VEGF, Hedgehog, and Wingless.13-18 Thus, Perlecans have structural and signaling functions through the binding, storage, or sequestering of specific ligands. We have used the Drosophila ovary to assess the importance of Perlecan in the functioning of a stem cell niche. Ovarioles in the adult ovary are enveloped by an ECM sheath and possess a tapered structure at their anterior apex termed the germarium. The anterior tip of the germarium hosts the germline niche, where two to four germline stem cells (GSCs) reside together with a few somatic cells: terminal filament cells (TFCs), cap cells (CpCs), and escort cells (ECs).19 We report that niche architecture in the developing gonad requires trol, that niche cells secrete an isoform-specific Perlecan-rich interstitial matrix, and that DE-cadherin-dependent stem cell-niche adhesion necessitates trol. Hence, we provide evidence to support a structural role for Perlecan in germline niche establishment during larval stages and in the maintenance of a normal pool of stem cells in the adult niche.
Chromatin dynamics through mouse preimplantation development revealed by single molecule localisation microscopy.
(The Company of Biologists, 2022-08-15) Portela, Marta; Jimenez-Carretero, Daniel; Labrador, Veronica; Andreu, Maria Jose; Arza, Elvira; Caiolfa, Valeria R; Manzanares, Miguel; Agencia Estatal de Investigación (España); Comunidad de Madrid (España); Ministerio de Ciencia e Innovación (España); Fundación Ramón Areces; Instituto de Salud Carlos III; Fundación ProCNIC; Ministerio de Ciencia e Innovación. Centro de Excelencia Severo Ochoa (España)
Most studies addressing chromatin behaviour during preimplantation development are based on biochemical assays that lack spatial and cell-specific information, crucial during early development. Here, we describe the changes in chromatin taking place at the transition from totipotency to lineage specification, by using direct stochastical optical reconstruction microscopy (dSTORM) in whole-mount embryos during the first stages of mouse development. Through the study of two post-translational modifications of Histone 3 related to active and repressed chromatin, H3K4me3 and H3K9me3 respectively, we obtained a time-course of chromatin states, showing spatial differences between cell types, related to their differentiation state. This analysis adds a new layer of information to previous biochemical studies and provides novel insight to current models of chromatin organisation during the first stages of development.
The first wave of the COVID-19 epidemic in Spain was associated with early introductions and fast spread of a dominating genetic variant.
(Nature Publishing Group, 2021) López, Mariana G; Chiner-Oms, Álvaro; García de Viedma, Darío; Ruiz-Rodriguez, Paula; Bracho, Maria Alma; Cancino-Muñoz, Irving; D'Auria, Giuseppe; de Marco, Griselda; García-González, Neris; Goig, Galo Adrian; Gómez-Navarro, Inmaculada; Jiménez-Serrano, Santiago; Martinez-Priego, Llúcia; Ruiz-Hueso, Paula; Ruiz-Roldán, Lidia; Torres-Puente, Manuela; Alberola, Juan; Albert, Eliseo; Aranzamendi Zaldumbide, Maitane; Bea-Escudero, María Pilar; Boga, Jose Antonio; Bordoy, Antoni E; Canut-Blasco, Andrés; Carvajal, Ana; Cilla Eguiluz, Gustavo; Cordón Rodríguez, Maria Luz; Costa-Alcalde, José J; de Toro, María; de Toro Peinado, Inmaculada; Del Pozo, Jose Luis; Duchêne, Sebastián; Fernández-Pinero, Jovita; Fuster Escrivá, Begoña; Gimeno Cardona, Concepción; González Galán, Verónica; Gonzalo Jiménez, Nieves; Hernáez Crespo, Silvia; Herranz, Marta; Lepe, José Antonio; López-Causapé, Carla; López-Hontangas, José Luis; Martín, Vicente; Martró, Elisa; Milagro Beamonte, Ana; Montes Ros, Milagrosa; Moreno-Muñoz, Rosario; Navarro, David; Navarro-Marí, José María; Not, Anna; Oliver, Antonio; Palop-Borrás, Begoña; Parra Grande, Mónica; Pedrosa-Corral, Irene; Pérez González, Maria Carmen; Pérez-Lago, Laura; Pérez-Ruiz, Mercedes; Piñeiro Vázquez, Luis; Rabella, Nuria; Rezusta, Antonio; Robles Fonseca, Lorena; Rodríguez-Villodres, Ángel; Sanbonmatsu-Gámez, Sara; Sicilia, Jon; Soriano, Alex; Tirado Balaguer, María Dolores; Torres, Ignacio; Tristancho, Alexander; Marimón, José María; Coscolla, Mireia; González-Candelas, Fernando; Comas, Iñaki; Instituto de Salud Carlos III; Consejo Superior de Investigaciones Científicas (España); Ministerio de Ciencia (España); Unión Europea. Comisión Europea. European Research Council (ERC); Generalitat Valenciana (España)
The coronavirus disease 2019 (COVID-19) pandemic has affected the world radically since 2020. Spain was one of the European countries with the highest incidence during the first wave. As a part of a consortium to monitor and study the evolution of the epidemic, we sequenced 2,170 samples, diagnosed mostly before lockdown measures. Here, we identified at least 500 introductions from multiple international sources and documented the early rise of two dominant Spanish epidemic clades (SECs), probably amplified by superspreading events. Both SECs were related closely to the initial Asian variants of SARS-CoV-2 and spread widely across Spain. We inferred a substantial reduction in the effective reproductive number of both SECs due to public-health interventions (Re < 1), also reflected in the replacement of SECs by a new variant over the summer of 2020. In summary, we reveal a notable difference in the initial genetic makeup of SARS-CoV-2 in Spain compared with other European countries and show evidence to support the effectiveness of lockdown measures in controlling virus spread, even for the most successful genetic variants.
NLRC4 -mediated activation of CD1c+ dendritic cells contributes to perpetuation of synovitis in rheumatoid arthritis.
(American Society for Clinical Investigation (ASCI), 2022-10-04) Delgado-Arévalo, Cristina; Calvet-Mirabent, Marta; Triguero-Martinez, Ana; Vázquez de Luis, Enrique; Benguría-Filippini, Alberto; Largo, Raquel; Calzada-Fraile, Diego; Popova, Olga; Sánchez-Cerrillo, Ildefonso; Tsukalov, Ilya; Moreno-Vellisca, Roberto; de la Fuente, Hortensia; Herrero-Beaumont, Gabriel; Ramiro, Almudena R; Sánchez-Madrid, Francisco; Castañeda, Santos; Dopazo, Ana; González-Álvaro, Isidoro; Martin-Gayo, Enrique; Comunidad de Madrid (España); Ministerio de Economía y Competitividad (España); Fundación La Caixa; Fundación La Marató TV3; Ministerio de Ciencia e Innovación (España); Instituto de Salud Carlos III; Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)
The individual contribution of specific myeloid subsets such as CD1c+ conventional dendritic cells (cDC) to perpetuation of Rheumatoid Arthritis (RA) pathology remains unclear. In addition, the specific innate sensors driving pathogenic activation of CD1c+ cDCs in RA patients and their functional implications have not been characterized. Here, we assessed phenotypical, transcriptional and functional characteristics of CD1c+ and CD141+ cDCs and monocytes from the blood and synovial fluid of RA patients. Increased levels of CCR2 and the IgG receptor CD64 on circulating CD1c+ cDC associated with the presence of this DC subset in the synovial membrane in RA patients. Moreover, synovial CD1c+ cDCs are characterized by increased expression of proinflammatory cytokines and high abilities to induce pathogenic IFNγ+IL-17+ CD4+ T cells in vitro. Finally, we identified the crosstalk between Fcγ Receptors and NLRC4 as a new potential molecular mechanism mediating pathogenic activation, CD64 upregulation and functional specialization of CD1c+ cDCs in response to dsDNA-IgG in RA patients.
Single cell clonal analysis identifies an AID-dependent pathway of plasma cell differentiation.
(EMBO Press, 2022-10-07) Gómez-Escolar, Carmen; Serrano-Navarro, Alvaro; Benguria, Alberto; Dopazo, Ana; Sanchez-Cabo, Fatima; Ramiro, Almudena R; Fundación La Caixa; Ministerio de Ciencia e Innovación. Centro de Excelencia Severo Ochoa (España); Ministerio de Economía, Industria y Competitividad (España); Ministerio de Ciencia e Innovación (España); Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Unión Europea. Comisión Europea. NextGenerationEU; Instituto de Salud Carlos III; Fundación ProCNIC
Germinal centers (GC) are microstructures where B cells that have been activated by antigen can improve the affinity of their B cell receptors and differentiate into memory B cells (MBCs) or antibody-secreting plasma cells. Here, we have addressed the role of activation-induced deaminase (AID), which initiates somatic hypermutation and class switch recombination, in the terminal differentiation of GC B cells. By combining single cell transcriptome and immunoglobulin clonal analysis in a mouse model that traces AID-experienced cells, we have identified a novel subset of late-prePB cells (L-prePB), which shares the strongest clonal relationships with plasmablasts (PBs). Mice lacking AID have various alterations in the size and expression profiles of transcriptional clusters. We find that AID deficiency leads to a reduced proportion of L-prePB cells and severely impairs transitions between the L-prePB and the PB subsets. Thus, AID shapes the differentiation fate of GC B cells by enabling PB generation from a prePB state.
Remote ischaemic preconditioning ameliorates anthracycline-induced cardiotoxicity and preserves mitochondrial integrity.
(Oxford University Press, 2021-04) Galán-Arriola, Carlos; Villena-Gutierrez, Rocio; Higuero-Verdejo, María I; Diaz-Rengifo, Ivan A; Pizarro, Gonzalo; Lopez, Gonzalo Javier; Molina-Iracheta, Antonio de; Pérez-Martínez, Claudia; García, Rodrigo D; Gonzalez-Calle, David; Lobo-Gonzalez, Manuel; Sánchez, Pedro L; Oliver, Eduardo; Cordoba, Raúl; Fuster, Valentin; Sanchez-Gonzalez, Javier; Ibáñez, Borja; Unión Europea. Comisión Europea. H2020; Unión Europea. Comisión Europea. European Research Council (ERC); Instituto de Salud Carlos III; Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Ministerio de Ciencia e Innovación (España); Fundación ProCNIC
Anthracycline-induced cardiotoxicity (AIC) is a serious adverse effect among cancer patients. A central mechanism of AIC is irreversible mitochondrial damage. Despite major efforts, there are currently no effective therapies able to prevent AIC. Forty Large-White pigs were included. In Study 1, 20 pigs were randomized 1:1 to remote ischaemic preconditioning (RIPC, 3 cycles of 5 min leg ischaemia followed by 5 min reperfusion) or no pretreatment. RIPC was performed immediately before each intracoronary doxorubicin injections (0.45 mg/kg) given at Weeks 0, 2, 4, 6, and 8. A group of 10 pigs with no exposure to doxorubicin served as healthy controls. Pigs underwent serial cardiac magnetic resonance (CMR) exams at baseline and at Weeks 6, 8, 12, and 16, being sacrifice after that. In Study 2, 10 new pigs received 3 doxorubicin injections (with/out preceding RIPC) and were sacrificed at week 6. In Study 1, left ventricular ejection fraction (LVEF) depression was blunted animals receiving RIPC before doxorubicin (RIPC-Doxo), which had a significantly higher LVEF at Week 16 than doxorubicin treated pigs that received no pretreatment (Untreated-Doxo) (41.5 ± 9.1% vs. 32.5 ± 8.7%, P = 0.04). It was mainly due to conserved regional contractile function. In Study 2, transmission electron microscopy (TEM) at Week 6 showed fragmented mitochondria with severe morphological abnormalities in Untreated-Doxo pigs, together with upregulation of fission and autophagy proteins. At the end of the 16-week Study 1 protocol, TEM revealed overt mitochondrial fragmentation with structural fragmentation in Untreated-Doxo pigs, whereas interstitial fibrosis was less severe in RIPC+Doxo pigs. In a translatable large-animal model of AIC, RIPC applied immediately before each doxorubicin injection resulted in preserved cardiac contractility with significantly higher long-term LVEF and less cardiac fibrosis. RIPC prevented mitochondrial fragmentation and dysregulated autophagy from AIC early stages. RIPC is a promising intervention for testing in clinical trials in AIC.
Cross-oncopanel study reveals high sensitivity and accuracy with overall analytical performance depending on genomic regions.
(BioMed Central (BMC), 2021-04) Gong, Binsheng; Li, Dan; Kusko, Rebecca; Novoradovskaya, Natalia; Zhang, Yifan; Wang, Shangzi; Pabon-Peña, Carlos; Zhang, Zhihong; Lai, Kevin; Cai, Wanshi; LoCoco, Jennifer S; Lader, Eric; Richmond, Todd A; Mittal, Vinay K; Liu, Liang-Chun; Johann, Donald J; Willey, James C; Bushel, Pierre R; Yu, Ying; Xu, Chang; Chen, Guangchun; Burgess, Daniel; Cawley, Simon; Giorda, Kristina; Haseley, Nathan; Qiu, Fujun; Wilkins, Katherine; Arib, Hanane; Attwooll, Claire; Babson, Kevin; Bao, Longlong; Bao, Wenjun; Lucas, Anne Bergstrom; Best, Hunter; Bhandari, Ambica; Bisgin, Halil; Blackburn, James; Blomquist, Thomas M; Boardman, Lisa; Burgher, Blake; Butler, Daniel J; Chang, Chia-Jung; Chaubey, Alka; Chen, Tao; Chierici, Marco; Chin, Christopher R; Close, Devin; Conroy, Jeffrey; Cooley Coleman, Jessica; Craig, Daniel J; Crawford, Erin; del Pozo, Angela; Deveson, Ira W; Duncan, Daniel; Eterovic, Agda Karina; Fan, Xiaohui; Foox, Jonathan; Furlanello, Cesare; Ghosal, Abhisek; Glenn, Sean; Guan, Meijian; Haag, Christine; Hang, Xinyi; Happe, Scott; Hennigan, Brittany; Hipp, Jennifer; Hong, Huixiao; Horvath, Kyle; Hu, Jianhong; Hung, Li-Yuan; Jarosz, Mirna; Kerkhof, Jennifer; Kipp, Benjamin; Kreil, David Philip; Łabaj, Paweł; Lapunzina, Pablo; Li, Peng; Li, Quan-Zhen; Li, Weihua; Li, Zhiguang; Liang, Yu; Liu, Shaoqing; Liu, Zhichao; Ma, Charles; Marella, Narasimha; Martín-Arenas, Rubén; Megherbi, Dalila B; Meng, Qingchang; Mieczkowski, Piotr A; Morrison, Tom; Muzny, Donna; Ning, Baitang; Parsons, Barbara L; Paweletz, Cloud P; Pirooznia, Mehdi; Qu, Wubin; Raymond, Amelia; Rindler, Paul; Ringler, Rebecca; Sadikovic, Bekim; Scherer, Andreas; Schulze, Egbert; Sebra, Robert; Shaknovich, Rita; Shi, Qiang; Shi, Tieliu; Silla-Castro, Juan Carlos; Smith, Melissa; Solis-Lopez, Mario; Song, Ping; Stetson, Daniel; Strahl, Maya; Stuart, Alan; Supplee, Julianna; Szankasi, Philippe; Tan, Haowen; Tang, Lin-Ya; Tao, Yonghui; Thakkar, Shraddha; Thierry-Mieg, Danielle; Thierry-Mieg, Jean; Thodima, Venkat J; Thomas, David; Tichy, Boris; Tom, Nikola; Vallespin-Garcia, Elena; Verma, Suman; Walker, Kimbley; Wang, Charles; Wang, Junwen; Wang, Yexun; Wen, Zhining; Wirta, Valtteri; Wu, Leihong; Xiao, Chunlin; Xiao, Wenzhong; Xu, Shibei; Yang, Mary; Ying, Jianming; Yip, Shun H; Zhang, Guangliang; Zhang, Sa; Zhao, Meiru; Zheng, Yuanting; Zhou, Xiaoyan; Mason, Christopher E; Mercer, Timothy; Tong, Weida; Shi, Leming; Jones, Wendell; Xu, Joshua; National Institutes of Health (Estados Unidos); NIH - National Institute of Environmental Health Sciences (NIEHS) (Estados Unidos); United States National Library of Medicine; National Natural Science Foundation of China; National Key R&D Project of China; Shanghai Municipal Science and Technology Major Project; National Health and Medical Research Council (Australia); NIH - National Cancer Institute (NCI) (Estados Unidos)
Targeted sequencing using oncopanels requires comprehensive assessments of accuracy and detection sensitivity to ensure analytical validity. By employing reference materials characterized by the U.S. Food and Drug Administration-led SEquence Quality Control project phase2 (SEQC2) effort, we perform a cross-platform multi-lab evaluation of eight Pan-Cancer panels to assess best practices for oncopanel sequencing. All panels demonstrate high sensitivity across targeted high-confidence coding regions and variant types for the variants previously verified to have variant allele frequency (VAF) in the 5-20% range. Sensitivity is reduced by utilizing VAF thresholds due to inherent variability in VAF measurements. Enforcing a VAF threshold for reporting has a positive impact on reducing false positive calls. Importantly, the false positive rate is found to be significantly higher outside the high-confidence coding regions, resulting in lower reproducibility. Thus, region restriction and VAF thresholds lead to low relative technical variability in estimating promising biomarkers and tumor mutational burden. This comprehensive study provides actionable guidelines for oncopanel sequencing and clear evidence that supports a simplified approach to assess the analytical performance of oncopanels. It will facilitate the rapid implementation, validation, and quality control of oncopanels in clinical use.
Bayesian Inference of Gene Expression
(Exon Publications, 2021-03-20) Jimenez-Jimenez, Victor; Marti-Gomez, Carlos; del Pozo, Miguel Angel; Lara-Pezzi, Enrique; Sanchez-Cabo, Fatima; Ministerio de Economía y Competitividad (España); Instituto de Salud Carlos III; Unión Europea. Comisión Europea. H2020; Fundación ProCNIC; Ministerio de Ciencia e Innovación (España)
Omics techniques have changed the way we depict the molecular features of a cell. The integrative and quantitative analysis of omics data raises unprecedented expectations for understanding biological systems on a global scale. However, its inherently noisy nature, together with limited knowledge of potential sources of variation impacting health and disease, require the use of proper mathematical and computational methods for its analysis and integration. Bayesian inference of probabilistic models allows propagation of the uncertainty from the experimental data to our beliefs of the model parameters, allowing us to appropriately answer complex biological questions. In this chapter, we build probabilistic models of gene expression from RNA-seq data and make inference about their parameters using Bayesian methods. We present models of increasing complexity, from the quantification of a single gene expression to differential gene expression for a whole transcriptome, comparing them to the available tools for analysis of gene expression data. We provide Stan scripts that introduce the reader into the implementation of Bayesian statistics for omics data. The rationale that we apply for transcriptomics data may be easily extended to model the particularities of other omics data and to integrate the different regulatory layers.
Exploratory study of the long-term footprint of deep brain stimulation on brain metabolism and neuroplasticity in an animal model of obesity.
(Nature Publishing Group, 2021-03-10) Casquero-Veiga, Marta; Bueno-Fernandez, Clara; Romero-Miguel, Diego; Lamanna-Rama, Nicolas; Nacher, Juan; Desco, Manuel; Soto-Montenegro, Maria Luisa; Ministerio de Ciencia, Innovación y Universidades (España); Instituto de Salud Carlos III; Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Fundación Mapfre; Fundación Alicia Koplowitz; Fundación Tatiana Pérez de Guzmán el Bueno; Comunidad de Madrid (España); Instituto de Investigación Sanitaria Gregorio Marañón; Ministerio de Ciencia e Innovación (España); Ministerio de Economía y Competitividad (España); Generalitat Valenciana (España)
Deep brain stimulation (DBS) is a powerful neurostimulation therapy proposed for the treatment of several neuropsychiatric disorders. However, DBS mechanism of action remains unclear, being its effects on brain dynamics of particular interest. Specifically, DBS reversibility is a major point of debate. Preclinical studies in obesity showed that the stimulation of the lateral hypothalamus (LH) and nucleus accumbens (NAcc), brain centers involved in satiety and reward circuits, are able to modulate the activity of brain structures impaired in this pathology. Nevertheless, the long-term persistence of this modulation after DBS withdrawal was unexplored. Here we examine the in vivo presence of such changes 1 month after LH- and NAcc-DBS, along with differences in synaptic plasticity, following an exploratory approach. Thus, both stimulated and non-stimulated animals with electrodes in the NAcc showed a common pattern of brain metabolism modulation, presumably derived from the electrodes' presence. In contrast, animals stimulated in the LH showed a relative metabolic invariance, and a reduction of neuroplasticity molecules, evidencing long-lasting neural changes. Our findings suggest that the reversibility or persistence of DBS modulation in the long-term depends on the selected DBS target. Therefore, the DBS footprint would be influenced by the stability achieved in the neural network involved during the stimulation.
Effects of Colchicine on Atherosclerotic Plaque Stabilization: a Multimodality Imaging Study in an Animal Model.
(Springer, 2021-02) Cecconi, Alberto; Vilchez-Tschischke, Jean Paul; Mateo, Jesus; Sanchez-Gonzalez, Javier; Espana, Samuel; Fernandez-Jimenez, Rodrigo; Lopez-Melgar, Beatriz; Fernandez-Friera, Leticia; Lopez-Martin, Gonzalo J.; Fuster, Valentin; Ruiz-Cabello, Jesus; Ibáñez, Borja; Sociedad Española de Cardiología; Instituto de Salud Carlos III; Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Fundación BBVA; Ministerio de Ciencia e Innovación (España); Fundación ProCNIC
Colchicine demonstrated clinical benefits in the treatment of stable coronary artery disease. Our aim was to evaluate the effects of colchicine on atherosclerotic plaque stabilization. Atherosclerosis was induced in the abdominal aorta of 20 rabbits with high-cholesterol diet and balloon endothelial denudation. Rabbits were randomized to receive either colchicine or placebo. All animals underwent MRI, 18F-FDG PET/CT, optical coherence tomography (OCT), and histology. Similar progression of atherosclerotic burden was observed in the two groups as relative increase of normalized wall index (NWI). Maximum 18F-FDG standardized uptake value (meanSUVmax) decreased after colchicine treatment, while it increased in the placebo group with a trend toward significance. Animals with higher levels of cholesterol showed significant differences in favor to colchicine group, both as NWI at the end of the protocol and as relative increase in meanSUVmax. Colchicine may stabilize atherosclerotic plaque by reducing inflammatory activity and plaque burden, without altering macrophage infiltration or plaque typology.
Immune synapse instructs epigenomic and transcriptomic functional reprogramming in dendritic cells.
(American Association for the Advancement of Science (AAAS), 2021-02) Alcaraz-Serna, Ana; Bustos-Moran, Eugenio; Fernandez-Delgado, Irene; Calzada-Fraile, Diego; Torralba, Daniel; Marina-Zarate, Ester; Lorenzo-Vivas, Erika; Vazquez, Enrique; Barreto de Alburquerque, Juliana; Ruef, Nora; Gomez, Manuel J; Sanchez-Cabo, Fatima; Dopazo, Ana; Stein, Jens V; Ramiro, Almudena R; Sanchez-Madrid, Francisco; Ministerio de Economía y Competitividad (España); Comunidad de Madrid (España); Fundación Ramón Areces; Fundación BBVA; Fundación La Caixa; Centro de Investigación Biomedica en Red - CIBER; Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Ministerio de Ciencia, Innovación y Universidades (España); Fundación ProCNIC
Understanding the fate of dendritic cells (DCs) after productive immune synapses (postsynaptic DCs) with T cells during antigen presentation has been largely neglected in favor of deciphering the nuances of T cell activation and memory generation. Here, we describe that postsynaptic DCs switch their transcriptomic signature, correlating with epigenomic changes including DNA accessibility and histone methylation. We focus on the chemokine receptor Ccr7 as a proof-of-concept gene that is increased in postsynaptic DCs. Consistent with our epigenomic observations, postsynaptic DCs migrate more efficiently toward CCL19 in vitro and display enhanced homing to draining lymph nodes in vivo. This work describes a previously unknown DC population whose transcriptomics, epigenomics, and migratory capacity change in response to their cognate contact with T cells.
Human influenza A virus causes myocardial and cardiac-specific conduction system infections associated with early inflammation and premature death.
(Oxford University Press, 2021-02) Filgueiras-Rama, David; Vasilijevic, Jasmina; Jalife, Jose; Noujaim, Sami F; Alfonso-Almazan, Jose M.; Nicolas-Avila, Jose A.; Gutierrez, Celia; Zamarreño, Noelia; Hidalgo, Andres; Bernabe, Alejandro; Cop, Christopher Pablo; Ponce-Balbuena, Daniela; Guerrero-Serna, Guadalupe; Calle, Daniel; Desco, Manuel; Ruiz-Cabello, Jesus; Nieto, Amelia; Falcon, Ana; Fundación La Caixa; European Molecular Biology Organization; Ministerio de Ciencia, Innovación y Universidades (España); Centro de Investigación Biomedica en Red - CIBER; Fundación ProCNIC; Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Frankel Cardiovascular Centre, Michigan Medicine (Estados Unidos); NIH - National Heart, Lung, and Blood Institute (NHLBI) (Estados Unidos)
Human influenza A virus (hIAV) infection is associated with important cardiovascular complications, although cardiac infection pathophysiology is poorly understood. We aimed to study the ability of hIAV of different pathogenicity to infect the mouse heart, and establish the relationship between the infective capacity and the associated in vivo, cellular and molecular alterations. We evaluated lung and heart viral titres in mice infected with either one of several hIAV strains inoculated intranasally. 3D reconstructions of infected cardiac tissue were used to identify viral proteins inside mouse cardiomyocytes, Purkinje cells, and cardiac vessels. Viral replication was measured in mouse cultured cardiomyocytes. Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were used to confirm infection and study underlying molecular alterations associated with the in vivo electrophysiological phenotype. Pathogenic and attenuated hIAV strains infected and replicated in cardiomyocytes, Purkinje cells, and hiPSC-CMs. The infection was also present in cardiac endothelial cells. Remarkably, lung viral titres did not statistically correlate with viral titres in the mouse heart. The highly pathogenic human recombinant virus PAmut showed faster replication, higher level of inflammatory cytokines in cardiac tissue and higher viral titres in cardiac HL-1 mouse cells and hiPSC-CMs compared with PB2mut-attenuated virus. Correspondingly, cardiac conduction alterations were especially pronounced in PAmut-infected mice, associated with high mortality rates, compared with PB2mut-infected animals. Consistently, connexin43 and NaV1.5 expression decreased acutely in hiPSC-CMs infected with PAmut virus. YEM1L protease also decreased more rapidly and to lower levels in PAmut-infected hiPSC-CMs compared with PB2mut-infected cells, consistent with mitochondrial dysfunction. Human IAV infection did not increase myocardial fibrosis at 4-day post-infection, although PAmut-infected mice showed an early increase in mRNAs expression of lysyl oxidase. Human IAV can infect the heart and cardiac-specific conduction system, which may contribute to cardiac complications and premature death.
Activation of amino acid metabolic program in cardiac HIF1-alpha-deficient mice.
(Cell Press, 2021-02-19) Menendez-Montes, Ivan; Escobar, Beatriz; Gomez, Manuel J; Albendea-Gomez, Teresa; Palacios, Beatriz; Bonzon-Kulichenko, Elena; Izquierdo-Garcia, Jose Luis; Alonso, Ana Vanessa; Ferrarini, Alessia; Jimenez-Borreguero, Luis J; Ruiz-Cabello, Jesus; Vazquez, Jesus; Martin-Puig, Silvia; Centro Nacional de Investigaciones Cardiovasculares Carlos III (España); Unión Europea. Comisión Europea. European Research Council (ERC); Fundación La Marató TV3; Comunidad de Madrid (España); Instituto de Salud Carlos III; Francisco de Vitoria University (España); Fondation Leducq; Fundación La Caixa
HIF1-alpha expression defines metabolic compartments in the developing heart, promoting glycolytic program in the compact myocardium and mitochondrial enrichment in the trabeculae. Nonetheless, its role in cardiogenesis is debated. To assess the importance of HIF1-alpha during heart development and the influence of glycolysis in ventricular chamber formation, herein we generated conditional knockout models of Hif1a in Nkx2.5 cardiac progenitors and cardiomyocytes. Deletion of Hif1a impairs embryonic glycolysis without influencing cardiomyocyte proliferation and results in increased mitochondrial number and transient activation of amino acid catabolism together with HIF2α and ATF4 upregulation by E12.5. Hif1a mutants display normal fatty acid oxidation program and do not show cardiac dysfunction in the adulthood. Our results demonstrate that cardiac HIF1 signaling and glycolysis are dispensable for mouse heart development and reveal the metabolic flexibility of the embryonic myocardium to consume amino acids, raising the potential use of alternative metabolic substrates as therapeutic interventions during ischemic events.
Todos los caminos llevan a Roma: resultados de las terapias respiratorias no invasivas aplicadas en un centro terciario sin Unidad de Cuidados Intermedios durante la pandemia de COVID-19
(Elsevier, 2021-01) Lopez-Padilla, Daniel; Teran-Tinedo, Jose Rafael; Lopez-Martin, Soledad; Caballero-Segura, Francisco Jose; Gallo-Gonzalez, Virginia; Recio-Moreno, Beatriz; Ji, Zichen; Castro-Riera, Christian; Ojeda-Castillejo, Elena; Desco, Manuel; Benedetti, Paola; Cerezo-Lajas, Alicia; Miguel-Diez, Javier de; Dominguez-Zabaleta, Irene Milagros; Ferreira-Moreno, Alicia; Fuentes-Alonso, Marta; Garcia-de-Pedro, Julia; Garcia-Lopez, Javier; Giron-Matute, Walther Ivan; Gomez-Garcia, Rosa M; Liendo-Martínez, Katiuska; Oliva-Ramos, Alicia; Parra-Leon, Virginia; Pedraza-Serrano, Fernando; Sanchez-Muñoz, Gema; Sanz-Sanz, Pilar; Suarez-Escudero, Sergio; Vargas-Espinal, Julio; Sousa-Casasnovas, Iago; Diez-del Hoyo, Felipe; Puente-Maestu, Luis
Introducción. Las terapias respiratorias no invasivas (TRNI) fueron ampliamente utilizadas en la primera ola de la pandemia de COVID-19, en escenarios distintos según los medios disponibles. El objetivo fue presentar la supervivencia a 90 días y los factores asociados a esta de los pacientes tratados con TRNI en un centro de tercer nivel sin Unidad de Cuidados Respiratorios Intermedios. Como objetivo secundario comparar los resultados obtenidos de las distintas terapias. Métodos. Estudio observacional de pacientes tratados con TRNI fuera de un ambiente de Cuidados Intensivos o Unidad de Cuidados Respiratorios Intermedios, diagnosticados de COVID-19 y con síndrome de distrés respiratorio agudo por criterios radiológicos y de ratio SpO2/FiO2. Se desarrolló un modelo multivariante de regresión logística para determinar las variables independientemente asociadas, y se compararon los resultados de la terapia de alto flujo con cánula nasal y la presión positiva continua en la vía aérea. Resultados. Se trataron 107 pacientes y sobrevivieron 85 (79,4%) a los 90 días. Antes de iniciar la TRNI el ratio medio de SpO2/FiO2 fue de 119,8±59,4. Un mayor score de SOFA se asoció significativamente a la mortalidad (OR 2,09; IC95% 1,34 – 3,27), mientras que la autopronación fue un factor protector (OR 0,23; IC95% 0,06 – 0,91). La terapia de alto flujo con cánula nasal fue utilizada en 63 sujetos (58,9%), y la presión positiva continua en la vía aérea en 41 (38,3%). No se encontraron diferencias entre ellas. Conclusión. Aproximadamente cuatro de cada cinco pacientes tratados con TRNI sobrevivieron a los 90 días, y no se encontraron diferencias significativas entre la terapia de alto flujo con cánula nasal y la presión positiva continua en la vía aérea.
Covalently Labeled Fluorescent Exosomes for In Vitro and In Vivo Applications.
(Multidisciplinary Digital Publishing Institute (MDPI), 2021-01-16) Gonzalez, Maria Isabel; Gonzalez-Arjona, Mario; Santos-Coquillat, Ana; Vaquero, Javier; Vazquez-Ogando, Elena; Molina-Iracheta, Antonio; Peinado Selgas, Hector; Desco, Manuel; Salinas, Beatriz; Comunidad de Madrid (España); Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Instituto de Salud Carlos III; Ministerio de Ciencia e Innovación (España); Fundación ProCNIC
The vertiginous increase in the use of extracellular vesicles and especially exosomes for therapeutic applications highlights the necessity of advanced techniques for gaining a deeper knowledge of their pharmacological properties. Herein, we report a novel chemical approach for the robust attachment of commercial fluorescent dyes to the exosome surface with covalent binding. The applicability of the methodology was tested on milk and cancer cell-derived exosomes (from U87 and B16F10 cancer cells). We demonstrated that fluorescent labeling did not modify the original physicochemical properties of exosomes. We tested this nanoprobe in cell cultures and healthy mice to validate its use for in vitro and in vivo applications. We confirmed that these fluorescently labeled exosomes could be successfully visualized with optical imaging.
The Paternal Transition Entails Neuroanatomic Adaptations that are Associated with the Father's Brain Response to his Infant Cues.
(Oxford University Press, 2020-11) Paternina-Die, Maria; Martinez-Garcia, Magdalena; Pretus, Clara; Hoekzema, Elseline; Barba-Müller, Erika; Martin de Blas, Daniel; Pozzobon, Cristina; Ballesteros, Agustin; Vilarroya, Oscar; Desco, Manuel; Carmona, Susanna; Ministerio de Ciencia, Innovación y Universidades (España); Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Fundación La Caixa; Unión Europea. Comisión Europea. European Research Council (ERC); Fundación ProCNIC; Unión Europea. Comisión Europea. H2020; Instituto de Salud Carlos III
The transition into fatherhood is a life-changing event that requires substantial psychological adaptations. In families that include a father figure, sensitive paternal behavior has been shown to positively impact the infant's development. Yet, studies exploring the neuroanatomic adaptations of men in their transition into fatherhood are scarce. The present study used surface-based methods to reanalyze a previously published prospective magnetic resonance imaging dataset comprised of 20 first-time fathers (preconception-to-postpartum) and 17 childless men. We tested if the transition into fatherhood entailed changes in cortical volume, thickness, and area and whether these changes were related to 2 indicators of paternal experience. Specifically, we tested if such changes were associated with (1) the baby's age and/or (2) the fathers' brain activity in response to pictures of their babies compared with an unknown baby. Results indicated that first-time fathers exhibited a significant reduction in cortical volume and thickness of the precuneus. Moreover, higher volume reduction and cortical thinning were associated with stronger brain responses to pictures of their own baby in parental brain regions. This is the first study showing preconception-to-postpartum neuroanatomical adaptations in first-time fathers associated with the father's brain response to cues of his infant.
Interplay between UNG and AID governs intratumoral heterogeneity in mature B cell lymphoma.
(Public Library of Science (PLOS), 2020-12) Delgado, Pilar; Alvarez-Prado, Angel Francisco; Marina-Zarate, Ester; Sernandez, Isora V.; Mur, Sonia M.; de la Barrera, Jorge; Sanchez-Cabo, Fatima; Cañamero, Marta; Molina-Iracheta, Antonio; Belver, Laura; de Yebenes, Virginia G; Ramiro, Almudena R
Most B cell lymphomas originate from B cells that have germinal center (GC) experience and bear chromosome translocations and numerous point mutations. GC B cells remodel their immunoglobulin (Ig) genes by somatic hypermutation (SHM) and class switch recombination (CSR) in their Ig genes. Activation Induced Deaminase (AID) initiates CSR and SHM by generating U:G mismatches on Ig DNA that can then be processed by Uracyl-N-glycosylase (UNG). AID promotes collateral damage in the form of chromosome translocations and off-target SHM, however, the exact contribution of AID activity to lymphoma generation and progression is not completely understood. Here we show using a conditional knock-in strategy that AID supra-activity alone is not sufficient to generate B cell transformation. In contrast, in the absence of UNG, AID supra-expression increases SHM and promotes lymphoma. Whole exome sequencing revealed that AID heavily contributes to lymphoma SHM, promoting subclonal variability and a wider range of oncogenic variants. Thus, our data provide direct evidence that UNG is a brake to AID-induced intratumoral heterogeneity and evolution of B cell lymphoma.
Single breath-hold saturation recovery 3D cardiac T1 mapping via compressed SENSE at 3T.
(Springer, 2020-12) Ferreira da Silva, Tiago; Galan-Arriola, Carlos; Montesinos, Paula; Lopez-Martin, Gonzalo J.; Desco, Manuel; Fuster, Valentin; Ibáñez, Borja; Sanchez-Gonzalez, Javier; Unión Europea. Comisión Europea. H2020; Instituto de Salud Carlos III; Comunidad de Madrid (España); Ministerio de Ciencia, Innovación y Universidades (España); Fundación ProCNIC
To propose and validate a novel imaging sequence that uses a single breath-hold whole-heart 3D T1 saturation recovery compressed SENSE rapid acquisition (SACORA) at 3T. The proposed sequence combines flexible saturation time sampling, compressed SENSE, and sharing of saturation pulses between two readouts acquired at different RR intervals. The sequence was compared with a 3D saturation recovery single-shot acquisition (SASHA) implementation with phantom and in vivo experiments (pre and post contrast; 7 pigs) and was validated against the reference inversion recovery spin echo (IR-SE) sequence in phantom experiments. Phantom experiments showed that the T1 maps acquired by 3D SACORA and 3D SASHA agree well with IR-SE. In vivo experiments showed that the pre-contrast and post-contrast T1 maps acquired by 3D SACORA are comparable to the corresponding 3D SASHA maps, despite the shorter acquisition time (15s vs. 188s, for a heart rate of 60 bpm). Mean septal pre-contrast T1 was 1453 ± 44 ms with 3D SACORA and 1460 ± 60 ms with 3D SASHA. Mean septal post-contrast T1 was 824 ± 66 ms and 824 ± 60 ms. 3D SACORA acquires 3D T1 maps in 15 heart beats (heart rate, 60 bpm) at 3T. In addition to its short acquisition time, the sequence achieves good T1 estimation precision and accuracy.
Intestinal Epithelial Cell-Derived Extracellular Vesicles Modulate Hepatic Injury via the Gut-Liver Axis During Acute Alcohol Injury.
(Frontiers Media, 2020-12) Lamas-Paz, Arantza; Moran, Laura; Peng, Jin; Salinas, Beatriz; Lopez-Alcantara, Nuria; Sydor, Svenja; Vilchez-Vargas, Ramiro; Asensio, Iris; Hao, Fengjie; Zheng, Kang; Martin-Adrados, Beatriz; Moreno, Laura; Cogolludo, Angel; Gomez Del Moral, Manuel; Bechmann, Lars; Martinez-Naves, Eduardo; Vaquero, Javier; Bañares, Rafael; Nevzorova, Yulia A; Cubero, Francisco Javier; Consorcio NanoLiver-CM; Deutsche Forschungsgemeinschaft (Alemania); Chinese Scholarship Council; Natural Science Foundation of Jiangsu Province (China); Ministerio de Asuntos Económicos y Transformación Digital (España)
Binge drinking, i.e., heavy episodic drinking in a short time, has recently become an alarming societal problem with negative health impact. However, the harmful effects of acute alcohol injury in the gut-liver axis remain elusive. Hence, we focused on the physiological and pathological changes and the underlying mechanisms of experimental binge drinking in the context of the gut-liver axis. Eight-week-old mice with a C57BL/6 background received a single dose (p.o.) of ethanol (EtOH) [6 g/kg b.w.] as a preclinical model of acute alcohol injury. Controls received a single dose of PBS. Mice were sacrificed 8 h later. In parallel, HepaRGs and Caco-2 cells, human cell lines of differentiated hepatocytes and intestinal epithelial cells intestinal epithelial cells (IECs), respectively, were challenged in the presence or absence of EtOH [0-100 mM]. Extracellular vesicles (EVs) isolated by ultracentrifugation from culture media of IECs were added to hepatocyte cell cultures. Increased intestinal permeability, loss of zonula occludens-1 (ZO-1) and MUCIN-2 expression, and alterations in microbiota-increased Lactobacillus and decreased Lachnospiraceae species-were found in the large intestine of mice exposed to EtOH. Increased TUNEL-positive cells, infiltration of CD11b-positive immune cells, pro-inflammatory cytokines (e.g., tlr4, tnf, il1β), and markers of lipid accumulation (Oil Red O, srbep1) were evident in livers of mice exposed to EtOH, particularly in females. In vitro experiments indicated that EVs released by IECs in response to ethanol exerted a deleterious effect on hepatocyte viability and lipid accumulation. Overall, our data identified a novel mechanism responsible for driving hepatic injury in the gut-liver axis, opening novel avenues for therapy.
Harmine and Piperlongumine Revert TRIB2-Mediated Drug Resistance.
(Multidisciplinary Digital Publishing Institute (MDPI), 2020-12-09) Machado, Susana; Silva, Andreia; De Sousa-Coelho, Ana Luísa; Duarte, Isabel; Grenho, Ines; Santos, Bruno; Mayoral-Varo, Victor; Megias Vazquez, Diego; Sanchez-Cabo, Fatima; Dopazo, Ana; Ferreira, Bibiana I; Link, Wolfgang; Fundação para a Ciência e Tecnologia (Portugal); Ministerio de Ciencia, Innovación y Universidades (España); Unión Europea. Comisión Europea. H2020
Therapy resistance is responsible for most relapses in patients with cancer and is the major challenge to improving the clinical outcome. The pseudokinase Tribbles homologue 2 (TRIB2) has been characterized as an important driver of resistance to several anti-cancer drugs, including the dual ATP-competitive PI3K and mTOR inhibitor dactolisib (BEZ235). TRIB2 promotes AKT activity, leading to the inactivation of FOXO transcription factors, which are known to mediate the cell response to antitumor drugs. To characterize the downstream events of TRIB2 activity, we analyzed the gene expression profiles of isogenic cell lines with different TRIB2 statuses by RNA sequencing. Using a connectivity map-based computational approach, we identified drug-induced gene-expression profiles that invert the TRIB2-associated expression profile. In particular, the natural alkaloids harmine and piperlongumine not only produced inverse gene expression profiles but also synergistically increased BEZ235-induced cell toxicity. Importantly, both agents promote FOXO nuclear translocation without interfering with the nuclear export machinery and induce the transcription of FOXO target genes. Our results highlight the great potential of this approach for drug repurposing and suggest that harmine and piperlongumine or similar compounds might be useful in the clinic to overcome TRIB2-mediated therapy resistance in cancer patients.

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