Histone deacetylase 6 at crossroads of infection and innnate immunity

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de lectura: 25-05-2018HDAC6 is a tubulin deacetylase involved in many cellular functions related to cytoskeleton dynamics including cell migration and autophagy. In addition, HDAC6 affects antigen-dependent CD4+ T cell activation. In this study, we show that HDAC6 contributes to the cytotoxic function of CD8+ T cells. Immunization studies revealed defective cytotoxic activity in vivo in the absence of HDAC6. Adoptive transfer of wild-type or Hdac6-/- CD8+ T cells to Rag1-/- mice demonstrated specific impairment in CD8+ T cell responses against vaccinia infection. Mechanistically, HDAC6-deficient cytotoxic T lymphocytes (CTLs) showed defective in vitro cytolytic activity related to altered dynamics of lytic granules, inhibited kinesin 1 – dynactin mediated terminal transport of lytic granules to the immune synapse and deficient exocytosis, but not to target cell recognition, T cell receptor (TCR) activation or interferon (IFNγ) production. Our results establish HDAC6 as an effector of the immune cytotoxic response that acts by affecting the dynamics, transport and secretion of lytic granules by CTLsThis work has been performed at the Prof. Francisco Sánchez-Madrid’s laboratory in the Hospital Universitario de la Princesa (HUP) and Centro Nacional de Investigaciones Cardiovasculares (CNIC) in Madrid. This study was funded by grants SAF2014-55579-R from the Spanish Ministry of Economy and Competitiveness, INDISNET-S2011/ BMD-2332 from the Comunidad de Madrid, CIBERCARDIOVASCULAR and grant PIE13/00041 from the Instituto de Salud Carlos III (Fondo de Investigación Sanitaria del Instituto de Salud Carlos III with co-funding from the Fondo Europeo de Desarrollo Regional; FEDER), and ERC-2011-AdG 294340- GENTRIS and COST-Action BM1202 from the European Comission to Francisco Sánchez- Madrid. The Centro Nacional de Investigaciones Cardiovasculares (CNIC) is supported by the Spanish Ministry of Economy and Competitiveness (MINECO) and the Pro-CNIC Foundation and is a Severo Ochoa Center of Excellence (MINECO award SEV-2015-0505). Olga Moreno was supported by a PhD fellowship from the Spanish Ministry of Education (FPU Program, FPU12/00733). I thank Dr. M. Gómez for assitance with English editing and for critical reading of this PhD thesis

Landscape variables influencing forest fires in central Spain

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ISGylation controls exosome secretion by promoting lysosomal degradation of MVB proteins

Exosomes are vesicles secreted to the extracellular environment through fusion with the plasma membrane of specific endosomes called multivesicular bodies (MVB) and mediate cell-to-cell communication in many biological processes. Posttranslational modifications are involved in the sorting of specific proteins into exosomes. Here we identify ISGylation as a ubiquitin-like modification that controls exosome release. ISGylation induction decreases MVB numbers and impairs exosome secretion. Using ISG15-knockout mice and mice expressing the enzymatically inactive form of the de-ISGylase USP18, we demonstrate in vitro and in vivo that ISG15 conjugation regulates exosome secretion. ISG15 conjugation triggers MVB co-localization with lysosomes and promotes the aggregation and degradation of MVB proteins. Accordingly, inhibition of lysosomal function or autophagy restores exosome secretion. Specifically, ISGylation of the MVB protein TSG101 induces its aggregation and degradation, being sufficient to impair exosome secretion. These results identify ISGylation as a novel ubiquitin-like modifier in the control of exosome production.We thank Dr K. Knobeloch, Dr A. Garcia-Sastre and Dr M.A. Alonso for providing reagents, and Dr S. Bartlett for assistance with English editing. C.E. is thankful to electron microscopy facility (campus Casanova), CCiT-University of Barcelona. This study was supported by grants SAF2014-55579-R from the Spanish Ministry of Economy and Competitiveness, INDISNET-S2011/BMD-2332 from the Comunidad de Madrid, Cardiovascular Network RD12-0042-0056 and PIE13/00041 from Instituto de Salud Carlos III (Fondo de Investigacion Sanitaria del Instituto de Salud Carlos III and co-funding by Fondo Europeo de Desarrollo Regional FEDER), ERC-2011-AdG 294340-GENTRIS and COST-Action BM1202 to F.S.-M.; grant SAF2014-54623-R, FIS grant PI11/00127 (Fondo de Investigacion Sanitaria del Instituto de Salud Carlos III and Ministry of Health of Spain, State secretary of R+D and FEDER/FSE) and Bayer Group Grants4Grants (ID 2013-08-0982) to S.G.; and grant BFU2015-66785-P from the Spanish Ministry of Economy and Competitiveness to C.E.; Centro Nacional de Investigaciones Cardiovasculares (CNIC) is supported by the Spanish Ministry of Economy and Competitiveness (MINECO) and the Pro-CNIC Foundation, and is a Severo Ochoa Center of Excellence (MINECO award SEV-2015-0505). C.V.-B. was supported by FPU programme (Spanish Ministry of Education). M. M. is supported by MS14/00219 from Instituto de Salud Carlos III.S

Collective Intelligence to Find Solutions to the Challenges Posed by the Sustainable Development Goals

The implementation of the United Nations (UN) Sustainable Development Goals (SDGs) presents a vast and intricate array of challenges, including the establishment of governance systems that engage all societal actors, particularly nongovernmental entities and youth, in proposing solutions and decision-making. This article investigates the potential of collective intelligence as a tool within citizen science to create solutions for SDG-related challenges and to establish or enhance necessary governance mechanisms. We detail a collective intelligence experiment conducted during the UN Climate Change Conference 2019 (COP25; Madrid, December 2–13), which aimed to generate a prioritised list of actions addressing SDG 6, Water and Sanitation and SDG 13, Climate Action. The experiment involved 1,253 students aged 15 to 17 who proposed, modified, and prioritised 14,517 ideas using an online platform created by Kampal, a spin-off of the University of Zaragoza. We discuss: a) participation protocols following citizen science methodologies; b) the platform description; c) results concerning the participation process, the tool’s effectiveness in collectively extracting the best solutions, and the quality of the generated proposals; and d) enhancements and new research directions for using citizen science and collective intelligence to tackle SDG-related challenges in a collaborative and participatory way

One year overview and follow-up in a post-COVID consultation of critically ill patients

The long-term clinical management and evolution of a cohort of critical COVID-19 survivors have not been described in detail. We report a prospective observational study of COVID-19 patients admitted to the ICU between March and August 2020. The follow-up in a post-COVID consultation comprised symptoms, pulmonary function tests, the 6-minute walking test (6MWT), and chest computed tomography (CT). Additionally, questionnaires to evaluate the prevalence of post-COVID-19 syndrome were administered at 1 year. A total of 181 patients were admitted to the ICU during the study period. They were middle-aged (median [IQR] of 61 [52;67]) and male (66.9%), with a median ICU stay of 9 (5–24.2) days. 20% died in the hospital, and 39 were not able to be included. A cohort of 105 patients initiated the follow-up. At 1 year, 32.2% persisted with respiratory alterations and needed to continue the follow-up. Ten percent still had moderate/severe lung diffusion (DLCO) involvement (<60%), and 53.7% had a fibrotic pattern on CT. Moreover, patients had a mean (SD) number of symptoms of 5.7 ± 4.6, and 61.3% met the criteria for post-COVID syndrome at 1 year. During the follow-up, 46 patients were discharged, and 16 were transferred to other consultations. Other conditions, such as emphysema (21.6%), COPD (8.2%), severe neurocognitive disorders (4.1%), and lung cancer (1%) were identified. A high use of health care resources is observed in the first year. In conclusion, one-third of critically ill COVID-19 patients need to continue follow-up beyond 1 year, due to abnormalities on DLCO, chest CT, or persistent symptoms.This study was supported in part by ISCIII (CIBERESUCICOVID, COV20/00110), co-funded by ERDF, “Una manera de hacer Europa,” donation program “Estar Preparados,” UNESPA, Madrid, Spain and Fundación Soria Melguizo (Madrid, Spain). DG-C had received financial support from Instituto de Salud Carlos III (Miguel Servet 2020: CP20/00041), co-funded by the European Social Fund (ESF)/“Investing in your future.” JB acknowledged receiving financial support from Instituto de Salud Carlos III (ISCIII; Miguel Servet 2019: CP19/00108), co-funded by the European Social Fund (ESF), “Investing in your future.”Peer ReviewedArticle signat per 29 autors/es: Jessica González (1,2,3,4), María Zuil (1,2,3,4), Iván D. Benítez (2,3,4), David de Gonzalo-Calvo (2,3,4), María Aguilar (1,2), Sally Santisteve (1,2,3,4), Rafaela Vaca (1,2), Olga Minguez (1,2), Faty Seck (1,2), Gerard Torres (1,2,3,4), Jordi de Batlle (2,3,4), Silvia Gómez (1,2,3,4), Silvia Barril (1,2,3,4), Anna Moncusí-Moix (2,3,4), Aida Monge (1,2,3,4), Clara Gort-Paniello (2,3,4), Ricard Ferrer (4,5), Adrián Ceccato (4), Laia Fernández (4,6), Ana Motos (4,6), Jordi Riera (4,5), Rosario Menéndez (4,7), Darío Garcia-Gasulla (8), Oscar Peñuelas (4,9), Gonzalo Labarca (10,11), Jesús Caballero (12), Carme Barberà (13), Antoni Torres (4,6) and Ferran Barbé (1,2,3,4) * on behalf of the CIBERESUCICOVID Project (COV20/00110, ISCIII) // (1) Department of Pulmonary, Hospital Universitari Arnau de Vilanova and Santa Maria, Lleida, Spain, (2) Translational Research in Respiratory Medicine Group, Lleida, Spain, (3) Lleida Biomedical Research Institute, Lleida, Spain, (4) Centro de Investigación Biomédica en Red (CIBER) of Respiratory Diseases, Institute of Health Carlos III, Madrid, Spain, (5) Intensive Care Department, Vall d’Hebron Hospital Universitari, Shock, Organ Dysfunction and Resuscitation (SODIR) Research Group, Vall d’Hebron Institut de Recerca, Barcelona, Spain, (6) Department of Pulmonary, Hospital Clinic, Universitat de Barcelona, Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain, (7) Department of Pulmonary, University and Polytechnic Hospital La Fe, Valencia, Spain, (8) Barcelona Supercomputing Center, Barcelona, Spain, (9) Hospital Universitario de Getafe, Madrid, Spain, (10) Faculty of Medicine, University of Concepción, Concepción, Chile, (11) Department of Clinical Biochemistry and Immunology, Faculty of Pharmacy, Concepción, Chile, (12) Intensive Care Department, Hospital Universitari Arnau de Vilanova de Lleida, Lleida, Spain, (13) Intensive Care Department, Hospital Universitari Santa Maria de Lleida, Lleida, SpainPostprint (published version

HDAC6 regulates the dynamics of lytic granules in cytotoxic T lymphocytes

Journal of Cell Science.HDAC6 is a tubulin deacetylase involved in many cellular functions related to cytoskeleton dynamics, including cell migration and autophagy. In addition, HDAC6 affects antigen-dependent CD4+ T cell activation. In this study,we show that HDAC6 contributes to the cytotoxic function of CD8+ T cells. Immunization studies revealed defective cytotoxic activity in vivo in the absence of HDAC6. Adoptive transfer of wild-type or Hdac6-/- CD8+ T cells to Rag1-/- mice demonstrated specific impairment inCD8+ T cell responses against vaccinia infection. Mechanistically, HDAC6-deficient cytotoxic T lymphocytes (CTLs) showed defective in vitro cytolytic activity related to altered dynamics of lytic granules, inhibited kinesin-1-dynactin-mediated terminal transport of lytic granules to the immune synapse and deficient exocytosis, but not to target cell recognition, T cell receptor (TCR) activation or interferon (IFN)γ production. Our results establish HDAC6 as an effectorof theimmune cytotoxic response that acts byaffecting the dynamics, transport and secretion of lytic granules by CTLs.This work was supported by the Ministerio de Economı́a y competitividad (MINECO) [grant number SAF2014-55579-R]; Comunidad Autónoma de Madrid (CAM) [grant number INDISNET01592006]; Instituto de Salud Carlos III y Fondo Europeo de Desarrollo Regional (FEDER) [grant numbers BIOMID-PIE13/041 and RD12/0042/0056]; European Research Council (ERC) [grant number ERC-2011-AdG 294340- GENTRIS

The PDZ-adaptor protein syntenin-1 regulates HIV-1 entry

Syntenin-1 is a cytosolic adaptor protein involved in several cellular processes requiring polarization. Human immunodeficiency virus type 1 (HIV-1) attachment to target CD4(+) T-cells induces polarization of the viral receptor and coreceptor, CD4/CXCR4, and cellular structures toward the virus contact area, and triggers local actin polymerization and phosphatidylinositol 4,5-bisphosphate (PIP(2)) production, which are needed for successful HIV infection. We show that syntenin-1 is recruited to the plasma membrane during HIV-1 attachment and associates with CD4, the main HIV-1 receptor. Syntenin-1 overexpression inhibits HIV-1 production and HIV-mediated cell fusion, while syntenin depletion specifically increases HIV-1 entry. Down-regulation of syntenin-1 expression reduces F-actin polymerization in response to HIV-1. Moreover, HIV-induced PIP(2) accumulation is increased in syntenin-1–depleted cells. Once the virus has entered the target cell, syntenin-1 polarization toward the viral nucleocapsid is lost, suggesting a spatiotemporal regulatory role of syntenin-1 in actin remodeling, PIP(2) production, and the dynamics of HIV-1 entry

Extracellular vesicles from Listeria monocytogenes-infected dendritic cells alert the innate immune response.

Communication through cell-cell contacts and extracellular vesicles (EVs) enables immune cells to coordinate their responses against diverse types of pathogens. The function exerted by EVs in this context depends on the proteins and nucleic acids loaded into EVs, which elicit specific responses involved in the resolution of infection. Several mechanisms control protein and nucleic acid loading into EVs; in this regard, acetylation has been described as a mechanism of cellular retention during protein sorting to exosomes. HDAC6 is a deacetylase involved in the control of cytoskeleton trafficking, organelle polarity and cell migration, defense against Listeria monocytogenes (Lm) infection and other immune related functions. Here, we show that the protein content of dendritic cells (DCs) and their secreted EVs (DEVs) vary during Lm infection, is enriched in proteins related to antiviral functions compared to non-infected cells and depends on HDAC6 expression. Analyses of the post-translational modifications revealed an alteration of the acetylation and ubiquitination profiles upon Lm infection both in DC lysates and DEVs. Functionally, EVs derived from infected DCs upregulate anti-pathogenic genes (e.g. inflammatory cytokines) in recipient immature DCs, which translated into protection from subsequent infection with vaccinia virus. Interestingly, absence of Listeriolysin O in Lm prevents DEVs from inducing this anti-viral state. In summary, these data underscore a new mechanism of communication between bacteria-infected DC during infection as they alert neighboring, uninfected DCs to promote antiviral responses.This study was supported by grant PDI-2020-120412RB-I00, PDC2021-121797-I00, BIO2015-67580-P and PGC2018-097019-BI00 from the Spanish Ministry of Economy and Competitiveness (MINECO), grant S2017/BMD-3671-INFLAMUNE-CM from the Comunidad de Madrid, a grant from the Ramón Areces Foundation “Ciencias de la Vida y la Salud” (XIX Concurso-2018), “la Caixa” Banking Foundation (grants HR17-00016 and HR17-00247), BIOIMID (PIE13/041) and PRB3 (IPT17/0019 - ISCIII-SGEFI/ ERDF, ProteoRed) from Instituto de Salud Carlos III, CIBER Cardiovascular (CB16/11/00272), and Fondo de Investigación Sanitaria del Instituto de Salud Carlos III and co-funding by Fondo Europeo de Desarrollo Regional FEDER). IF-D is supported by a Fellowship from the Spanish Ministry of Science, Innovation, and Universities (FPU15/02539). DC-F is supported by a Fellowship from “la Caixa” Foundation (LCF/BQ/DR19/11740010). The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the Ministerio de Ciencia e Innovación (MCIN) and the Pro CNIC Foundation, and is a Severo Ochoa Center of Excellence (CEX2020- 001041-S). Funding agencies did not intervene in the design of the studies, with no copyright over the study.S

Interlaboratory study on Sb2S3 interplay between structure, dielectric function, and morphous-to-crystalline phase change for photonics

Antimony sulfide, Sb2S3, is interesting as the phase-change material for applications requiring high transmission from the visible to telecom wavelengths, with its band gap tunable from 2.2 to 1.6 eV, depending on the amorphous and crystalline phase. Here we present results from an interlaboratory study on the interplay between the structural change and resulting optical contrast during the amorphous-to-crystalline transformation triggered both thermally and optically. By statistical analysis of Raman and ellipsometric spectroscopic data, we have identified two regimes of crystallization, namely 250_C % T < 300_C, resulting in Type-I spherulitic crystallization yielding an optical contrast Dn _ 0.4, and 300 % T < 350 _ C, yielding Type-II crystallization bended spherulitic structure with different dielectric function and optical contrast Dn _ 0.2 below 1.5 eV. Based on our findings, applications of on-chip reconfigurable nanophotonic phase modulators and of a reconfigurable high-refractive-index core/phase-change shell nanoantenna are designed and proposed.The authors acknowledge the support from the European Union’s Horizon 2020 research and innovation program (No 899598 - PHEMTRONICS)

DataSheet_1_Extracellular vesicles from Listeria monocytogenes-infected dendritic cells alert the innate immune response.pdf [Dataset]

Supplementary Figure 1. Isolated EVs present typical size and topology. Supplementary Figure 2. Protein profiling from total cell lysates and their derived EVs from WT and KO-HDAC6 BMDCs. Supplementary Figure 3. Enrichment in acetylated and ubiquitinated DC proteins upon Lm infection. Supplementary Figure 4. Ubiquitination in K-48 and K-63 state in T lymphoblast total cell lysates and their derived EVs. Supplementary Figure 5. Pore filtration methods restrain Lm and do not induce strong antipathogenic responses. Supplementary Figure 6. IFN-β is detected following Lm infection. Table S1. List of antibodies used for Western-blot and Flow Cytometry and the used dilution. Table S2. List of primers, with their corresponding sequence, used for qPCR. Table S3: Protein quantification in total cell lysates Table S4: IPA analysis of total cell lysates: canonical pathways and diseases and functions category Table S5: Protein quantification in EVs Table S6: IPA analysis of EVs: diseases and functions category Table S7: Ubiquitinated and acetylated peptides in total cell lysates and EVs Table S8: Enrichment analysis of ubiquitinated and acetylated proteinsPeer reviewe