Descriptor: "MESH: Mice, Knockout" - Searchworks@Jio Institute Digital Library Search Results

Your search keyword '"MESH: Mice, Knockout"' showing total 908 results

Start Over Descriptor "MESH: Mice, Knockout"

908 results on '"MESH: Mice, Knockout"'

1. Evidence of the involvement of dystrophin Dp71 in corneal angiogenesis

Author: Ortiz, Gabriella, Vacca, Ophélie, Bénard, Romain, Dupas, Bénédicte, Sennlaub, Florian, Guillonneau, Xavier, JA, Sahel, Tadayoni, Ramin, Rendon, Alvaro, Giocanti-Aurégan, Audrey, Brighton and Sussex University Hospitals - NHS Trust (BSUH), Institut des Neurosciences Paris-Saclay (NeuroPSI), Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut de la Vision, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Hôpital Lariboisière-Fernand-Widal [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Diderot - Paris 7 (UPD7), Université Sorbonne Paris Cité (USPC), Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts (CHNO), Fondation Ophtalmologique Adolphe de Rothschild [Paris], Hôpital Avicenne [AP-HP], and PERIGNON, Alain
Subjects: genetic structures, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, [SDV.NEU.PC] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, MESH: Mice, Knockout, Cornea, Dystrophin, MESH: Dystrophin, Animals, Corneal Neovascularization, MESH: Animals, Aorta, Mice, Knockout, MESH: Corneal Injuries, MESH: Corneal Neovascularization, [SDV.NEU.PC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, MESH: Cornea, [SDV.NEU.NB] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, MESH: Aorta, eye diseases, Disease Models, Animal, sense organs, MESH: Disease Models, Animal, [SDV.NEU.SC] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, Research Article, Corneal Injuries
Abstract: International audience; Purpose: The aim of this study was to define the role of dystrophin Dp71 in corneal angiogenesis.Methods: Inflammation-induced corneal neovascularization experiments were performed in Dp71-null mice and C57BL/6J wild-type mice.Results: The corneal neovascular area covered by neovascularization was larger in the injured corneas of the Dp71-null mice compared to the corneas of the wild-type mice: 40.72% versus 26.33%, respectively (p
Published: 2022

2. Rare missense variant inMSH4associated with primary gonadal failure in both 46, XX and 46, XY individuals

Author: Mehdi Totonchi, Mehri Mashayekhi, Ken McElreavey, Navid Almadani, Anu Bashambou, Arvand Akbari, Kimiya Padidar, Najmeh Salehi, Mohammad Ali Sadighi Gilani, Royan Institute for Reproductive Biomedicine [Tehran, Iran], Academic Center for Education, Culture and Research (ACECR), University of Tehran, Génétique du Développement humain - Human developmental genetics, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), This study was financially supported by Royan Institute, Tehran, Iran, and Institut Pasteur, Paris, France, We are sincerely thankful to the personnel of Royan Department of Genetics and the Human Developmental Genetics Unit of Institut Pasteur, Paris, France, and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, Proband, 46, XX Disorders of Sex Development, [SDV]Life Sciences [q-bio], Cell Cycle Proteins, Iran, MESH: Mice, Knockout, Mice, non-obstructive azoospermia, Missense mutation, MESH: Animals, Exome sequencing, Mice, Knockout, Sanger sequencing, Genetics, 0303 health sciences, MESH: Paris, 030305 genetics & heredity, Rehabilitation, Obstetrics and Gynecology, oligozoospermia, DNA-Binding Proteins, symbols, Female, primary gonadal failure, France, Paris, dbSNP, familial exome sequencing, Biology, 03 medical and health sciences, symbols.namesake, MESH: Cell Cycle Proteins, Animals, Humans, MESH: Mice, Gene, Retrospective Studies, 030304 developmental biology, Disorder of Sex Development, 46,XY, MESH: Humans, MESH: Retrospective Studies, MSH4, MESH: Male, primary ovarian insufficiency, MESH: France, MSH5, Reproductive Medicine, MESH: Iran, MESH: Female, MESH: DNA-Binding Proteins
Abstract: STUDY QUESTIONCan whole-exome sequencing (WES) reveal a shared pathogenic variant responsible for primary gonadal failure in both male and female patients from a consanguineous family?SUMMARY ANSWERPatients with primary ovarian insufficiency (POI) and non-obstructive azoospermia (NOA) were homozygous for the rare missense variant p. S754L located in the highly conserved MSH4 MutS signature motif of the ATPase domain. An oligozoospermic patient was heterozygous for the variant.WHAT IS KNOWN ALREADYMSH4 is a meiosis-specific protein expressed at a certain level in the testes and ovaries. Along with its heterodimer partner MSH5, it is responsible for double-strand Holliday junction recognition and stabilization, to ensure accurate chromosome segregation during meiosis. Knockout male and female mice for Msh4 and Msh5 are reportedly infertile due to meiotic arrest. In humans, MSH4 is associated with male and female gonadal failure, with distinct variations in the MutS domain V.STUDY DESIGN, SIZE, DURATIONThis was a retrospective genetics study of a consanguineous family with multiple cases of gonadal failure in both genders. The subject family was recruited in Iran, in 2018.PARTICIPANTS/MATERIALS, SETTING, METHODSThe proband who is affected by POI, an NOA brother, a fertile sister and their parents were subjected to WES. The discovered variant was validated in these individuals, and the rest of the family was also genotyped by Sanger sequencing. The variant was not detected in 800 healthy Iranian individuals from the Iranome database nor in 30 sporadic NOA and 30 sporadic POI patients. Suggested effect in aberrant splicing was studied by RT-PCR. Moreover, protein homology modeling was used to further investigate the amino acid substitution in silico.MAIN RESULTS AND THE ROLE OF CHANCEThe discovered variant is very rare and has never been reported in the homozygous state. It occurs in the ATPase domain at Serine 754, the first residue within the highly conserved MutS signature motif, substituting it with a Leucine. All variant effect prediction tools indicated this variant as deleterious. Since the substitution occurs immediately before the Walker B motif at position 755, further investigations based on protein homology were conducted. Considering the modeling results, the nature of the substituted amino acid residue and the distances between p. S754L variation and the residues of the Walker B motif suggested the possibility of conformational changes affecting the ATPase activity of the protein.LARGE SCALE DATAWe have submitted dbSNP entry rs377712900 to ClinVar under SCV001169709, SCV001169708 and SCV001142647 for oligozoospermia, NOA and POI, respectively.LIMITATIONS, REASONS FOR CAUTIONStudies in model organisms can shed more light on the role of this variant as our results were obtained by variant effect prediction tools and protein homology modeling.WIDER IMPLICATIONS OF THE FINDINGSIdentification of variants in meiotic genes should improve genetic counseling for both male and female infertility. Also, as two of our NOA patients underwent testicular sperm extraction (TESE) with no success, ruling out the existence of pathogenic variants in meiotic genes in such patients prior to TESE could prove useful.STUDY FUNDING/COMPETING INTEREST(S)This study was financially supported by Royan Institute in Tehran, Iran, and Institut Pasteur in Paris, France. The authors declare no competing interests.TRIAL REGISTRATION NUMBERN/A
Published: 2021

3. Functional compensation precedes recovery of tissue mass following acute liver injury

Author: Ira Fleming, Carolyn Winston, Benjamin Kruft, Alex K. Shalek, Wolfram Goessling, Satdarshan P.S. Monga, Florian Mueller, Sungjin Ko, Udayan Apte, Chad Walesky, Kellie E. Kolb, Jake Henderson, Brigham and Women’s Hospital [Boston, MA], Harvard Medical School [Boston] (HMS), Institute of Medical Engineering & Science [Cambridge, MA] (IMES), Massachusetts Institute of Technology (MIT), Broad Institute of MIT and Harvard (BROAD INSTITUTE), Harvard Medical School [Boston] (HMS)-Massachusetts Institute of Technology (MIT)-Massachusetts General Hospital [Boston], Ragon Institute of MGH, MIT and Harvard, Koch Institute for Integrative Cancer Research at MIT [Cambridge, MA], University of Pittsburgh Medical Center [Pittsburgh, PA, États-Unis] (UPMC), Imagerie et Modélisation - Imaging and Modeling, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre de Bioinformatique, Biostatistique et Biologie Intégrative (C3BI), Department of Pharmacology, Toxicology, and Therapeutics [Kansas City, KS, USA], University of Kansas Medical Center [Kansas City, KS, USA], Harvard-MIT Division of Health Sciences and Technology [Cambridge], Dana-Farber Cancer Institute [Boston], Harvard Stem Cell Institute [Cambridge, USA] (HSCI), Harvard University, Massachusetts General Hospital [Boston, MA, USA], and C..W. is supported by F32DK111151, the Cholangiocarcinoma Foundation Research Fellowship, and the American Liver Foundation Charles Trey, MD Memorial Post-doctoral Research Fellowship. W.G. is supported by R01DK090311, R01DK105198, R24OD017870, and the Claudia Adams Barr Program for Excellence in Cancer Research. W.G. is a Pew Scholar in Biomedical Sciences. A.K.S. was supported, in part, by the Searle Scholars Program, the Beckman Young Investigator Program, a Sloan Fellowship in Chemistry, the NIH (1DP2GM1194192 & RM1HG0061931), and the MIT Stem Cell Initiative through Fondation MIT. U.A. is supported by R01DK098414. S.P.M is supported by NIH/NIDDK P30DK120531, Pittsburgh Liver Research Centre Grant, and NIH/NIDDK (DK62277, DK100287, and CA204586)
Subjects: 0301 basic medicine, MESH: beta Catenin, General Physics and Astronomy, MESH: Cell Cycle, MESH: Mice, Knockout, Biochemistry, Transcriptome, MESH: Hepatocytes, Mice, 0302 clinical medicine, Protein biosynthesis, MESH: Animals, lcsh:Science, Wnt Signaling Pathway, beta Catenin, Mice, Knockout, Multidisciplinary, Fluorescence in situ hybridization, Cell Cycle, MESH: Wnt Signaling Pathway, Wnt signaling pathway, Cell cycle, MESH: Hepatectomy, Liver regeneration, Cell biology, MESH: Wnt Proteins, Liver, 030211 gastroenterology & hepatology, MESH: Liver Regeneration, Reprogramming, Biotechnology, Science, MESH: Acetaminophen, Biology, General Biochemistry, Genetics and Molecular Biology, Article, 03 medical and health sciences, MESH: Gene Expression Profiling, MESH: Mice, Inbred C57BL, MESH: Cell Proliferation, Detoxification, Genetics, Animals, Hepatectomy, Transcriptomics, MESH: Mice, Molecular Biology, Acetaminophen, Cell Proliferation, Mechanism (biology), Gene Expression Profiling, Macrophages, MESH: Macrophages, General Chemistry, Regenerative process, Liver Regeneration, Gene expression profiling, Mice, Inbred C57BL, Wnt Proteins, Disease Models, Animal, 030104 developmental biology, Hepatocytes, lcsh:Q, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, Liver function, MESH: Disease Models, Animal, MESH: Liver
Abstract: The liver plays a central role in metabolism, protein synthesis and detoxification. It possesses unique regenerative capacity upon injury. While many factors regulating cellular proliferation during liver repair have been identified, the mechanisms by which the injured liver maintains vital functions prior to tissue recovery are unknown. Here, we identify a new phase of functional compensation following acute liver injury that occurs prior to cellular proliferation. By coupling single-cell RNA-seq with in situ transcriptional analyses in two independent murine liver injury models, we discover adaptive reprogramming to ensure expression of both injury response and core liver function genes dependent on macrophage-derived WNT/β-catenin signaling. Interestingly, transcriptional compensation is most prominent in non-proliferating cells, clearly delineating two temporally distinct phases of liver recovery. Overall, our work describes a mechanism by which the liver maintains essential physiological functions prior to cellular reconstitution and characterizes macrophage-derived WNT signals required for this compensation., The liver possesses the ability to regenerate following sudden injury. Here, the authors use single-cell RNA-sequencing and in situ transcriptional analyses to identify a new phase of liver regeneration in mice aimed at maintaining essential functions throughout the regenerative process.
Published: 2020

4. Targeting androgen signaling in ILC2s protects from IL-33–driven lung inflammation, independently of KLRG1

Author: Blanquart, Eve, Mandonnet, Audrey, Mars, Marion, Cenac, Claire, Anesi, Nina, Mercier, Pascale, Audouard, Christophe, Roga, Stephane, Serrano de Almeida, Gilberto, Bevan, Charlotte, Girard, Jean-Philippe, Pelletier, Lucette, Laffont, Sophie, Guéry, Jean-Charles, Institut Toulousain des Maladies Infectieuses et Inflammatoires (Infinity), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de pharmacologie et de biologie structurale (IPBS), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Centre de biologie du développement (CBD), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Imperial College London, laffont-pradines, sophie, Toulouse Institute for Infectious and Inflammatory Diseases, University of Toulouse, CNRS, INSERM, UPS, Toulouse, France. (Infinity), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, and Université Fédérale Toulouse Midi-Pyrénées
Subjects: Male, KLRG1, MESH: Signal Transduction, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH: Lung / pathology, androgen signaling, MESH: Mice, Knockout, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, ILC2, MESH: Testosterone / pharmacology, MESH: Pneumonia / immunology, MESH: Mice, Inbred C57BL, Animals, Lectins, C-Type, Testosterone, Sex differences in asthma, MESH: Animals, Lymphocytes, MESH: Pneumonia / pathology, Receptors, Immunologic, [SDV.IMM.ALL]Life Sciences [q-bio]/Immunology/Allergology, Lung, [SDV.IMM.II] Life Sciences [q-bio]/Immunology/Innate immunity, ComputingMilieux_MISCELLANEOUS, Mice, Knockout, Sex Characteristics, lung inflammation, MESH: Androgens / pharmacology, Pneumonia, MESH: Lung / immunology, Interleukin-33, MESH: Receptors, Immunologic / immunology, MESH: Male, Mice, Inbred C57BL, MESH: Lymphocytes / immunology, MESH: Lectins, C-Type / immunology, MESH: Interleukin-33 / immunology, Androgens, [SDV.IMM]Life Sciences [q-bio]/Immunology, Female, MESH: Female, [SDV.IMM.ALL] Life Sciences [q-bio]/Immunology/Allergology, Signal Transduction, MESH: Sex Characteristics
Abstract: International audience; Background: Allergic asthma is more severe and frequent in women than in men. In male mice, androgens negatively control group 2 innate lymphoid cell (ILC2) development and function by yet unknown mechanisms.Objectives: We sought to investigate the impact of androgen on ILC2 homeostasis and IL-33-mediated inflammation in female lungs. We evaluated the role of androgen receptor (AR) signaling and the contribution of the putative inhibitory receptor killer cell lectin-like receptor G1 (KLRG1).Methods: Subcutaneous pellets mimicking physiological levels of androgen were used to treat female mice together with mice expressing a reporter enzyme under the control of androgen response elements and mixed bone marrow chimeras to assess the cell-intrinsic role of AR activation within ILC2s. We generated KLRG1-deficient mice.Results: We established that lung ILC2s express a functionally active AR that can be in vivo targeted with exogenous androgens to negatively control ILC2 homeostasis, proliferation, and function. Androgen signaling upregulated KLRG1 on ILC2s, which inhibited their proliferation on E-cadherin interaction. Despite evidence that KLRG1 impaired the competitive fitness of lung ILC2s during inflammation, KLRG1 deficiency neither alters in vivo ILC2 numbers and functions, nor did it lead to hyperactive ILC2s in either sexes.Conclusions: AR agonists can be used in vivo to inhibit ILC2 homeostatic numbers and ILC2-dependent lung inflammation through cell-intrinsic AR activation. Although androgen signals in ILC2s to upregulate KLRG1, we demonstrate that KLRG1 is dispensable for androgen-mediated inhibition of pulmonary ILC2s.
Published: 2022

5. The anti-IgE mAb omalizumab induces adverse reactions by engaging Fcγ receptors

Author: Bianca Balbino, Julien Stackowicz, Kari C. Nadeau, Pauline Herviou, Ophélie Godon, Sébastien Brûlé, Delphine Sterlin, Andrew J. Murphy, Vera Voronina, Bruno Iannascoli, Odile Richard-Le Goff, Gaël A. Millot, Macdonald Lynn, Laurent L. Reber, Harris Faith, Pierre Bruhns, Anticorps en thérapie et pathologie - Antibodies in Therapy and Pathology, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Sorbonne Université (SU), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Biophysique Moléculaire (Plate-forme), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Regeneron Pharmaceuticals [Tarrytown], Stanford University, Centre de Physiopathologie Toulouse Purpan (CPTP), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), European Commission (Marie Skłodowska-Curie Individual Fellowship H2020-MSCA-IF-2014 656086)European 235Research Council (ERC)–Seventh Framework Program (ERC-2013-CoG 616050)Institut Pasteur initiative for valorizing the applications of research (ValoExpress 2017)INSERM, ATIP-Avenir program'Investissement d’Avenir'(grant ANR-11-INBS-0006)Pasteur -Paris University (PPU) International Ph.DprogramFrench 'Fondation pour la Recherche Médicale FRMAP-HP, ANR-11-INBS-0006,FLI,France Life Imaging(2011), European Project: 616050,EC:FP7:ERC,ERC-2013-CoG,MYELOSHOCK(2014), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Service d'Immunologie [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: 0301 basic medicine, Allergy, MESH: Asthma / immunology, Omalizumab, MESH: Anaphylaxis / immunology, Immunoglobulin E, MESH: Mice, Knockout, Mice, MESH: Asthma / pathology, 0302 clinical medicine, Medicine, MESH: Animals, MESH: Drug Eruptions / pathology, Mice, Knockout, biology, Concise Communication, General Medicine, MESH: Drug Eruptions / genetics, 3. Good health, MESH: Anaphylaxis / pathology, MESH: Omalizumab / genetics, 030220 oncology & carcinogenesis, MESH: Drug Eruptions / immunology, [SDV.IMM]Life Sciences [q-bio]/Immunology, Drug Eruptions, Antibody, medicine.symptom, Anaphylaxis, medicine.drug, MESH: Mutation, MESH: Anaphylaxis / chemically induced, medicine.drug_class, Immunology, Immunoglobulins, Inflammation, Monoclonal antibody, MESH: Receptors, IgG / immunology, 03 medical and health sciences, Immune system, MESH: Omalizumab / adverse effects, Animals, MESH: Receptors, IgG / genetics, MESH: Mice, MESH: Omalizumab / pharmacology, business.industry, Receptors, IgG, MESH: Asthma / drug therapy, medicine.disease, Asthma, 030104 developmental biology, Mutation, Mast cells, MESH: Anaphylaxis / genetics, biology.protein, business
Abstract: International audience; Omalizumab is an anti-IgE monoclonal antibody (mAb) approved for the treatment of severe asthma and chronic spontaneous urticaria. Use of Omalizumab is associated with reported side effects, ranging from local skin inflammation at the injection site to systemic anaphylaxis. To date, the mechanisms through which Omalizumab induces adverse reactions are still unknown. Here, we demonstrated that immune complexes formed between Omalizumab and IgE can induce both skin inflammation and anaphylaxis through engagement of IgG receptors (FcγRs) in FcγR-humanized mice. We further developed an Fc-engineered mutant version of Omalizumab, and demonstrated that this mAb is equally potent as Omalizumab at blocking IgE-mediated allergic reactions, but does not induce FcγR-dependent adverse reactions. Overall, our data indicate that Omalizumab can induce skin inflammation and anaphylaxis by engaging FcγRs, and demonstrate that Fc-engineered versions of the mAb could be used to reduce such adverse reactions.
Published: 2020

6. House dust mites activate nociceptor–mast cell clusters to drive type 2 skin inflammation

Author: Mindy Tsai, Camille Petitfils, Nicolas Cenac, Stephen J. Galli, Riccardo Sibilano, Xinzhong Dong, Laurent L. Reber, Nicolas Gaudenzio, Lilian Basso, Philipp Starkl, Nadine Serhan, Chrystelle Bonnart, Thomas Marichal, James Meixiong, Institut National de la Santé et de la Recherche Médicale (INSERM), Stanford University School of Medicine [CA, USA], Institut de Recherche en Santé Digestive (IRSD ), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Johns Hopkins University, School of Medicine, Centre de Physiopathologie Toulouse Purpan (CPTP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Liège, Walloon Excellence in Life sciences and BIOtechnology [Liège] (WELBIO), Research Center for Molecular Medicine of the Austrian Academy of Sciences, Medizinische Universität Wien = Medical University of Vienna, the Stanford Neuroscience Microscopy Service (supported by NIH No. NS069375), (IFR30, Plateau Technique Cytometrie, Toulouse), (IFR30, Plateau Technique Imagerie Cellulaire, Toulouse), ‘Incentive Grant for Scientific Research’ of the F.R.S.-FNRS (No. F.4508.18), by the FRFS-WELBIO under grant No. CR-2017 s-04, by the Acteria Foundation and by an ERC Starting Grant (No. IM-ID 801823), the Austrian Science Fund (No. P31113-B30), ProdInra, Migration, Institut National de la Recherche Agronomique (INRA)-Université Toulouse III - Paul Sabatier (UT3), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT)
Subjects: Male, 0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, MESH: Nociceptors / metabolism, Dermatitis, MESH: Skin / cytology, Cell Communication, MESH: Mast Cells / metabolism, MESH: Tachykinins / genetics, MESH: Dermatitis, Atopic / pathology, MESH: Mice, Knockout, Receptors, G-Protein-Coupled, Mice, 0302 clinical medicine, TAC1, Receptors, 2.1 Biological and endogenous factors, Immunology and Allergy, Mast Cells, Aetiology, Acute inflammation, Skin, Mice, Knockout, MESH: Cell Communication / immunology, Chemistry, MESH: Dermatitis, Atopic / immunology, Pyroglyphidae, Pain Research, Degranulation, Nociceptors, Mast cell, Research Highlight, MESH: Mast Cells / immunology, 3. Good health, Nociception, medicine.anatomical_structure, Nociceptor, MESH: Tachykinins / metabolism, Female, Chronic Pain, medicine.symptom, Cell signalling, MESH: Skin / immunology, Knockout, Immunology, TRPV1, TRPV Cation Channels, Inflammation, MESH: Nociceptors / immunology, Article, Atopic, Dermatitis, Atopic, G-Protein-Coupled, 03 medical and health sciences, Tachykinins, medicine, Animals, Humans, MESH: Receptors, G-Protein-Coupled / metabolism, Animal, Inflammatory and immune system, Neurosciences, MESH: Allergens / immunology, MESH: TRPV Cation Channels / metabolism, MESH: Pyroglyphidae / immunology, Allergens, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, [SDV.AEN] Life Sciences [q-bio]/Food and Nutrition, Disease Models, Animal, 030104 developmental biology, nervous system, Disease Models, Itching, MESH: Disease Models, Animal, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, 030215 immunology
Abstract: International audience; Allergic skin diseases, such as atopic dermatitis, are clinically characterized by severe itching and type 2 immunity-associated hypersensitivity to widely distributed allergens, including those derived from house dust mites (HDMs). Here we found that HDMs with cysteine protease activity directly activated peptidergic nociceptors, which are neuropeptide-producing nociceptive sensory neurons that express the ion channel TRPV1 and Tac1, the gene encoding the precursor for the neuropeptide substance P. Intravital imaging and genetic approaches indicated that HDM-activated nociceptors drive the development of allergic skin inflammation by inducing the degranulation of mast cells contiguous to such nociceptors, through the release of substance P and the activation of the cationic molecule receptor MRGPRB2 on mast cells. These data indicate that, after exposure to HDM allergens, activation of TRPV1+Tac1+ nociceptor–MRGPRB2+ mast cell sensory clusters represents a key early event in the development of allergic skin reactions.
Published: 2019

7. SHP2 drives inflammation-triggered insulin resistance by reshaping tissue macrophage populations

Author: Ophélie Pereira, Jean Personnaz, Céline Saint-Laurent, Toshiyuki Araki, Maithé Tauber, Jean-Pierre Salles, Philippe Valet, Adélaïde Gélineau, Mickaël Canouil, Romain Paccoud, Armelle Yart, Jacques Weill, Jean-Philippe Pradere, Mylène Tajan, Cédric Dray, Enzo Piccolo, Sophie Le Gonidec, Inès Baba, Thomas Edouard, Emmanuel L. Gautier, Haoussa Askia, Philippe Froguel, Benjamin G. Neel, Nicolas Beton, Simon Deleruyelle, Johanna Auriau, Sophie Branka, Alizée Dortignac, Maxime Branchereau, Isabelle Castan, Christophe Heymes, Rémy Burcelin, Julie Charpentier, Institut des Maladies Métaboliques et Casdiovasculaires (UPS/Inserm U1297 - I2MC), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Geroscience and rejuvenation research center (RESTORE), Université de Toulouse (UT)-Université de Toulouse (UT)-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut Européen de Génomique du Diabète - European Genomic Institute for Diabetes - FR 3508 (EGID), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Centre de Physiopathologie Toulouse Purpan (CPTP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), ANR-17-CE14-0016,HOLISTic,Rôle de la protéine HMGB1 au cours du stress metabolique(2017), ANR-17-CE14-0009,CAPTOR,Contrôler la fonction des progéniteurs du tissu adipeux pour améliorer les désordres métaboliques de l'obésité(2017), and ANR-17-CE14-0023,TARGETKC,Homéostasie et rôles des cellules de Kupffer dans la stéatohépatite(2017)
Subjects: MESH: Inflammation, medicine.medical_specialty, endocrine system diseases, education, Inflammation, Type 2 diabetes, MESH: Mice, Knockout, Diabetes Mellitus, Experimental, Proinflammatory cytokine, Mice, 03 medical and health sciences, 0302 clinical medicine, Insulin resistance, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Internal medicine, MESH: Diabetes Mellitus, Experimental, medicine, Animals, Humans, Glucose homeostasis, MESH: Animals, Macrophage homeostasis, MESH: Mice, 030304 developmental biology, Mice, Knockout, 0303 health sciences, MESH: Humans, business.industry, Macrophages, Monocyte, nutritional and metabolic diseases, MESH: Macrophages, General Medicine, medicine.disease, 3. Good health, MESH: Insulin Resistance, medicine.anatomical_structure, Endocrinology, Adipose Tissue, Diabetes Mellitus, Type 2, Lipotoxicity, Insulin Resistance, medicine.symptom, business, MESH: Adipose Tissue, 030217 neurology & neurosurgery, MESH: Diabetes Mellitus, Type 2
Abstract: Insulin resistance is a key event in type 2 diabetes onset and a major comorbidity of obesity. It results from a combination of fat excess-triggered defects, including lipotoxicity and metaflammation, but the causal mechanisms remain difficult to identify. Here, we report that hyperactivation of the tyrosine phosphatase SHP2 found in Noonan syndrome (NS) led to an unsuspected insulin resistance profile uncoupled from altered lipid management (for example, obesity or ectopic lipid deposits) in both patients and mice. Functional exploration of an NS mouse model revealed this insulin resistance phenotype correlated with constitutive inflammation of tissues involved in the regulation of glucose metabolism. Bone marrow transplantation and macrophage depletion improved glucose homeostasis and decreased metaflammation in the mice, highlighting a key role of macrophages. In-depth analysis of bone marrow-derived macrophages in vitro and liver macrophages showed that hyperactive SHP2 promoted a proinflammatory phenotype, modified resident macrophage homeostasis, and triggered monocyte infiltration. Consistent with a role of SHP2 in promoting inflammation-driven insulin resistance, pharmaceutical SHP2 inhibition in obese diabetic mice improved insulin sensitivity even better than conventional antidiabetic molecules by specifically reducing metaflammation and alleviating macrophage activation. Together, these results reveal that SHP2 hyperactivation leads to inflammation-triggered metabolic impairments and highlight the therapeutical potential of SHP2 inhibition to ameliorate insulin resistance.
Published: 2021

8. Identification of Acer2 as a First Susceptibility Gene for Lithium-Induced Nephrogenic Diabetes Insipidus in Mice

Author: Cungui Mao, Birgitte Mønster Christensen, Ron Korstanje, Alain Doucet, Karolina M. Andralojc, Benjamin E. Low, Roger Sandhoff, Peter M.T. Deen, Michael V. Wiles, Lydie Cheval, Lena K. Ebert, Theun de Groot, Ruben Baumgarten, Radboud university [Nijmegen], Centre de Recherche des Cordeliers (CRC), Université Pierre et Marie Curie - Paris 6 (UPMC)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), and German Cancer Research Center - Deutsches Krebsforschungszentrum [Heidelberg] (DKFZ)
Subjects: [SDV]Life Sciences [q-bio], lnfectious Diseases and Global Health Radboud Institute for Molecular Life Sciences [Radboudumc 4], 030232 urology & nephrology, MESH: Mice, Knockout, 0302 clinical medicine, Inbred strain, MESH: Sodium, MESH: Animals, Acidosis, 0303 health sciences, MESH: Dinoprostone, MESH: Genetic Predisposition to Disease, MESH: Lithium Chloride, General Medicine, water-electrolyte balance, 3. Good health, Nephrology, Urine osmolality, acidosis, medicine.symptom, medicine.medical_specialty, chemistry.chemical_element, genetics and development, MESH: Acidosis, Calcium, MESH: Mice, Inbred Strains, MESH: Hypercalcemia, MESH: Hematocrit, 03 medical and health sciences, Internal medicine, medicine, MESH: Species Specificity, MESH: Diabetes Insipidus, Nephrogenic, Adverse effect, MESH: Kidney Tubules, Collecting, MESH: Mice, MESH: Nephrons, MESH: RNA, Messenger, 030304 developmental biology, Calcium metabolism, calcium, MESH: Alkaline Ceramidase, business.industry, medicine.disease, Nephrogenic diabetes insipidus, MESH: Acid-Base Equilibrium, Renal disorders Radboud Institute for Molecular Life Sciences [Radboudumc 11], Endocrinology, chemistry, MESH: Genome-Wide Association Study, Diabetes insipidus, business, MESH: Female
Abstract: Item does not contain fulltext BACKGROUND: Lithium, mainstay treatment for bipolar disorder, causes nephrogenic diabetes insipidus and hypercalcemia in about 20% and 10% of patients, respectively, and may lead to acidosis. These adverse effects develop in only a subset of patients treated with lithium, suggesting genetic factors play a role. METHODS: To identify susceptibility genes for lithium-induced adverse effects, we performed a genome-wide association study in mice, which develop such effects faster than humans. On day 8 and 10 after assigning female mice from 29 different inbred strains to normal chow or lithium diet (40 mmol/kg), we housed the animals for 48 hours in metabolic cages for urine collection. We also collected blood samples. RESULTS: In 17 strains, lithium treatment significantly elevated urine production, whereas the other 12 strains were not affected. Increased urine production strongly correlated with lower urine osmolality and elevated water intake. Lithium caused acidosis only in one mouse strain, whereas hypercalcemia was found in four strains. Lithium effects on blood pH or ionized calcium did not correlate with effects on urine production. Using genome-wide association analyses, we identified eight gene-containing loci, including a locus containing Acer2, which encodes a ceramidase and is specifically expressed in the collecting duct. Knockout of Acer2 led to increased susceptibility for lithium-induced diabetes insipidus development. CONCLUSIONS: We demonstrate that genome-wide association studies in mice can be used successfully to identify susceptibility genes for development of lithium-induced adverse effects. We identified Acer2 as a first susceptibility gene for lithium-induced diabetes insipidus in mice. 01 december 2019
Published: 2019

9. ANP-stimulated Na+ secretion in the collecting duct prevents Na+ retention in the renal adaptation to acid load

Author: Alain Doucet, Geneviève Escher, Luciana Morla, Gabrielle Planelles, Dignê Tembely, Lydie Cheval, Christine Walter, Bruno Vogt, Gilles Crambert, Naziha Bakouh, Centre de Recherche des Cordeliers (CRC), Université Paris Diderot - Paris 7 (UPD7)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Métabolisme et physiologie rénales (ERL 8228), Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Régulation des systèmes de transport dans les épithéliums (UMR_S467), and Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: metabolic acidosis, MESH: Signal Transduction, 0301 basic medicine, MESH: Hydrogen-Ion Concentration, MESH: Atrial Natriuretic Factor, Physiology, [SDV]Life Sciences [q-bio], Stimulation, MESH: Mice, Knockout, chemistry.chemical_compound, 0302 clinical medicine, Atrial natriuretic peptide, MESH: Sodium, MESH: Cyclic GMP, MESH: H(+)-K(+)-Exchanging ATPase, MESH: Animals, 610 Medicine & health, Aldosterone, Second messenger system, hormones, hormone substitutes, and hormone antagonists, medicine.medical_specialty, MESH: Rats, medicine.drug_class, H-K-ATPase, MESH: Acidosis, collecting duct, 03 medical and health sciences, MESH: Mice, Inbred C57BL, MESH: Xenopus laevis, Internal medicine, atrial natriuretic peptide, medicine, Intercalated Cell, MESH: Paracrine Communication, MESH: Kidney Tubules, Collecting, aldosterone, MESH: Aldosterone, Metabolic acidosis, medicine.disease, MESH: Adaptation, Physiological, cGMP, sodium secretion, MESH: Acid-Base Equilibrium, 030104 developmental biology, Endocrinology, chemistry, Mineralocorticoid, 570 Life sciences, biology, Cotransporter, MESH: Female, 030217 neurology & neurosurgery
Abstract: We have recently reported that type A intercalated cells of the collecting duct secrete Na+ by a mechanism coupling the basolateral type 1 Na+-K+-2Cl− cotransporter with apical type 2 H+-K+-ATPase (HKA2) functioning under its Na+/K+ exchange mode. The first aim of the present study was to evaluate whether this secretory pathway is a target of atrial natriuretic peptide (ANP). Despite hyperaldosteronemia, metabolic acidosis is not associated with Na+ retention. The second aim of the present study was to evaluate whether ANP-induced stimulation of Na+ secretion by type A intercalated cells might account for mineralocorticoid escape during metabolic acidosis. In Xenopus oocytes expressing HKA2, cGMP, the second messenger of ANP, increased the membrane expression, activity, and Na+-transporting rate of HKA2. Feeding mice with a NH4Cl-enriched diet increased urinary excretion of aldosterone and induced a transient Na+ retention that reversed within 3 days. At that time, expression of ANP mRNA in the collecting duct and urinary excretion of cGMP were increased. Reversion of Na+ retention was prevented by treatment with an inhibitor of ANP receptors and was absent in HKA2-null mice. In conclusion, paracrine stimulation of HKA2 by ANP is responsible for the escape of the Na+-retaining effect of aldosterone during metabolic acidosis.
Published: 2019

10. EZH2 cooperates with E2F1 to stimulate expression of genes involved in adrenocortical carcinoma aggressiveness

Author: Bruno Ragazzon, Jean-Christophe Pointud, Houda Tabbal, Pierre Val, Igor Tauveron, Coralie Drelon, M. Batisse-Lignier, Cyril Djari, Stéphanie Rodriguez, Anne-Marie Lefrançois-Martinez, Amandine Septier, Jérôme Bertherat, Guillaume Assié, Isabelle Sahut-Barnola, Mickael Mathieu, Antoine Martinez, Génétique, Reproduction et Développement (GReD), Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Institut National de la Santé et de la Recherche Médicale (INSERM), Génétique, Reproduction et Développement (GReD ), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), CHU Clermont-Ferrand, Institut Cochin (UMR_S567 / UMR 8104), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre National de la Recherche Scientifique (CNRS), Génétique, Reproduction et Développement - Clermont Auvergne (GReD ), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne (UCA)-Centre National de la Recherche Scientifique (CNRS), Génétique, Reproduction et Développement - Clermont Auvergne (GReD), Institut Cochin (IC UM3 (UMR 8104 / U1016)), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)
Subjects: Cancer Research, Adenosine, Indoles, Cell Cycle Proteins, MESH: Adrenal Cortex Neoplasms, MESH: Mice, Knockout, MESH: Proportional Hazards Models, MESH: Adrenocortical Carcinoma, 0302 clinical medicine, Gene expression, Adrenocortical Carcinoma, E2F1, MESH: Animals, Mice, Knockout, MESH: Chromatin Immunoprecipitation, MESH: Indoles, Regulation of gene expression, biology, EZH2, MESH: Gene Expression Regulation, Neoplastic, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, MESH: Ribonucleoside Diphosphate Reductase, 3. Good health, Gene Expression Regulation, Neoplastic, Securin, MESH: E2F1 Transcription Factor, Histone, Oncology, 030220 oncology & carcinogenesis, MESH: Computational Biology, Chromatin Immunoprecipitation, Ribonucleoside Diphosphate Reductase, [SDV.CAN]Life Sciences [q-bio]/Cancer, macromolecular substances, Article, Adrenal tumours, MESH: Multivariate Analysis, 03 medical and health sciences, MESH: Cell Cycle Proteins, Cancer epigenetics, MESH: Cell Proliferation, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Enhancer of Zeste Homolog 2 Protein, Animals, Humans, Enhancer of Zeste Homolog 2 Protein, Transcription factor, Cell Proliferation, Proportional Hazards Models, MESH: Humans, Cell growth, Computational Biology, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH: Adenosine, Adrenal Cortex Neoplasms, MESH: Securin, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Multivariate Analysis, biology.protein, Cancer research, Chromatin immunoprecipitation, E2F1 Transcription Factor
Abstract: International audience; BACKGROUND: EZH2 is overexpressed and associated with poor prognosis in adrenocortical carcinoma (ACC) and its inhibition reduces growth and aggressiveness of ACC cells in culture. Although EZH2 was identified as the methyltransferase that deposits the repressive H3K27me3 histone mark, it can cooperate with transcription factors to stimulate gene transcription. METHODS: We used bioinformatics approaches on gene expression data from three cohorts of patients and a mouse model of EZH2 ablation, to identify targets and mode of action of EZH2 in ACC. This was followed by ChIP and functional assays to evaluate contribution of identified targets to ACC pathogenesis. RESULTS: We show that EZH2 mostly works as a transcriptional inducer in ACC, through cooperation with the transcription factor E2F1 and identify three positive targets involved in cell cycle regulation and mitosis i.e., RRM2, PTTG1 and ASE1/PRC1. Overexpression of these genes is associated with poor prognosis, suggesting a potential role in acquisition of aggressive ACC features. Pharmacological and siRNA-mediated inhibition of RRM2 blocks cell proliferation, induces apoptosis and inhibits cell migration, suggesting that it may be an interesting target in ACC. CONCLUSIONS: Altogether, these data show an unexpected role of EZH2 and E2F1 in stimulating expression of genes associated with ACC aggressiveness. British Journal of Cancer https://doi.org/10.1038/s41416-019-0538-y BACKGROUND Adrenocortical carcinomas (ACC) are endocrine malignancies associated with dismal prognosis. At diagnosis, 80% of patients present metastases reducing the 5 years survival rate below 30% for most series.
Published: 2019

11. Muc5b is mainly expressed and sialylated in the nasal olfactory epithelium whereas Muc5ac is exclusively expressed and fucosylated in the nasal respiratory epithelium

Author: Jean-Luc Desseyn, Valérie Gouyer, Frédéric Gottrand, Céline Portal, Salah-Eddine Amini, Lille Inflammation Research International Center - U 995 (LIRIC), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Johns Hopkins University (JHU), S.-E. A. is the recipient of a 'Université de Lille' fellowship. C.P. was the recipient of a 'Université Lille 2' fellowship., We thank J. Bara for the gift of the 45M1 antibody, M.H. Gevaert (Service Commun-Morphologie Cellulaire, Université de Lille) for the slides, J. Devassine and Y. Lepage from the EOPS animal facility (Université de Lille) for mouse colony management and S. Plet and M. Magnien (LIRIC) for genotyping and dissection and the Bio-imaging Center (plate-forme de microscopie photonique, Université Lille) for Carl Zeiss Axio Scan. Z1 scanner., Lille Inflammation Research International Center (LIRIC), Université de Lille, Inserm, CHU Lille, and Lille Inflammation Research International Center - U 995 [LIRIC]
Subjects: 0301 basic medicine, Glycosylation, [SDV]Life Sciences [q-bio], Mucin 5AC, MESH: Mucin 5AC/analysis, MESH: Respiratory Mucosa/cytology, MESH: Mice, Knockout, Mice, MESH: Animals, MESH: Mucin-5B/metabolism, Receptor, Muc5b, Nose, Mice, Knockout, biology, Chemistry, MESH: Mucin-5B/deficiency, MESH: Nasal Mucosa/chemistry, respiratory system, MESH: Glycosylation, Immunohistochemistry, Mucin-5B, 3. Good health, Cell biology, Medical Laboratory Technology, medicine.anatomical_structure, Mouse nose, MESH: Mucin 5AC/genetics, Muc5ac, MESH: Respiratory Mucosa/metabolism, Histology, Respiratory Mucosa, 03 medical and health sciences, MESH: Mice, Inbred C57BL, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], medicine, Animals, Bowman’s gland, MESH: Mice, Molecular Biology, MESH: Nasal Mucosa/cytology, MESH: Nasal Mucosa/metabolism, Lamina propria, 030102 biochemistry & molecular biology, MESH: Mucin-5B/analysis, Mucin, Lectin, MESH: Immunohistochemistry, Cell Biology, Mucus, Mice, Inbred C57BL, Nasal Mucosa, 030104 developmental biology, biology.protein, Respiratory epithelium, MESH: Respiratory Mucosa/chemistry, MESH: Mucin 5AC/metabolism, Olfactory epithelium
Abstract: International audience; The nose is a complex organ that filters and warms breathing airflow. The nasal epithelium is the first barrier between the host and the external environment and is covered by a mucus gel that is poorly documented. Mucins are large, heavily O-glycosylated polymeric molecules secreted in the nose lumen by specialized cells, and they are responsible for the biochemical properties of the mucus gel. The mucus traps particles and clears them, and it also bathes microbiota, host molecules, and receptors that are all essential for odor perception in the olfactory epithelium. We used histology and immunohistochemistry to study the expression of the two main airway polymeric mucins, Muc5ac and Muc5b, in wild-type, green fluorescent protein-reporter Muc5b, and in genetically Muc5b-deficient mice. We report that Muc5ac is produced by goblet cells at the cell surface in the respiratory epithelium but is not expressed in the olfactory epithelium, whereas Muc5b is secreted by Bowman's glands situated in the lamina propria beneath the olfactory epithelium and also by goblet cells in the distal part of the respiratory epithelium. We also observed that Muc5b-deficient mice exhibited depletion of Bowman's glands. Using lectins, we found that terminally O-glycosylated chains of Muc5b were sialylated but not fucosylated, whereas Muc5ac was fucosylated but not sialylated. Specific localization and specific terminal glycosylation of the two mucins suggest different functions of the mucins.
Published: 2019

12. Orthobunyavirus spike architecture and recognition by neutralizing antibodies

Author: Jan Hellert, Martin Beer, Ahmed Haouz, Kerstin Wernike, Félix A. Rey, Sven Reiche, Pablo Guardado-Calvo, Emiliana Brocchi, Andrea Aebischer, Virologie Structurale - Structural Virology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institute of Diagnostic Virology (IVD), Friedrich-Loeffler-Institut (FLI), Cristallographie (Plateforme) - Crystallography (Platform), Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna 'Bruno Ubertini' (IZSLER), This research was performed as part of the Zoonoses Anticipation and Preparedness Initiative (ZAPI project, IMI Grant Agreement n°115760), with the assistance and financial support of IMI and the European Commission, and in kind contributions from EFPIA partners. F.A.R. also received funding by the Région Ile de France (Domaine d’intêret majeur Innovative technologies for life sciences, DIM 1HEALTH). Additional funding was provided by Institut Pasteur, the CNRS and the GIS IBiSA (Infrastructures en biologie santé et agronomie). J.H. received the Pasteur-Cantarini fellowship for 24 months, and was later supported by the DIM 1HEALTH grant., We thank Patrick England from the Molecular Biophysics facility and Fabrice Agou from the Chemogenomic and Biological Screening platform at Institut Pasteur and the staff of synchrotron beamlines PX1 and PX2 at SOLEIL (St. Aubin, France) and ID29 and ID30B at the ESRF (Grenoble, France) for help during data collection. We thank Xiaohong Shi and Richard M. Elliott from Glasgow, UK, for the BUNV Gc gene., and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, 0301 basic medicine, Orthobunyavirus, General Physics and Astronomy, MESH: Cricetinae, 02 engineering and technology, MESH: Amino Acid Sequence, Crystallography, X-Ray, medicine.disease_cause, Genome, MESH: Mice, Knockout, MESH: Antibodies, Monoclonal, MESH: Antibodies, Neutralizing, Mice, MESH: Protein Structure, Tertiary, Viral Envelope Proteins, MESH: Chlorocebus aethiops, Cricetinae, Chlorocebus aethiops, MESH: Animals, Neutralizing antibody, lcsh:Science, Mice, Knockout, Multidisciplinary, biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Antibodies, Monoclonal, Schmallenberg virus, Ruminants, 021001 nanoscience & nanotechnology, 3. Good health, MESH: Ruminants, Female, Antibody, 0210 nano-technology, Viral protein, Science, MESH: Glycoproteins, MESH: Orthobunyavirus, MESH: Vero Cells, Article, General Biochemistry, Genetics and Molecular Biology, Antigenic drift, 03 medical and health sciences, MESH: Viral Structures, MESH: Mice, Inbred C57BL, medicine, Animals, Amino Acid Sequence, Vero Cells, MESH: Mice, Viral Structures, Glycoproteins, General Chemistry, biology.organism_classification, MESH: Crystallography, X-Ray, Antibodies, Neutralizing, Virology, MESH: Male, Protein Structure, Tertiary, Mice, Inbred C57BL, 030104 developmental biology, MESH: Viral Envelope Proteins, biology.protein, Vero cell, lcsh:Q, MESH: Female
Abstract: Orthobunyaviruses (OBVs) form a distinct genus of arthropod-borne bunyaviruses that can cause severe disease upon zoonotic transmission to humans. Antigenic drift or genome segment re-assortment have in the past resulted in new pathogenic OBVs, making them potential candidates for causing emerging zoonoses in the future. Low-resolution electron cryo-tomography studies have shown that OBV particles feature prominent trimeric spikes, but their molecular organization remained unknown. Here we report X-ray crystallography studies of four different OBVs showing that the spikes are formed by an N-terminal extension of the fusion glycoprotein Gc. Using Schmallenberg virus, a recently emerged OBV, we also show that the projecting spike is the major target of the neutralizing antibody response, and provide X-ray structures in complex with two protecting antibodies. We further show that immunization of mice with the spike domains elicits virtually sterilizing immunity, providing fundamental knowledge essential in the preparation for potential newly emerging OBV zoonoses., Orthobunyaviruses (OBVs) cause severe disease in humans and farm animals, but the molecular basis for infection is not fully understood. Here, the authors present crystal structures of free and antibody-bound OBV envelope glycoproteins and show that their domains enable efficient immunization in a mouse model.
Published: 2019

13. Proteasomal degradation of NOD2 by NLRP12 in monocytes promotes bacterial tolerance and colonization by enteropathogens

Author: Normand, Sylvain, Waldschmitt, Nadine, Neerincx, Andreas, Martinez-Torres, Ruben Julio, Chauvin, Camille, Couturier-Maillard, Aurelie, Boulard, Olivier, Cobret, Laetitia, Awad, Fawaz, Huot, Ludovic, Ribeiro-Ribeiro, Andre, Lautz, Katja, Ruez, Richard, Delacre, Myriam, Bondu, Clovis, Guilliams, Martin, Scott, Charlotte, Segal, Anthony, Amselem, Serge, Hot, David, Karabina, Sonia, Bohn, Erwin, Ryffel, Bernhard, Poulin, Lionel F, Kufer, Thomas A, Chamaillard, Mathias, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM), University of Cambridge [UK] (CAM), University College of London [London] (UCL), Sorbonne Université - Faculté de Médecine (SU FM), Sorbonne Université (SU), Maladies génétiques d'expression pédiatrique (U933), Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), University of Cologne, Universiteit Gent = Ghent University (UGENT), CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Eberhard Karls Universität Tübingen = Eberhard Karls University of Tuebingen, Immunologie et Neurogénétique Expérimentales et Moléculaires (INEM), Université d'Orléans (UO)-Centre National de la Recherche Scientifique (CNRS), Université d'Orléans (UO), University of Hohenheim, We thank Yvonne Postma for technical assistance., Centre d’Infection et d’Immunité de Lille (CIIL) - U1019 - UMR 8204 (CIIL), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Technische Universität München [München] (TUM), Physiopathologie des maladies génétiques d'expression pédiatrique, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire Trafic, Signalisation et Ciblage Intracellulaires, Institut Curie, Centre d'Immunologie de Marseille - Luminy (CIML), Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Unit of Immunoregulation and Mucosal Immunology [Ghent, Belgium], VIB Inflammation Research Center [Ghent, Belgium], UCL, Dept Med, Eberhard Karls Universität Tübingen, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Physiopathologie des maladies génétiques d'expression pédiatrique (UMRS_933), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Universiteit Gent = Ghent University [Belgium] (UGENT), Gestionnaire, Hal Sorbonne Université, Maladies génétiques d'expression pédiatrique [CHU Trousseau] (Inserm U933), and UF de Génétique moléculaire [CHU Trousseau]
Subjects: Male, MONARCH-1, [SDV]Life Sciences [q-bio], Nod2 Signaling Adaptor Protein, ATG16L1, PROTEIN, MESH: Mice, Knockout, ACTIVATION, Mice, ATTENUATES COLON INFLAMMATION, MESH: Intracellular Signaling Peptides and Proteins/genetics, Medicine and Health Sciences, MESH: Animals, lcsh:Science, MESH: Inflammation/immunology, ComputingMilieux_MISCELLANEOUS, MESH: Enterobacteriaceae Infections/immunology, MESH: HSP90 Heat-Shock Proteins/metabolism, Mice, Knockout, CITROBACTER-RODENTIUM, Enterobacteriaceae Infections, Intracellular Signaling Peptides and Proteins, NF-kappa B, MESH: HEK293 Cells, MESH: Immune Tolerance/immunology, [SDV.IMM]Life Sciences [q-bio]/Immunology, SENSORS NOD1, MESH: NF-kappa B/metabolism, Acetylmuramyl-Alanyl-Isoglutamine, MESH: Citrobacter rodentium/immunology, MESH: Nod2 Signaling Adaptor Protein/metabolism, EXPRESSION, MESH: Intracellular Signaling Peptides and Proteins/metabolism, [SDV.IMM] Life Sciences [q-bio]/Immunology, Science, MESH: Acetylmuramyl-Alanyl-Isoglutamine/metabolism, DENDRITIC CELLS, Article, Cell Line, MESH: Bacterial Capsules/metabolism, Immune Tolerance, MESH: Enterobacteriaceae Infections/microbiology, Animals, Humans, HSP90 Heat-Shock Proteins, MESH: Mice, Bacterial Capsules, Inflammation, MESH: Humans, MUTATIONS, MESH: Gastrointestinal Microbiome/immunology, Ubiquitination, Biology and Life Sciences, MESH: Inflammation/microbiology, MESH: Male, Gastrointestinal Microbiome, MESH: Cell Line, HEK293 Cells, Citrobacter rodentium, MESH: Ubiquitination, lcsh:Q
Abstract: Mutations in the nucleotide-binding oligomerization domain protein 12 (NLRP12) cause recurrent episodes of serosal inflammation. Here we show that NLRP12 efficiently sequesters HSP90 and promotes K48-linked ubiquitination and degradation of NOD2 in response to bacterial muramyl dipeptide (MDP). This interaction is mediated by the linker-region proximal to the nucleotide-binding domain of NLRP12. Consequently, the disease-causing NLRP12 R284X mutation fails to repress MDP-induced NF-κB and subsequent activity of the JAK/STAT signaling pathway. While NLRP12 deficiency renders septic mice highly susceptible towards MDP, a sustained sensing of MDP through NOD2 is observed among monocytes lacking NLRP12. This loss of tolerance in monocytes results in greater colonization resistance towards Citrobacter rodentium. Our data show that this is a consequence of NOD2-dependent accumulation of inflammatory mononuclear cells that correlates with induction of interferon-stimulated genes. Our study unveils a relevant process of tolerance towards the gut microbiota that is exploited by an attaching/effacing enteric pathogen., Mutations in nucleotide-binding oligomerization domain protein 12 (NLRP12) are known to effect inflammatory processes. Here the authors show that NLRP12-mediated proteasomal degradation of NOD2 in monocytes promotes bacterial tolerance and colonisation in a model of enteric infection.
Published: 2018

14. Thermogenic recruitment of brown and brite/beige adipose tissues is not obligatorily associated with macrophage accretion or attrition

Author: Nathalie Boulet, Jan Nedergaard, Barbara Cannon, Ineke H.N. Luijten, boulet, nathalie, and Stockholm University
Subjects: 0301 basic medicine, Male, obesity, Physiology, Endocrinology, Diabetes and Metabolism, [SDV]Life Sciences [q-bio], Adipose tissue, MESH: Adipose Tissue, Beige, MESH: Mice, Knockout, Mice, MESH: Uncoupling Protein 1, 0302 clinical medicine, Adipose Tissue, Brown, Brown adipose tissue, Macrophage, MESH: Obesity, MESH: Thermogenesis, MESH: Animals, Biological sciences, Uncoupling Protein 1, Mice, Knockout, Thermogenesis, MESH: Gene Expression Regulation, macrophages, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, Research Article, medicine.medical_specialty, Biology, MESH: Adipose Tissue, Brown, 03 medical and health sciences, MESH: Diet, MESH: Mice, Inbred C57BL, Physiology (medical), Internal medicine, medicine, Animals, MESH: Mice, MESH: Macrophages, brown adipose tissue, Adipose Tissue, Beige, cold, MESH: Male, Diet, Mice, Inbred C57BL, 030104 developmental biology, Endocrinology, Gene Expression Regulation, Receptors, Adrenergic, beta-2, MESH: Receptors, Adrenergic, beta-1, Receptors, Adrenergic, beta-1, MESH: Receptors, Adrenergic, beta-2, 030217 neurology & neurosurgery
Abstract: Cold- and diet-induced recruitment of brown adipose tissue (BAT) and the browning of white adipose tissue (WAT) are dynamic processes, and the recruited state attained is a state of dynamic equilibrium, demanding continuous stimulation to be maintained. An involvement of macrophages, classical proinflammatory (M1) or alternatively activated anti-inflammatory (M2), is presently discussed as being an integral part of these processes. If these macrophages play a mediatory role in the recruitment process, such an involvement would have to be maintained in the recruited state. We have, therefore, investigated whether the recruited state of these tissues is associated with macrophage accretion or attrition. We found no correlation (positive or negative) between total UCP1 mRNA levels (as a measure of recruitment) and proinflammatory macrophages in any adipose depot. We found that in young chow-fed mice, cold-induced recruitment correlated with accretion of anti-inflammatory macrophages; however, such a correlation was not seen when cold-induced recruitment was studied in diet-induced obese mice. Furthermore, the anti-inflammatory macrophage accretion was mediated via β1/β2-adrenergic receptors; yet, in their absence, and thus in the absence of macrophage accretion, recruitment proceeded normally. We thus conclude that the classical recruited state in BAT and inguinal (brite/beige) WAT is not paralleled by macrophage accretion or attrition. Our results make mediatory roles for macrophages in the recruitment process less likely. NEW & NOTEWORTHY A regulatory or mediatory role—positive or negative—for macrophages in the recruitment of brown adipose tissue is presently discussed. As the recruited state in the tissue is a dynamic process, maintenance of the recruited state would need persistent alterations in macrophage complement. Contrary to this expectation, we demonstrate here an absence of alterations in macrophage complement in thermogenically recruited brown—or brite/beige—adipose tissues. Macrophage regulation of thermogenic capacity is thus less likely.
Published: 2021

15. The activation trajectory of plasmacytoid dendritic cells in vivo during a viral infection

Author: Jean-Luc Davignon, Karima Naciri, Rabie Chelbi, Gilles Bessou, Chuang Dong, Nils Collinet, Michael Valente, Geoffray Brelurut, Pierre Milpied, Abdenour Abbas, Morgane Thomas-Chollier, Noudjoud Attaf, Thien-Phong Vu Manh, Inaki Cervera-Marzal, Alexandra-Chloé Villani, Benjamin Rauwel, Denis Thieffry, Marc Dalod, Elena Tomasello, Centre d'Immunologie de Marseille - Luminy (CIML), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie de l'ENS Paris (IBENS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Biologie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre de Physiopathologie Toulouse Purpan (CPTP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, USA, Partenaires INRAE, Center for Immunology and Inflammatory Diseases [Boston, MA, USA], Institut de biologie de l'ENS Paris (UMR 8197/1024) (IBENS), Département de Biologie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and European Project: 281225,EC:FP7:ERC,ERC-2011-StG_20101109,SYSTEMSDENDRITIC(2012)
Subjects: CD4-Positive T-Lymphocytes, 0301 basic medicine, MESH: CD4-Positive T-Lymphocytes / immunology, MESH: Signal Transduction, Muromegalovirus, [SDV]Life Sciences [q-bio], Leukemia inhibitory factor receptor, Lymphocyte Activation, MESH: Mice, Knockout, Mice, 0302 clinical medicine, Single-cell analysis, Interferon, MESH: Sequence Analysis, RNA, Immunology and Allergy, MESH: Microscopy, Confocal, MESH: Animals, MESH: Interferon Type I / metabolism, MESH: Tumor Necrosis Factor-alpha / metabolism, Cells, Cultured, ComputingMilieux_MISCELLANEOUS, Mice, Knockout, Mice, Inbred BALB C, Microscopy, Confocal, medicine.diagnostic_test, hemic and immune systems, Herpesviridae Infections, 3. Good health, Cell biology, medicine.anatomical_structure, Plasmacytoid dendritic cells, Interferon Type I, MESH: Herpesviridae Infections / immunology, Tumor necrosis factor alpha, Single-Cell Analysis, MESH: Dendritic Cells / immunology, Signal Transduction, medicine.drug, MESH: Cells, Cultured, T cell, Immunology, MESH: Mice, Inbred BALB C, Biology, Article, Flow cytometry, 03 medical and health sciences, Gene expression analysis, Downregulation and upregulation, In vivo, MESH: Mice, Inbred C57BL, medicine, Animals, MESH: Lymphocyte Activation, Sequence Analysis, RNA, Tumor Necrosis Factor-alpha, Dendritic Cells, Mice, Inbred C57BL, Confocal microscopy, 030104 developmental biology, MESH: Muromegalovirus / physiology, 030215 immunology, MESH: Single-Cell Analysis
Abstract: International audience; Plasmacytoid dendritic cells (pDCs) are a major source of type I interferon (IFN-I). What other functions pDCs exert in vivo during viral infections is controversial, and more studies are needed to understand their orchestration. In the present study, we characterize in depth and link pDC activation states in animals infected by mouse cytomegalovirus by combining Ifnb1 reporter mice with flow cytometry, single-cell RNA sequencing, confocal microscopy and a cognate CD4 T cell activation assay. We show that IFN-I production and T cell activation were performed by the same pDC, but these occurred sequentially in time and in different micro-anatomical locations. In addition, we show that pDC commitment to IFN-I production was marked early on by their downregulation of leukemia inhibitory factor receptor and was promoted by cell-intrinsic tumor necrosis factor signaling. We propose a new model for how individual pDCs are endowed to exert different functions in vivo during a viral infection, in a manner tightly orchestrated in time and space.
Published: 2020

16. The HSP90/R2TP assembly chaperone promotes cell proliferation in the intestinal epithelium

Author: Marina Ferrand, Edouard Bertrand, Bénédicte Lemmers, Claire Abeza, Francina Langa, Conception Paul, Céline Verheggen, Valérie Pinet, Dominique Helmlinger, Bérengère Pradet-Balade, Rossano Lattanzio, Ciro Longobardi, Michael Hahne, Philippe Jay, François Gerbe, Monica Gabola, Julie Bremond, Chloé Maurizy, Nicola Tinari, Institut de Génétique Moléculaire de Montpellier (IGMM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), Institut de Génomique Fonctionnelle (IGF), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Institut de génétique humaine (IGH), Università degli studi 'G. d'Annunzio' Chieti-Pescara [Chieti-Pescara] (Ud'A), Centre de recherche en Biologie cellulaire de Montpellier (CRBM), The work was supported by La Ligue Nationale Contre le Cancer (équipes labelisées to E.B., P.J.) and the INCa grants PLBIO 2016-161 to D.H., E.B., M.H., B.P.B., and PLBIO 2018-158 to P.J. C.M. and C.A. had fellowships from la Ligue Nationale Contre le Cancer., Guerineau, Nathalie C., Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Institut Pasteur [Paris], Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche en Biologie Cellulaire (CRBM), and Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1)
Subjects: Cellular differentiation, General Physics and Astronomy, MESH: Mice, Knockout, 0302 clinical medicine, Chaperones, MESH: Microscopy, Confocal, MESH: Animals, Intestinal Mucosa, Tissue homeostasis, Cells, Cultured, Mice, Knockout, 0303 health sciences, Multidisciplinary, Microscopy, Confocal, biology, Chemistry, Stem Cells, Intestinal stem cells, Intestinal epithelium, 3. Good health, Cell biology, MESH: Epithelial Cells, 030220 oncology & carcinogenesis, MESH: Intestinal Mucosa, Stem cell, MESH: Molecular Chaperones, MESH: Cells, Cultured, Protein Binding, MESH: Mice, Transgenic, Science, Mice, Transgenic, MESH: Stem Cells, Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, MESH: Mice, Inbred C57BL, MESH: Cell Proliferation, MESH: HSP90 Heat-Shock Proteins, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Protein Binding, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, HSP90 Heat-Shock Proteins, Progenitor cell, PI3K/AKT/mTOR pathway, 030304 developmental biology, Cell Proliferation, MESH: Humans, Cell growth, Epithelial Cells, General Chemistry, Mice, Inbred C57BL, Chaperone (protein), biology.protein, Molecular Chaperones
Abstract: The R2TP chaperone cooperates with HSP90 to integrate newly synthesized proteins into multi-subunit complexes, yet its role in tissue homeostasis is unknown. Here, we generated conditional, inducible knock-out mice for Rpap3 to inactivate this core component of R2TP in the intestinal epithelium. In adult mice, Rpap3 invalidation caused destruction of the small intestinal epithelium and death within 10 days. Levels of R2TP substrates decreased, with strong effects on mTOR, ATM and ATR. Proliferative stem cells and progenitors deficient for Rpap3 failed to import RNA polymerase II into the nucleus and they induced p53, cell cycle arrest and apoptosis. Post-mitotic, differentiated cells did not display these alterations, suggesting that R2TP clients are preferentially built in actively proliferating cells. In addition, high RPAP3 levels in colorectal tumors from patients correlate with bad prognosis. Here, we show that, in the intestine, the R2TP chaperone plays essential roles in normal and tumoral proliferation., RPAP3 is a subunit of the R2TP complex, a co-chaperone of HSP90, with substrate proteins involved in transcription, ribosome biogenesis, DNA repair and cell growth. Here the authors report that deletion of Rpap3 abrogates cell proliferation and homeostasis in mouse intestine, partly through destabilization of PI3K-like kinases, while elevated RPAP3 levels in colorectal tumors are associated with poor prognosis.
Published: 2020

17. Metformin lowers glucose 6-phosphate in hepatocytes by activation of glycolysis downstream of glucose phosphorylation

Author: Ahmed Alshawi, Loranne Agius, Marc Foretz, Benoit Viollet, Silvia Marin, Natasha S. Adam-Primus, Shruti S. Chachra, Ziad H. Al-Oanzi, Tabassum Moonira, Marta Cascante, Catherine Arden, Brian E. Ford, Institut Cochin (IC UM3 (UMR 8104 / U1016)), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)
Subjects: 0301 basic medicine, Male, Phosphofructokinase-1, Hypoglucemic agents, AMP-Activated Protein Kinases, MESH: Metformin, MESH: Mice, Knockout, Biochemistry, MESH: Hepatocytes, chemistry.chemical_compound, Mice, Adenosine Triphosphate, MESH: Adenosine Triphosphate, Malalties cròniques, MESH: Animals, Glycolysis, MESH: AMP-Activated Protein Kinases, Phosphorylation, Antidiabètics, Mice, Knockout, Chemistry, MESH: Gluconeogenesis, MESH: Glycerolphosphate Dehydrogenase, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, glycolysis, MESH: Glucose-6-Phosphate, Metformin, 3. Good health, MESH: Glucose, phosphofructokinase, MESH: Glycolysis, Dihydroxyacetone, medicine.drug, Phosphofructokinase, medicine.medical_specialty, MESH: Rats, Allosteric regulation, Glucose-6-Phosphate, Glycerolphosphate Dehydrogenase, liver, 03 medical and health sciences, MESH: Phosphofructokinase-1, MESH: Mice, Inbred C57BL, Internal medicine, Rotenone, medicine, hepatocyte, Animals, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Rats, Wistar, MESH: Mice, Molecular Biology, glucose 6-phosphate, MESH: Dihydroxyacetone, MESH: Phosphorylation, 030102 biochemistry & molecular biology, Gluconeogenesis, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Fructose, MESH: Rats, Wistar, Cell Biology, Metabolism, MESH: Male, Rats, Mice, Inbred C57BL, 030104 developmental biology, Endocrinology, Glucose, Glucose 6-phosphate, Chronic diseases, Hepatocytes, MESH: Rotenone
Abstract: International audience; The chronic effects of metformin on liver gluconeogenesisinvolve repression of theG6pcgene, which is regulated by thecarbohydrate-response element–binding protein throughraised cellular intermediates of glucose metabolism. In thisstudy we determined the candidate mechanisms by which met-formin lowers glucose 6-phosphate (G6P) in mouse and rathepatocytes challenged with high glucose or gluconeogenic pre-cursors. Cell metformin loads in the therapeutic range loweredcell G6P but not ATP and decreasedG6pcmRNA at high glu-cose. The G6P lowering by metformin was mimicked by a com-plex 1 inhibitor (rotenone) and an uncoupler (dinitrophenol)and by overexpression of mGPDH, which lowers glycerol3-phosphate and G6P and also mimics theG6pcrepression bymetformin. In contrast, direct allosteric activators of AMPK(A-769662, 991, and C-13) had opposite effects from metforminon glycolysis, gluconeogenesis, and cell G6P. The G6P loweringby metformin, which also occurs in hepatocytes from AMPKknockout mice, is best explained by allosteric regulation ofphosphofructokinase-1 and/or fructose bisphosphatase-1, assupported by increased metabolism of [3-3H]glucose relative to[2-3H]glucose; by an increase in the lactate m2/m1 isotopologratio from [1,2-13C2]glucose; by lowering of glycerol 3-phos-phate an allosteric inhibitor of phosphofructokinase-1; and bymarked G6P elevation by selective inhibition of phosphofruc-tokinase-1; but not by a more reduced cytoplasmic NADH/NADredox state. We conclude that therapeutically relevant doses of metformin lower G6P in hepatocytes challenged with high glu-cose by stimulation of glycolysis by an AMP-activated proteinkinase–independent mechanism through changes in allostericeffectors of phosphofructokinase-1 and fructose bisphospha-tase-1, including AMP, Pi, and glycerol 3-phosphate
Published: 2020

18. TGR5-dependent hepatoprotection through the regulation of biliary epithelium barrier function

Author: Jose Ursic-Bedoya, Isabelle Doignon, Hayat Simerabet, Dominique Rainteau, Doris Cassio, Nicolas Kahale, Anne Davit-Spraul, Thierry Tordjmann, Grégory Merlen, Lydie Humbert, Catherine Guettier, Julien Gautherot, Valeska Bidault-Jourdainne, Christoph Ullmer, Isabelle Garcin, Noémie Péan, Klaus Ebnet, Zahra Tanfin, Intéractions cellulaires et physiopathologie hépathique (Orsay, Essonne) UMRS 1174 (ICPH ), Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris-Sud - Paris 11 - Faculté de médecine (UP11 UFR Médecine), Université Paris-Sud - Paris 11 (UP11), Service de Biochimie [AP-HP Hôpital Bicêtre], AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre), Service d’Anatomopathologie [Le Kremlin-Bicêtre], ER_7, Université Pierre et Marie Curie - Paris 6 (UPMC), Microorganismes, Molécules Bioactives et Physiopathologie Intestinale (LBM-E4), Institut National de la Santé et de la Recherche Médicale (INSERM)-Laboratoire des biomolécules (LBM UMR 7203), Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Département de Chimie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Chimie Moléculaire de Paris Centre (FR 2769), Institut de Chimie du CNRS (INC)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Microorganismes et physiopathologie intestinale (ERL INSERM U1157 - CNRS UMR 7203), Laboratoire des biomolécules (LBM UMR 7203), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC), Zentrum für Molekularbiologie der Entzündung - Center for Molecular Biology of Inflammation [Münster, Germany] (ZMBE), Westfälische Wilhelms-Universität Münster (WWU), F. Hoffmann-La Roche [Basel], Chimie Moléculaire de Paris Centre (FR 2769), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Chimie Moléculaire de Paris Centre (FR 2769), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Westfälische Wilhelms-Universität Münster = University of Münster (WWU), and Salvy-Córdoba, Nathalie
Subjects: Biliary epithelium, MESH: Signal Transduction, Stimulation, MESH: Receptors, G-Protein-Coupled, MESH: Mice, Knockout, Epithelium, Receptors, G-Protein-Coupled, MESH: Isonipecotic Acids, 0302 clinical medicine, MESH: Biliary Tract, Oximes, Electric Impedance, Bile, MESH: Animals, Phosphorylation, Biliary Tract, Cells, Cultured, Barrier function, MESH: Receptors, Cell Surface, Mice, Knockout, Liver injury, 0303 health sciences, Bile acid, Tight junction, Chemistry, Gastroenterology, MESH: Oximes, G protein-coupled bile acid receptor, Paracellular transport, MESH: Permeability, Cholestaticliver diseases, MESH: Cell Adhesion Molecules, 030211 gastroenterology & hepatology, Signal Transduction, MESH: Cells, Cultured, medicine.drug_class, MESH: Tight Junction Proteins, Receptors, Cell Surface, Cholestasis, Intrahepatic, MESH: Bile Acids and Salts, Permeability, Bile Acids and Salts, 03 medical and health sciences, Isonipecotic Acids, In vivo, MESH: Mice, Inbred C57BL, medicine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Electric Impedance, MESH: Bile, 030304 developmental biology, Tight Junction Proteins, MESH: Phosphorylation, MESH: Cholestasis, Intrahepatic, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, medicine.disease, Molecular biology, [SDV.MHEP.HEG] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, Mice, Inbred C57BL, MESH: Epithelium, Cell Adhesion Molecules
Abstract: ObjectiveWe explored the hypothesis that TGR5, the bile acid (BA) G-protein-coupled receptor highly expressed in biliary epithelial cells, protects the liver against BA overload through the regulation of biliary epithelium permeability.DesignExperiments were performed under basal and TGR5 agonist treatment. In vitro transepithelial electric resistance (TER) and FITC-dextran diffusion were measured in different cell lines. In vivo FITC-dextran was injected in the gallbladder (GB) lumen and traced in plasma. Tight junction proteins and TGR5-induced signalling were investigated in vitro and in vivo (wild-type [WT] and TGR5-KO livers and GB). WT and TGR5-KO mice were submitted to bile duct ligation or alpha-naphtylisothiocyanate intoxication under vehicle or TGR5 agonist treatment, and liver injury was studied.ResultsIn vitro TGR5 stimulation increased TER and reduced paracellular permeability for dextran. In vivo dextran diffusion after GB injection was increased in TGR5-knock-out (KO) as compared with WT mice and decreased on TGR5 stimulation. In TGR5-KO bile ducts and GB, junctional adhesion molecule A (JAM-A) was hypophosphorylated and selectively downregulated among TJP analysed. TGR5 stimulation induced JAM-A phosphorylation and stabilisation both in vitro and in vivo, associated with protein kinase C-ζ activation. TGR5 agonist-induced TER increase as well as JAM-A protein stabilisation was dependent on JAM-A Ser285 phosphorylation. TGR5 agonist-treated mice were protected from cholestasis-induced liver injury, and this protection was significantly impaired in JAM-A-KO mice.ConclusionThe BA receptor TGR5 regulates biliary epithelial barrier function in vitro and in vivo through an impact on JAM-A expression and phosphorylation, thereby protecting liver parenchyma against bile leakage.
Published: 2019

19. A genetic variant controls interferon-β gene expression in human myeloid cells by preventing C/EBP-β binding on a conserved enhancer

Author: Didier Busso, Lluis Quintana-Murci, Paul-Henri Romeo, Juliette Hamroune, Anaïs Assouvie, Germain Rousselet, Maxime Rotival, Laboratoire de Recherche sur la Réparation et la Transcription dans les Cellules Souches (LRTS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Génétique Evolutive Humaine - Human Evolutionary Genetics, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), U 1016/ UMR 8104 Institut Cochin, Plate-Forme Séquençage et Génomique (Genom'IC), Institut National de la Santé et de la Recherche Médicale (INSERM), Stabilité génétique, cellules souches et radiations (SGCSR (U_1274 / UMR_E_008)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Université de Paris (UP), Chaire Génomique humaine et évolution, Collège de France (CdF (institution)), La Ligue contre le cancer, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Université Paris Cité (UPCité), Collège de France - Chaire Génomique humaine et évolution, and Quintana-Murci, lluis
Subjects: Lipopolysaccharides, Cancer Research, Myeloid, Single Nucleotide Polymorphisms, Gene Expression, QH426-470, MESH: Mice, Knockout, Biochemistry, Monocytes, Mice, White Blood Cells, 0302 clinical medicine, Transcription (biology), Animal Cells, Gene expression, Medicine and Health Sciences, MESH: Animals, Myeloid Cells, Promoter Regions, Genetic, Genetics (clinical), Cells, Cultured, Mice, Knockout, 0303 health sciences, Mammalian Genomics, MESH: Polymorphism, Single Nucleotide, MESH: Transcription Factors, Genomics, MESH: Gene Expression Regulation, 3. Good health, Enzymes, medicine.anatomical_structure, Enhancer Elements, Genetic, Cellular Types, Oxidoreductases, Luciferase, MESH: Cells, Cultured, Research Article, MESH: Blood Buffy Coat, Immune Cells, Primary Cell Culture, Quantitative Trait Loci, Immunology, Bone Marrow Cells, [SDV.GEN.GH] Life Sciences [q-bio]/Genetics/Human genetics, Biology, Polymorphism, Single Nucleotide, MESH: CCAAT-Enhancer-Binding Protein-beta, MESH: Primary Cell Culture, 03 medical and health sciences, MESH: Promoter Regions, Genetic, DNA-binding proteins, medicine, Genetics, Animals, Humans, Point Mutation, Gene Regulation, Allele, Enhancer, Molecular Biology, Transcription factor, Gene, MESH: Mice, Ecology, Evolution, Behavior and Systematics, Alleles, 030304 developmental biology, MESH: Point Mutation, MESH: Humans, Blood Cells, MESH: Interferon-beta, MESH: Alleles, CCAAT-Enhancer-Binding Protein-beta, Macrophages, Biology and Life Sciences, Proteins, Interferon-beta, Cell Biology, TRIM33, Molecular biology, MESH: Myeloid Cells, MESH: Quantitative Trait Loci, Regulatory Proteins, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, Gene Expression Regulation, Animal Genomics, Blood Buffy Coat, Enzymology, MESH: Enhancer Elements, Genetic, MESH: Lipopolysaccharides, 030217 neurology & neurosurgery, Transcription Factors
Abstract: Interferon β (IFN-β) is a cytokine that induces a global antiviral proteome, and regulates the adaptive immune response to infections and tumors. Its effects strongly depend on its level and timing of expression. Therefore, the transcription of its coding gene IFNB1 is strictly controlled. We have previously shown that in mice, the TRIM33 protein restrains Ifnb1 transcription in activated myeloid cells through an upstream inhibitory sequence called ICE. Here, we show that the deregulation of Ifnb1 expression observed in murine Trim33-/- macrophages correlates with abnormal looping of both ICE and the Ifnb1 gene to a 100 kb downstream region overlapping the Ptplad2/Hacd4 gene. This region is a predicted myeloid super-enhancer in which we could characterize 3 myeloid-specific active enhancers, one of which (E5) increases the response of the Ifnb1 promoter to activation. In humans, the orthologous region contains several single nucleotide polymorphisms (SNPs) known to be associated with decreased expression of IFNB1 in activated monocytes, and loops to the IFNB1 gene. The strongest association is found for the rs12553564 SNP, located in the E5 orthologous region. The minor allele of rs12553564 disrupts a conserved C/EBP-β binding motif, prevents binding of C/EBP-β, and abolishes the activation-induced enhancer activity of E5. Altogether, these results establish a link between a genetic variant preventing binding of a transcription factor and a higher order phenotype, and suggest that the frequent minor allele (around 30% worldwide) might be associated with phenotypes regulated by IFN-β expression in myeloid cells., Author summary Genome-wide association studies identify multiple genetic variants associated with higher order phenotypes. Pinpointing the causative variant and understanding its molecular mode of action is a complex task. Using a murine model of interferon-β transcriptional deregulation, we characterize a super-enhancer controlling Ifnb1 expression in myeloid cells. The most active enhancer of this locus is conserved in humans, but presents a frequent variant found in around 30% of the population worldwide. This variant prevents binding of the C/EBP-β transcription factor, and is associated with decreased expression of IFNB1 in activated monocytes. When mimicked in the murine enhancer, it abolishes its inducible enhancer activity. Our results describe the molecular link between a point mutation and a cellular phenotype that could influence clinical situations.
Published: 2019

20. Mouse models for the study of fate and function of innate lymphoid cells

Author: François Dejardin, Jasna Medvedovic, Emelyne Lécuyer, Tegest Aychek, Gérard Eberl, Sascha Cording, Microenvironnement et Immunité - Microenvironment and Immunity, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratory of Intestinal Immunity (Equipe Inserm U1163), Imagine - Institut des maladies génétiques (IMAGINE - U1163), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Diderot, Sorbonne Paris Cité, Paris, France, Université Paris Diderot - Paris 7 (UPD7), Fondation pour la Recherche MédicaleCrohn's and Colitis Foundation of AmericaInstitut Pasteur, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), and HUGOT, Bérengère
Subjects: MESH: Killer Cells, Natural, 0301 basic medicine, Cell type, [SDV.IMM] Life Sciences [q-bio]/Immunology, Lymphoid Tissue, Immunology, Gata3, Innate lymphoid cells, Biology, Mouse models, T-bet, MESH: Mice, Knockout, Mice, 03 medical and health sciences, 0302 clinical medicine, RAR-related orphan receptor gamma, Animals, Immunology and Allergy, Cell Lineage, MESH: Animals, Lymphocytes, MESH: T-Lymphocytes, Helper-Inducer, MESH: Mice, Mice, Knockout, Innate lymphoid cell, GATA3, T-Lymphocytes, Helper-Inducer, MESH: Cell Lineage, Acquired immune system, Phenotype, Immunity, Innate, Cell biology, Killer Cells, Natural, 030104 developmental biology, Lymphatic system, MESH: Models, Animal, Models, Animal, MESH: Lymphoid Tissue, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Immunity, Innate, MESH: Lymphocytes, RORγt, Function (biology), 030215 immunology
Abstract: International audience; Natural killer (NK) cells and lymphoid tissue inducer (LTi) cells were discovered more than 40 and 20 years ago, respectively. These two cell types were initially studied for their unique functions in the elimination of infected or transformed cells, and in the development of lymphoid tissues. It took an additional 10 years to realize that NK cells and LTi cells were members of a larger family of innate lymphoid cells (ILCs), whose phenotypes and functions mirror those of T cells. Many mouse models have since been developed to identify and isolate ILCs, map their developmental pathways and characterize their functions. Because of the similarity between ILCs and T cells, this exploration remains a challenge. In spite of this, a broad range of mouse models available to researchers has lead to significant progress in untangling the unique roles of ILCs early in defense, regulation of adaptive immunity and homeostasis. Here, we review these mouse models, and discuss their strengths and limitations.
Published: 2018

21. α1AMPK deletion in myelomonocytic cells induces a pro-inflammatory phenotype and enhances angiotensin II-induced vascular dysfunction

Author: Marc Foretz, Moritz Brandt, Eberhard Schulz, Philip Wenzel, Matthias Oelze, John F. Keaney, Swenja Kröller-Schön, Thomas Münzel, Benoit Viollet, Thomas Jansen, Andreas Daiber, Sebastian Steven, Jeremy Lagrange, Miroslava Kvandova, Sanela Kalinovic, Tanja Schönfelder, Johannes Gutenberg - Universität Mainz (JGU), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Paris 5 (UPD5), University of Massachusetts Medical School [Worcester] (UMASS), University of Massachusetts System (UMASS), and Johannes Gutenberg - University of Mainz (JGU)
Subjects: MESH: Signal Transduction, 0301 basic medicine, CCR2, Physiology, medicine.medical_treatment, MESH: Aortic Diseases, AMP-Activated Protein Kinases, 030204 cardiovascular system & hematology, MESH: Mice, Knockout, 0302 clinical medicine, MESH: Animals, MESH: AMP-Activated Protein Kinases, Aorta, Cells, Cultured, Mice, Knockout, MESH: Cytokines, MESH: Oxidative Stress, biology, Chemistry, Angiotensin II, MESH: Genetic Predisposition to Disease, MESH: Aorta, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Respiratory burst, Vasodilation, Phenotype, Cytokine, Cytokines, Tumor necrosis factor alpha, MESH: Angiotensin II, Inflammation Mediators, medicine.symptom, Cardiology and Cardiovascular Medicine, Signal Transduction, MESH: Cells, Cultured, MESH: Inflammation Mediators, Aortic Diseases, Inflammation, MESH: Phenotype, MESH: Vasodilation, 03 medical and health sciences, Immune system, MESH: Mice, Inbred C57BL, Physiology (medical), medicine, Animals, Genetic Predisposition to Disease, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Interleukin 6, Macrophages, MESH: Macrophages, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Mice, Inbred C57BL, Disease Models, Animal, Oxidative Stress, 030104 developmental biology, MESH: Gene Deletion, Cancer research, biology.protein, MESH: Disease Models, Animal, Gene Deletion
Abstract: Aims Immune cell function involves energy-dependent processes including growth, proliferation, and cytokine production. Since the AMP-activated protein kinase (AMPK) is a crucial regulator of intracellular energy homeostasis, its expression and activity may also affect innate and adaptive immune cell responses. Therefore, we aimed to investigate the consequences of α1AMPK deletion in myelomonocytic cells on vascular function, inflammation, and hypertension during chronic angiotensin II (ATII) treatment. Methods and results We generated a mouse strain with α1AMPK deletion in lysozyme M+ myelomonocytic cells. Compared to controls, chronic ATII infusion (1 mg/kg/day for 7 days) lead to increased vascular oxidative stress and aggravated endothelial dysfunction in LysM-Cre+ x α1AMPKfl/fl mice. This was accompanied by an increased aortic infiltration of CD11b+F4/80+ macrophages and enhanced pro-inflammatory cytokine release (tumour necrosis factor-alpha, interferon-gamma, and interleukin-6). Mechanistically, we found that increased expression of C-C chemokine receptor 2 (CCR2) in α1AMPK deficient myelomonocytic cells facilitated their recruitment to the vascular wall. In addition, expression of the ATII receptor type 1a and the oxidative burst was increased in these cells, indicating an increased susceptibility towards pro-oxidant stimuli. Conclusions In summary, α1AMPK deletion in myelomonocytic cells aggravates vascular oxidative stress and dysfunction by enhancing their recruitment to the vascular wall and increasing their susceptibility towards pro-oxidant stimuli. Our observations suggest that metabolic control in myelomonocytic cells has profound implications for their inflammatory phenotype and may trigger the development of vascular disease.
Published: 2018

22. The Wnt/Ca 2+ pathway is involved in interneuronal communication mediated by tunneling nanotubes

Author: Gonzalo Córdova, Jessica Y Vargas, Frida Loria, Chiara Zurzolo, Masato Hasegawa, Geeske M. van Woerden, Yuan-Ju Wu, Sebastien Bellow, Takashi Nonaka, Capucine Trollet, Sylvie Syan, Trafic membranaire et Pathogénèse, Institut Pasteur [Paris], Institut de Myologie, Centre National de la Recherche Scientifique (CNRS)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Association française contre les myopathies (AFM-Téléthon)-Sorbonne Université (SU), Centre de recherche en Myologie – U974 SU-INSERM, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Tokyo Metropolitan University [Tokyo] (TMU), Bioaxial, Department of Neuroscience [Rotterdam, the Netherlands], Erasmus University Medical Center [Rotterdam] (Erasmus MC), Expertise Center for Neurodevelopmental Disorders [Rotterdam, the Netherlands] (ENCORE), This work was supported by the Agence Nationale de la Recherche [ANR‐16‐CE16‐0019‐01], Fondation pour la Recherche Médicale [FRM‐2016‐DEQ20160334896] and grants from LECMA‐Vaincre Alzheimer and France Alzheimer foundations to CZ. Funding from E‐Rare/ERANET (project SIRD) to CT. Grant‐in‐Aid for Scientific Research on Innovative Areas (Brain Protein Aging and Dementia Control) (JP26117005) from MEXT and Grant‐in‐Aid for Scientific Research on Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) (JP14533254) from AMED and JST CREST (JP18071300) to MH. Grant from Brain Science Foundation to TN. FL was recipient of Marie Skłodowska‐Curie individual fellowships and is funded through the Marie Skłodowska‐Curie Action COFUND 2015 (EU project 713366 – InterTalentum). Y‐JW was recipient of Pasteur‐Roux‐Cantarini fellowships., The authors are thankful to Dr. Seng Zhu and Shaarvari Bhat for their help in the acquisition of live imaging experiments., Institut Pasteur [Paris] (IP), ANR-16-CE16-0019,Neurotunn,Role des nanotubes membranaires dans la propagation d'agrégats protéiques impliqués dans les maladie neurodégénératives(2016), Department of Neuropathology and Cell Biology, Tokyo Metropolitan Institute of Medical Science, Biologie Cellulaire du Parasitisme, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Dipartimento di Biologia e Patologia Cellulare e Moleculare, Università degli studi di Napoli Federico II, Neurosciences, Centre de Recherche en Myologie, Vargas, J. Y., Loria, F., Wu, Y. -J., Cordova, G., Nonaka, T., Bellow, S., Syan, S., Hasegawa, M., van Woerden, G. M., Trollet, C., and Zurzolo, C.
Subjects: MESH: Signal Transduction, Wnt Protein, [SDV]Life Sciences [q-bio], MESH: Mice, Knockout, tunneling nanotubes, MESH: Nanotubes/chemistry, Mice, Cell Communication, MESH: Cell Communication, 0302 clinical medicine, MESH: Animals, MESH: Wnt Proteins/metabolism, Cells, Cultured, ComputingMilieux_MISCELLANEOUS, MESH: Neurons/cytology, Mice, Knockout, 0303 health sciences, CaMKII, General Neuroscience, Vesicle, Neurodegeneration, Wnt signaling pathway, musculoskeletal system, 3. Good health, Cell biology, intercellular communication, MESH: Calcium/metabolism, Intracellular, Signal Transduction, MESH: Cells, Cultured, Gene isoform, Wnt pathway, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH: Cell Membrane/metabolism, Biology, MESH: Calcium Signaling, MESH: Calcium-Calmodulin-Dependent Protein Kinase Type 2/physiology, General Biochemistry, Genetics and Molecular Biology, MESH: Neurons/physiology, 03 medical and health sciences, tunneling nanotube, α-synuclein, MESH: Mice, Inbred C57BL, Ca2+/calmodulin-dependent protein kinase, medicine, Calcium Signaling, MESH: Mice, Molecular Biology, Actin, 030304 developmental biology, General Immunology and Microbiology, Animal, Cell Membrane, MESH: Actins/metabolism, Neuron, medicine.disease, Mice, Inbred C57BL, Nanotube, Cell culture, Calcium, Calcium-Calmodulin-Dependent Protein Kinase Type 2, 030217 neurology & neurosurgery
Abstract: International audience; Tunneling nanotubes (TNTs) are actin-based transient tubular connections that allow direct communication between distant cells. TNTs play an important role in several physiological (development, immunity, and tissue regeneration) and pathological (cancer, neurodegeneration, and pathogens transmission) processes. Here, we report that the Wnt/Ca2+ pathway, an intracellular cascade that is involved in actin cytoskeleton remodeling, has a role in TNT formation and TNT-mediated transfer of cargoes. Specifically, we found that Ca2+ /calmodulin-dependent protein kinase II (CaMKII), a transducer of the Wnt/Ca2+ pathway, regulates TNTs in a neuronal cell line and in primary neurons. We identified the β isoform of CaMKII as a key molecule in modulating TNT formation and transfer, showing that this depends on the actin-binding activity of the protein. Finally, we found that the transfer of vesicles and aggregated α-synuclein between primary neurons can be regulated by the activation of the Wnt/Ca2+ pathway. Our findings suggest that Wnt/Ca2+ pathway could be a novel promising target for therapies designed to impair TNT-mediated propagation of pathogens.
Published: 2019

23. Predominant role of microglia in brain iron retention in Sanfilippo syndrome, a pediatric neurodegenerative disease

Author: Laurent Puy, Cathy Gomila, Jérôme Ausseil, Agnès Boullier, Zoubida Karim, Stéphanie Trudel, Christelle Lony, Sandrine Vitry, Thibaud Lefebvre, Walaa Darwiche, Vincent Puy, Université de Picardie Jules Verne (UPJV), North Hospital, Centre de recherche sur l'Inflammation (CRI (UMR_S_1149 / ERL_8252 / U1149)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7), Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Inserm, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre hospitalier universitaire d'Amiens (CHU Amiens-Picardie), CHU Amiens-Picardie, Mécanismes physiologiques et conséquences des calcifications cardiovasculaires: rôle des remodelages cardiovasculaires et osseux, Université de Picardie Jules Verne (UPJV)-Institut National de la Santé et de la Recherche Médicale (INSERM), HEMATIM - Hématopoïèse et immunologie - UR UPJV 4666 (HEMATIM), CHU Amiens-Picardie-Université de Picardie Jules Verne (UPJV)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Neuro-Immunologie Virale - Viral Neuro-immunology, This work was funded by the Vaincre les Maladies Lysosomales (VML) charity., We thank Professor Elizabeth Neufeld for providing the MPSIIIB mice, and Dr. Bénédicte Heron (APHP, Hôpital Trousseau, Paris, France) for collecting urine samples from patients. We are grateful to Camille Rottier (CHU Amiens Picardie, Amiens, France), Dr. Julie Bruyère (Institut des Neurosciences, Grenoble) and Nathalie Dessendier (Centre Français des Porphyries ‐ Laboratoire de Biochimie, Hôpital Louis Mourier, Colombes, France) for excellent technical assistance. We thank the staff of ICAP plateform, Dr. Paulo Marcelo and Mrs. Luciane Zabijak, for assistance with confocal microscopy experiments (ICAP plateform, Université de Picardie Jules Verne, Amiens, France)., Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Mécanismes physiopathologiques et conséquences des calcifications vasculaires - UR UPJV 7517 (MP3CV), and Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie
Subjects: 0301 basic medicine, Sanfilippo syndrome, MESH: Neurons, microglia, MESH: Mice, Knockout, [SDV.MHEP.UN]Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology, neuroinflammation, MESH: Neurodegenerative Diseases, Mucopolysaccharidosis III, chemistry.chemical_compound, iron, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Animals, MESH: Mucopolysaccharidosis III, Mice, Knockout, Neurons, MESH: Iron, Microglia, Brain, Neurodegenerative Diseases, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, Heparan sulfate, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, MESH: Microglia, medicine.anatomical_structure, Neurology, Cerebral cortex, medicine.medical_specialty, Mucopolysaccharidosis type III, Biology, MESH: Brain, 03 medical and health sciences, Cellular and Molecular Neuroscience, Hepcidin, Internal medicine, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], medicine, Animals, Neuroinflammation, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, medicine.disease, MESH: Astrocytes, 030104 developmental biology, Endocrinology, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, chemistry, Astrocytes, TLR4, biology.protein, hepcidin, 030217 neurology & neurosurgery
Abstract: International audience; Neuroinflammation and iron accumulation are hallmarks of a variety of adult neurodegenerative diseases. In Sanfilippo syndrome (mucopolysaccharidosis type III, MPSIII, a pediatric neurodegenerative disease that shares some features with adult neurodegenerative diseases), the progressive accumulation of heparan sulfate oligosaccharides (HSOs) induces microglia and astrocytes to produce pro-inflammatory cytokines leading to severe neuroinflammation. The objectives of the present study were (1) to measure the local iron concentration and to assess iron metabolism in the brain of a MPSIIIB murine model and (2) to identify the brain cells involved in this accumulation. We found that iron accumulation in MPSIIIB mice primarily affected the cerebral cortex where hepcidin levels were higher than in wild-type mice, and increased with aging. This increase was correlated with low expression of ferroportin 1 (FPN1), and thus brain iron retention. Moreover, we showed in vitro that HSOs are directly responsible for the production of hepcidin and the relative decrease in FPN1 expression when added to cultures of microglia and, to a lesser extent, to cultures of astrocytes. In contrast, no significant differences were observed in neurons. Hepcidin induction results from activation of the TLR4 pathway and STAT3 signaling, and leads to iron retention within microglia. Our results show that microglia have a key role in cerebral hepcidin overexpression and thus in the brain iron accumulation observed in the MPSIIIB model.
Published: 2018

24. Guanine nucleotide exchange factor RABGEF1 regulates keratinocyte-intrinsic signaling to maintain skin homeostasis

Author: Mindy Tsai, Nicolas Gaudenzio, Axel Roers, See-Ying Tam, Yuko Kawakami, Pierre Chambon, Philipp Starkl, Dimitri Pirottin, Stephen J. Galli, Sophie El Abbas, Jinah Kim, Nadine Antoine, Fabrice Bureau, Oliwia W Zurek, Laurent L. Reber, Toshiaki Kawakami, Thomas Marichal, Riccardo Sibilano, Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA-Research), Université de Liège, Department of Pathology [Stanford], Stanford Medicine, Stanford University-Stanford University, Sean N. Parker Center for Allergy and Asthma Research [Stanford], Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Technische Universität Dresden = Dresden University of Technology (TU Dresden), Cell and Developmental Biology [San Diego], School of Biological Sciences [Univ California San Diego] (UC San Diego), University of California [San Diego] (UC San Diego), University of California (UC)-University of California (UC)-University of California [San Diego] (UC San Diego), University of California (UC)-University of California (UC), Environmental Molecular Biology Laboratory (RIKEN), RIKEN - Institute of Physical and Chemical Research [Japon] (RIKEN), Walloon Excellence in Life sciences and BIOtechnology [Liège] (WELBIO), Stanford School of Medicine [Stanford], Department of Microbiology and Immunology [Stanford], and Pistre, Karine
Subjects: Keratinocytes, 0301 basic medicine, MESH: Guanine Nucleotide Exchange Factors / genetics, [SDV]Life Sciences [q-bio], MESH: Myeloid Differentiation Factor 88 / immunology, MESH: Dermatitis, Contact / immunology, Dermatitis, Contact, MESH: Mice, Knockout, Mice, 0302 clinical medicine, MESH: Humans Keratinocytes / immunology, Guanine Nucleotide Exchange Factors, MESH: Animals, MESH: NF-kappa B / genetics, Mice, Knockout, MESH: Dermatitis, Contact / pathology, integumentary system, MESH: Signal Transduction / immunology, MESH: Guanine Nucleotide Exchange Factors / immunology, Chemistry, NF-kappa B, MESH: Epidermis / pathology, General Medicine, Atopic dermatitis, Phenotype, Cell biology, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, MESH: NF-kappa B / immunology, 030220 oncology & carcinogenesis, MESH: Dermatitis, Contact / genetics, [SDV.IMM]Life Sciences [q-bio]/Immunology, Female, Guanine nucleotide exchange factor, MESH: Signal Transduction / genetics Signal Transduction / immunology, Signal transduction, medicine.symptom, Keratinocyte, Signal Transduction, Research Article, [SDV.IMM] Life Sciences [q-bio]/Immunology, Inflammation, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, 03 medical and health sciences, MESH: Keratinocytes / pathology, medicine, Animals, Humans, MESH: Receptors, Interleukin-1 Type I / immunology, MESH: Mice, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Allergic contact dermatitis, Receptors, Interleukin-1 Type I, medicine.disease, 030104 developmental biology, Myeloid Differentiation Factor 88, Rab, Epidermis, MESH: Receptors, Interleukin-1 Type I / genetics, MESH: Female, MESH: Myeloid Differentiation Factor 88 / genetics
Abstract: International audience; Epidermal keratinocytes form a structural and immune barrier that is essential for skin homeostasis. However, the mechanisms that regulate epidermal barrier function are incompletely understood. Here we have found that keratinocyte-specific deletion of the gene encoding RAB guanine nucleotide exchange factor 1 (RABGEF1, also known as RABEX-5) severely impairs epidermal barrier function in mice and induces an allergic cutaneous and systemic phenotype. RABGEF1-deficient keratinocytes exhibited aberrant activation of the intrinsic IL-1R/MYD88/NF-κB signaling pathway and MYD88-dependent abnormalities in expression of structural proteins that contribute to skin barrier function. Moreover, ablation of MYD88 signaling in RABGEF1-deficient keratinocytes or deletion of Il1r1 restored skin homeostasis and prevented development of skin inflammation. We further demonstrated that epidermal RABGEF1 expression is reduced in skin lesions of humans diagnosed with either atopic dermatitis or allergic contact dermatitis as well as in an inducible mouse model of allergic dermatitis. Our findings reveal a key role for RABGEF1 in dampening keratinocyte-intrinsic MYD88 signaling and sustaining epidermal barrier function in mice, and suggest that dysregulation of RABGEF1 expression may contribute to epidermal barrier dysfunction in allergic skin disorders in mice and humans. Thus, RABGEF1-mediated regulation of IL-1R/MYD88 signaling might represent a potential therapeutic target.
Published: 2016

25. Hormonal and spatial control of SUMOylation in the human and mouse adrenal cortex

Author: T. Dumontet, Antoine Martinez, Bruno Ragazzon, Anne-Marie Lefrançois-Martinez, Jean-Christophe Pointud, Nathanaëlle Montanier, Jérôme Bertherat, Igor Tauveron, Florence Roucher-Boulez, Isabelle Sahut-Barnola, Annabel Berthon, Damien Dufour, Pierre Val, Cyril Djari, M. Batisse-Lignier, Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), Interactions génétiques et cellulaires au cours de la différenciation, Université d'Auvergne - Clermont-Ferrand I (UdA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Génétique, Reproduction et Développement (GReD), Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Institut National de la Santé et de la Recherche Médicale (INSERM), Service d’endocrinologie diabétologie et maladies métaboliques, CHU Clermont-Ferrand, CHU Lyon, Unité de Nutrition Humaine (UNH), Institut National de la Recherche Agronomique (INRA)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), Génétique, Reproduction et Développement (GReD ), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), Service d’Endocrinologie Diabète et Maladies Métaboliques [CHU Clermont-Ferrand], Pôle RHEUNNIRS [CHU Clermont-Ferrand], CHU Gabriel Montpied [Clermont-Ferrand], CHU Clermont-Ferrand-CHU Clermont-Ferrand-CHU Gabriel Montpied [Clermont-Ferrand], CHU Clermont-Ferrand-CHU Clermont-Ferrand, Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), CHU Clermont-Ferrand, Centre de référence des maladies rares de la surrénale ( CHU Cochin [AP-HP]), Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Génétique, Reproduction et Développement - Clermont Auvergne (GReD), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne (UCA)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Clermont Université, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre National de la Recherche Scientifique (CNRS), ANR-16-IDEX-0001,CAP 20-25,CAP 20-25(2016), and Clermont Université-Université d'Auvergne - Clermont-Ferrand I (UdA)-Institut National de la Recherche Agronomique (INRA)
Subjects: 0301 basic medicine, MESH: Neoplasm Proteins, MESH: Signal Transduction, MESH: beta Catenin, SUMO protein, MESH: Adrenal Cortex Neoplasms, Biochemistry, MESH: Mice, Knockout, Dexamethasone, MESH: Zona Fasciculata, Mice, 0302 clinical medicine, Cortex (anatomy), PKA, MESH: Animals, Cycloheximide, MESH: Cycloheximide, Wnt Signaling Pathway, beta Catenin, ComputingMilieux_MISCELLANEOUS, Mice, Knockout, Adrenal cortex, MESH: Sumoylation, MESH: Wnt Signaling Pathway, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, PIAS3.β-catenin, Cell biology, Neoplasm Proteins, medicine.anatomical_structure, MESH: Dexamethasone, Dactinomycin, Female, Zona Glomerulosa, Signal transduction, Biotechnology, Endocrine gland, Signal Transduction, MESH: Colforsin, MESH: Cell Line, Tumor, MESH: Mice, Transgenic, MESH: Zona Glomerulosa, MESH: Delayed-Action Preparations, Repressor, Mice, Transgenic, [SDV.CAN]Life Sciences [q-bio]/Cancer, MESH: Cyclic AMP-Dependent Protein Kinases, Biology, 03 medical and health sciences, MESH: Carney Complex, Adrenocorticotropic Hormone, Cell Line, Tumor, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Genetics, medicine, Animals, Humans, mouse models, Carney Complex, Molecular Biology, Psychological repression, MESH: Mice, MESH: Adrenocorticotropic Hormone, MESH: Humans, Colforsin, Sumoylation, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, endocrine diseases, Cyclic AMP-Dependent Protein Kinases, Adrenal Cortex Neoplasms, MESH: Dactinomycin, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, 030104 developmental biology, SENP1/2, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Delayed-Action Preparations, MESH: Protein Processing, Post-Translational, Adrenal Cortex, MESH: Adrenal Cortex, Zona Fasciculata, Protein Processing, Post-Translational, MESH: Female, 030217 neurology & neurosurgery, Homeostasis
Abstract: SUMOylation is a highly conserved and dynamic post-translational mechanism primarily affecting nuclear programs for adapting organisms to stressful challenges. Alteration of SUMOylation cycles leads to severe developmental and homeostatic defects and malignancy, but signals coordinating SUMOylation are still unidentified. The adrenal cortex is a zonated endocrine gland that controls body homeostasis and stress response. Here, we show that in human and in mouse adrenals, SUMOylation follows a decreasing centripetal gradient that mirrors cortical differentiation flow and delimits highly and weakly SUMOylated steroidogenic compartments, overlapping glomerulosa, and fasciculata zones. Activation of PKA signaling by acute hormonal treatment, mouse genetic engineering, or in Carney complex results in repression of small ubiquitin-like modifier (SUMO) conjugation in the inner cortex by coordinating expression of SUMO pathway inducers and repressors. Conversely, genetic activation of canonical wingless-related integration site signaling maintains high SUMOylation potential in the outer neoplastic cortex. Thus, SUMOylation is tightly regulated by signaling pathways that orchestrate adrenal zonation and diseases.-Dumontet, T., Sahut-Barnola, I., Dufour, D., Lefrancois-Martinez, A.-M., Berthon, A., Montanier, N., Ragazzon, B., Djari, C., Pointud, J.-C., Roucher-Boulez, F., Batisse-Lignier, M., Tauveron, I., Bertherat, J., Val, P., Martinez, A. Hormonal and spatial control of SUMOylation in the human and mouse adrenal cortex.
Published: 2019

26. Postnatal Tshz3 deletion drives altered corticostriatal function and autism spectrum disorder-like behavior

Author: Michèle Carlier, Pierre L. Roubertoux, Christina Frahm, Laurent Fasano, Mehdi Metwaly, Dany Severac, Paolo Gubellini, Alistair N. Garratt, Bernard Jacq, Ahmed Fatmi, Pascal Salin, Dorian Chabbert, Xavier Caubit, Bianca Habermann, Lydia Kerkerian-Le Goff, Emeric Dubois, Institut de Biologie du Développement de Marseille (IBDM), Aix Marseille Université (AMU)-Collège de France (CdF (institution))-Centre National de la Recherche Scientifique (CNRS), Marseille medical genetics - Centre de génétique médicale de Marseille (MMG), Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU), Laboratoire de psychologie cognitive (LPC), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Theories and Approaches of Genomic Complexity (TAGC), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de génétique et physiologie du développement (LGPD), Université de la Méditerranée - Aix-Marseille 2-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Max Delbrück Center for Molecular Medicine [Berlin] (MDC), Helmholtz-Gemeinschaft = Helmholtz Association, Charité - UniversitätsMedizin = Charité - University Hospital [Berlin], Institut de Génomique Fonctionnelle - Montpellier GenomiX (IGF MGX), Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), ANR-17-CE16-0030,TSHZ3inASD,Rôle de TSHZ3 dans le développement et la fonction de systèmes neuronaux impliqués dans les troubles du spectre autistique(2017), ANR-10-INBS-0004,France-BioImaging,Développment d'une infrastructure française distribuée coordonnée(2010), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), ANR-11-IDEX-0001,Amidex,INITIATIVE D'EXCELLENCE AIX MARSEILLE UNIVERSITE(2011), ANR-17-EURE-0029,nEURo*AMU,Marseille NeuroSchool, une formation d'excellence(2017), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-BioCampus (BCM), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, 0301 basic medicine, MESH: Chromosomes, Human, Pair 19, [SDV]Life Sciences [q-bio], [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Haploinsufficiency, Striatum, MESH: Mice, Knockout, MESH: Synapses, Mice, 0302 clinical medicine, MESH: Behavior, Animal, MESH: Gene Expression Regulation, Developmental, MESH: Animals, Autism spectrum disorder, MESH: Heterozygote, Mice, Knockout, MESH: Autism Spectrum Disorder, Behavior, Animal, Gene Expression Regulation, Developmental, MESH: Transcription Factors, 3. Good health, TSHZ3, Synaptopathy, Knockout mouse, Cortex, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Glutamatergic synapse, MESH: Haploinsufficiency, Chromosome Deletion, Function and Dysfunction of the Nervous System, Heterozygote, MESH: Chromosome Deletion, Biology, behavioral disciplines and activities, 03 medical and health sciences, mental disorders, MESH: Homeodomain Proteins, medicine, Animals, Humans, MESH: Mice, Biological Psychiatry, Homeodomain Proteins, MESH: Humans, Sociability, medicine.disease, MESH: Male, Disease Models, Animal, 030104 developmental biology, Stereotypies, MESH: Gene Deletion, Synapses, Homeobox, MESH: Disease Models, Animal, Chromosomes, Human, Pair 19, Neuroscience, MESH: Female, Gene Deletion, 030217 neurology & neurosurgery, Transcription Factors
Abstract: International audience; BACKGROUND: Heterozygous deletion of the TSHZ3 gene, encoding for the teashirt zinc-finger homeobox family member 3 (TSHZ3) transcription factor that is highly expressed in cortical projection neurons (CPNs), has been linked to an autism spectrum disorder (ASD) syndrome. Similarly, mice with Tshz3 haploinsufficiency show ASD-like behavior, paralleled by molecular changes in CPNs and corticostriatal synaptic dysfunctions. Here, we aimed at gaining more insight into "when" and "where" TSHZ3 is required for the proper development of the brain, and its deficiency crucial for developing this ASD syndrome. METHODS: We generated and characterized a novel mouse model of conditional Tshz3 deletion, obtained by crossing Tshz3 flox/flox with CaMKIIalpha-Cre mice, in which Tshz3 is deleted in CPNs from postnatal day 2 to 3 onward. We characterized these mice by a multilevel approach combining genetics, cell biology, electrophysiology, behavioral testing, and bioinformatics. RESULTS: These conditional Tshz3 knockout mice exhibit altered cortical expression of more than 1000 genes, w50% of which have their human orthologue involved in ASD, in particular genes encoding for glutamatergic syn-apse components. Consistently, we detected electrophysiological and synaptic changes in CPNs and impaired corticostriatal transmission and plasticity. Furthermore, these mice showed strong ASD-like behavioral deficits. CONCLUSIONS: Our study reveals a crucial postnatal role of TSHZ3 in the development and functioning of the corticostriatal circuitry and provides evidence that dysfunction in these circuits might be determinant for ASD pathogenesis. Our conditional Tshz3 knockout mouse constitutes a novel ASD model, opening the possibility for an early postnatal therapeutic window for the syndrome linked to TSHZ3 haploinsufficiency.
Published: 2019

27. A critical role for miR-142 in alveolar epithelial lineage formation in mouse lung development

Author: Ana Ivonne Vazquez-Armendariz, Bernard Mari, Indrabahadur Singh, Nicolas Nottet, Rory E. Morty, Amit Shrestha, Cho-Ming Chao, Jochen Wilhelm, Jin San Zhang, Guillermo Barreto, Elie El Agha, Saverio Bellusci, Susanne Herold, Gianni Carraro, Chengshui Chen, Julio Cordero, Wenzhou Medical University [Wenzhou, China] (WMU), Justus-Liebig-Universität Gießen (JLU), Cedars-Sinai Medical Center, Institut de pharmacologie moléculaire et cellulaire (IPMC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), Université Côte d'Azur (UCA), Max Planck Institute for Heart and Lung Research (MPI-HLR), Max-Planck-Gesellschaft, Kazan Federal University (KFU), Department of Internal Medicine II [Giessen, Germany] (Cardio-Pulmonary Institute), University of Giessen Lung Center [Giessen, Germany], Institut Sophia Agrobiotech [Sophia Antipolis] (ISA), Institut National de la Recherche Agronomique (INRA)-Université Nice Sophia Antipolis (... - 2019) (UNS), Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA)-Centre National de la Recherche Scientifique (CNRS), Université Nice Sophia Antipolis (... - 2019) (UNS), Department of Pulmonary and Critical Care Medicine [Wenzhou, China], The First Affiliated Hospital of Wenzhou Medical University [Wenzhou, China], Institut Sophia Agrobiotech (ISA), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Recherche Agronomique (INRA), and COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS)
Subjects: MAPK/ERK pathway, Organogenesis, [SDV]Life Sciences [q-bio], Cell Maturation, MESH: Mice, Knockout, Mice, MESH: Respiratory Mucosa, MESH: Animals, Lung, Cells, Cultured, ComputingMilieux_MISCELLANEOUS, Mice, Knockout, EP300, 0303 health sciences, microRNA-142, 030302 biochemistry & molecular biology, respiratory system, Cell biology, medicine.anatomical_structure, cardiovascular system, Molecular Medicine, Stem cell, hormones, hormone substitutes, and hormone antagonists, circulatory and respiratory physiology, MESH: Cells, Cultured, Beta-catenin, Alveolar Epithelium, Respiratory Mucosa, Biology, MESH: Alveolar Epithelial Cells, 03 medical and health sciences, Cellular and Molecular Neuroscience, medicine, Animals, Cell Lineage, MESH: Lung, Progenitor cell, Molecular Biology, MESH: Mice, 030304 developmental biology, Pharmacology, Cell Biology, MESH: Cell Lineage, MicroRNAs, MESH: Organogenesis, Alveolar epithelium, Alveolar Epithelial Cells, biology.protein, Respiratory epithelium, MESH: MicroRNAs
Abstract: International audience; The respiratory epithelium arises from alveolar epithelial progenitors which differentiate into alveolar epithelial type 1 (AT1) and type 2 (AT2) cells. AT2 cells are stem cells in the lung critical for the repair process after injury. Mechanisms regulating AT1 and AT2 cell maturation are poorly defined. We report that the activation of the glucocorticoid pathway in an in vitro alveolar epithelial lineage differentiation assay led to increased AT2 marker Sftpc and decreased miR-142 expression. Using miR-142 KO mice, we demonstrate an increase in the AT2/AT1 cell number ratio. Overexpression of miR-142 in alveolar progenitor cells in vivo led to the opposite effect. Examination of the KO lungs at E18.5 revealed enhanced expression of miR-142 targets Apc, Ep300 and Kras associated with increased β-catenin and p-Erk signaling. Silencing of miR-142 expression in lung explants grown in vitro triggers enhanced Sftpc expression as well as increased AT2/AT1 cell number ratio. Pharmacological inhibition of Ep300-β-catenin but not Erk in vitro prevented the increase in Sftpc expression triggered by loss of miR-142. These results suggest that the glucocorticoid-miR-142-Ep300-β-catenin signaling axis controls pneumocyte maturation.
Published: 2019

28. Hepatocyte-specific loss of GPS2 in mice reduces non-alcoholic steatohepatitis via activation of PPARα

Author: Tarja Schröder, Matteo Pedrelli, Luc Van Gaal, Anthony N. Hollenberg, Arturo Mendoza, Philippe Lefebvre, Ning Liang, Saioa Goñi, Zhiqiang Huang, Bart Staels, Raoul Kuiper, Paolo Parini, Serena Barilla, Lise Lotte Vedin, Eckardt Treuter, Nicolas Venteclef, Sven Francque, Tomas Jakobsson, Anastasius Damdimopoulos, Marco Giudici, Rongrong Fan, Fawaz Alzaid, Osman Ahmed, Department of Biosciences and Nutrition [Karolinska Institutet], Karolinska Institutet [Stockholm], Department of Laboratory Medicine [Karolinska Institutet], Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Weill Medical College of Cornell University [New York], Department of Medicine Huddinge [Karolinska Institutet], Inflammation and Infection Theme [Karolinska University Hospital], Karolinska University Hospital [Stockholm], Récepteurs nucléaires, maladies cardiovasculaires et diabète - U 1011 (RNMCD), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Gastroenterology and Hepatology, University of Antwerp (UA), Laboratory of Experimental Medicine and Pediatrics, Department of Endocrinology, Diabetology and Metabolism, E.T. was supported by grants from the Center for Innovative Medicine (CIMED) at the Karolinska Institutet, the Swedish Research Council, the Swedish Cancer Society, the Swedish Diabetes Foundation, the Novo Nordisk Foundation, and the European Union FP7 HEALTH project HUMAN. R.F. was supported by the European Foundation for the Study of Diabetes (EFSD)/Lilly research fellowship, Junior Diabetes Wellness Sverige grant and KI research foundation grants. R.K. and T.S. were supported by KI/SL and grants from CIMED. N.L. received a doctoral education grant (KID) from Karolinska Institutet. Z.H. was supported by Chinese Government Scholarship (CSC). N.V. was supported by grants from the French National Agency of Research (CONRAD and PROVIDE), Region Ile de France (CORDDIM), Paris city (EMERGENCE), the French Foundation for Diabetes (SFD), and the European Union H2020 framework (ERC-EpiFAT 725790). The human liver transcriptome study performed by P.L., F.A., N.V., and B.S. was supported by EU [HEPADIP (Contract LSHM-CT-2005-018734) and RESOLVE (Contract FP7-305707)], the European Research Council (ERC Grant Immunobile, contract 694717), Fondation pour la Recherche Médicale (Equipe labellisée, DEQ20150331724) and Agence Nationale pour la Recherche (ANR-10-LBEX-46). B.S. is a recipient of an Advanced ERC Grant (694717), ANR-12-JSV1-0007,CONRAD,Le complexe répresseur SMRT-GPS2: senseur moléculaire de l'inflammation du tissu adipeux dans l'obésité humaine(2012), ANR-15-CE14-0029,PROVIDE,Compréhension des mécanismes inflammatoires protecteurs dans le diabète(2015), European Project: 694717,H2020-EU.1.1. - EXCELLENT SCIENCE - European Research Council (ERC) ,ImmunoBile(2016), École Pratique des Hautes Études (EPHE), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Derudas, Marie-Hélène, Jeunes Chercheuses et Jeunes Chercheurs - Le complexe répresseur SMRT-GPS2: senseur moléculaire de l'inflammation du tissu adipeux dans l'obésité humaine - - CONRAD2012 - ANR-12-JSV1-0007 - JC - VALID, Compréhension des mécanismes inflammatoires protecteurs dans le diabète - - PROVIDE2015 - ANR-15-CE14-0029 - AAPG2015 - VALID, and Bile acid, immune-metabolism, lipid and glucose homeostasis - ImmunoBile - - H2020-EU.1.1. - EXCELLENT SCIENCE - European Research Council (ERC) 2016-09-01 - 2021-08-31 - 694717 - VALID
Subjects: Male, 0301 basic medicine, Biopsy, [SDV]Life Sciences [q-bio], Datasets as Topic, General Physics and Astronomy, 02 engineering and technology, MESH: Mice, Knockout, Epigenesis, Genetic, MESH: Hepatocytes, Mice, MESH: Biopsy, Non-alcoholic Fatty Liver Disease, MESH: Animals, MESH: Datasets as Topic, MESH: Epigenesis, Genetic, lcsh:Science, MESH: PPAR alpha, MESH: Lipid Metabolism, Mice, Knockout, Multidisciplinary, Chemistry, Fatty liver, Intracellular Signaling Peptides and Proteins, 021001 nanoscience & nanotechnology, 3. Good health, GPS2, [SDV] Life Sciences [q-bio], Liver, Cistrome, MESH: HEK293 Cells, MESH: Fibrosis, Disease Progression, MESH: Disease Progression, Peroxisome proliferator-activated receptor alpha, 0210 nano-technology, Engineering sciences. Technology, Science, Diet, High-Fat, Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, MESH: Mice, Inbred C57BL, MESH: Intracellular Signaling Peptides and Proteins, medicine, Animals, Humans, PPAR alpha, MESH: Mice, MESH: Humans, MESH: Non-alcoholic Fatty Liver Disease, General Chemistry, Lipid Metabolism, medicine.disease, HDAC3, Fibrosis, MESH: Male, Mice, Inbred C57BL, Disease Models, Animal, MESH: Diet, High-Fat, HEK293 Cells, 030104 developmental biology, Nuclear receptor, Hepatocytes, Cancer research, lcsh:Q, Steatosis, Steatohepatitis, MESH: Disease Models, Animal, MESH: Liver
Abstract: Obesity triggers the development of non-alcoholic fatty liver disease (NAFLD), which involves alterations of regulatory transcription networks and epigenomes in hepatocytes. Here we demonstrate that G protein pathway suppressor 2 (GPS2), a subunit of the nuclear receptor corepressor (NCOR) and histone deacetylase 3 (HDAC3) complex, has a central role in these alterations and accelerates the progression of NAFLD towards non-alcoholic steatohepatitis (NASH). Hepatocyte-specific Gps2 knockout in mice alleviates the development of diet-induced steatosis and fibrosis and causes activation of lipid catabolic genes. Integrative cistrome, epigenome and transcriptome analysis identifies the lipid-sensing peroxisome proliferator-activated receptor α (PPARα, NR1C1) as a direct GPS2 target. Liver gene expression data from human patients reveal that Gps2 expression positively correlates with a NASH/fibrosis gene signature. Collectively, our data suggest that the GPS2-PPARα partnership in hepatocytes coordinates the progression of NAFLD in mice and in humans and thus might be of therapeutic interest., Dysregulation of PPARα dependent fatty acid oxidation promotes hepatic steatosis. Here the authors show that GPS2 inhibits PPARα activity and that ablation of GPS2 ameliorates hepatic steatosis in mice.
Published: 2019

29. IFN-γ is a therapeutic target in paraneoplastic cerebellar degeneration

Author: Roland S. Liblau, Béatrice Pignolet, Emilie Mauré, Monika C. Brunner-Weinzierl, L. M. Yshii, Jan Bauer, Mandy Pierau, Oliver Hartley, Centre de Physiopathologie Toulouse Purpan (CPTP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Otto-von-Guericke-Universität Magdeburg = Otto-von-Guericke University [Magdeburg] (OVGU), Université de Genève = University of Geneva (UNIGE), Medizinische Universität Wien = Medical University of Vienna, ANR-13-BSV3-0003,T cell Mig,Migration des lymp^hocytes T CD8 T dans le système nerveux central(2013), Pistre, Karine, and Blanc 2013 - Migration des lymp^hocytes T CD8 T dans le système nerveux central - - T cell Mig2013 - ANR-13-BSV3-0003 - Blanc 2013 - VALID
Subjects: 0301 basic medicine, Cerebellum, Tumor/transplantation, Integrin alpha4, T-Lymphocytes, Autoimmune diseases, MESH: Integrin alpha4 / metabolism, [SDV]Life Sciences [q-bio], Autoimmunity, ddc:616.07, medicine.disease_cause, Autoantigens, MESH: Mice, Knockout, Mice, Purkinje Cells, 0302 clinical medicine, Breast cancer, MESH: Mammary Neoplasms, Experimental / immunology, MESH: Interferon-gamma / metabolism, Cancer, Mice, Knockout, biology, Neoplasm/immunology, Experimental/complications/immunology, Purkinje Cells/immunology/metabolism/pathology, General Medicine, Paraneoplastic cerebellar degeneration, Integrin alpha4/antagonists & inhibitors/immunology/metabolism, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, MESH: Paraneoplastic Cerebellar Degeneration / drug therapy, 030220 oncology & carcinogenesis, MESH: Autoantigens / immunology, [SDV.IMM]Life Sciences [q-bio]/Immunology, Female, Antibody, Autoantibodies/immunology, Research Article, MESH: T-Lymphocytes / metabolism, MESH: Interferon-gamma / immunology, MESH: Interferon-gamma / antagonists & inhibitors, MESH: Paraneoplastic Cerebellar Degeneration / pathology, MESH: T-Lymphocytes / immunology, [SDV.IMM] Life Sciences [q-bio]/Immunology, Knockout, Integrin, Immunology, MESH: Purkinje Cells / metabolism, [SDV.CAN]Life Sciences [q-bio]/Cancer, Interferon-gamma/antagonists & inhibitors/immunology/metabolism, Paraneoplastic Cerebellar Degeneration, Cell Line, MESH: Integrin alpha4 / immunology, 03 medical and health sciences, Interferon-gamma, T-Lymphocytes/drug effects/immunology/metabolism, [SDV.CAN] Life Sciences [q-bio]/Cancer, Autoantigens/immunology, Immunity, Antigens, Neoplasm, Cell Line, Tumor, medicine, Animals, MESH: Cell Line, Tumor / transplantation, Antigens, MESH: Mice, Autoantibodies, business.industry, Mammary Neoplasms, Autoantibody, MESH: Purkinje Cells / pathology, Mammary Neoplasms, Experimental, MESH: Integrin alpha4 / antagonists & inhibitors, medicine.disease, Immune checkpoint, 030104 developmental biology, MESH: Paraneoplastic Cerebellar Degeneration / immunology, Cancer research, biology.protein, MESH: Mammary Neoplasms, Experimental / complications, MESH: Purkinje Cells / immunology, MESH: T-Lymphocytes / drug effects, business, Paraneoplastic Cerebellar Degeneration/drug therapy/immunology/pathology, MESH: Female
Abstract: International audience; Paraneoplastic neurological disorders result from an autoimmune response against neural self-antigens that are ectopically expressed in neoplastic cells. In paraneoplastic disorders associated to autoantibodies against intracellular proteins, such as paraneoplastic cerebellar degeneration (PCD), current data point to a major role of cell-mediated immunity. In an animal model, in which a neo-self-antigen was expressed in both Purkinje neurons and implanted breast tumor cells, immune checkpoint blockade led to complete tumor control at the expense of cerebellum infiltration by T cells and Purkinje neuron loss, thereby mimicking PCD. Here, we identify 2 potential therapeutic targets expressed by cerebellum-infiltrating T cells in this model, namely α4 integrin and IFN-γ. Mice with PCD were treated with anti-α4 integrin antibodies or neutralizing anti-IFN-γ antibodies at the onset of neurological signs. Although blocking α4 integrin had little or no impact on disease development, treatment using the anti-IFN-γ antibody led to almost complete protection from PCD. These findings strongly suggest that the production of IFN-γ by cerebellum-invading T cells plays a major role in Purkinje neuron death. Our successful preclinical use of neutralizing anti-IFN-γ antibody for the treatment of PCD offers a potentially new therapeutic opportunity for cancer patients at the onset of paraneoplastic neurological disorders.
Published: 2019

30. Endothelial α1AMPK modulates angiotensin II-mediated vascular inflammation and dysfunction

Author: Andreas Daiber, Marc Foretz, Matthias Oelze, Sabine Kossmann, Philip Wenzel, Thomas Münzel, Jeremy Lagrange, Katie Frenis, John F. Keaney, Eberhard Schulz, Thi Lan P. Tran, Thomas Jansen, Benoit Viollet, Swenja Kröller-Schön, Miroslawa Kvandová, Sanela Kalinovic, Johannes Gutenberg - Universität Mainz (JGU), University of Massachusetts Medical School [Worcester] (UMASS), University of Massachusetts System (UMASS), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Paris 5 (UPD5), and Johannes Gutenberg - University of Mainz (JGU)
Subjects: Male, 0301 basic medicine, MESH: Inflammation, Physiology, AMP-Activated Protein Kinases, 030204 cardiovascular system & hematology, medicine.disease_cause, MESH: Mice, Knockout, Antioxidants, Mice, 0302 clinical medicine, MESH: Animals, Endothelial dysfunction, MESH: AMP-Activated Protein Kinases, Vascular inflammation, Mice, Knockout, NADPH oxidase, MESH: Oxidative Stress, biology, MESH: Heme Oxygenase-1, Chemistry, Angiotensin II, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, α1AMPK, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Integrin alpha M, Knockout mouse, MESH: Angiotensin II, MESH: Membrane Proteins, MESH: Endothelium, Vascular, Cardiology and Cardiovascular Medicine, medicine.medical_specialty, 03 medical and health sciences, Downregulation and upregulation, MESH: Mice, Inbred C57BL, Physiology (medical), Internal medicine, medicine, Animals, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], MESH: Mice, Inflammation, Macrophages, MESH: Antioxidants, Membrane Proteins, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, medicine.disease, MESH: Male, Mice, Inbred C57BL, Heme oxygenase, Oxidative Stress, 030104 developmental biology, Endocrinology, biology.protein, Endothelium, Vascular, Reactive oxygen species, Heme Oxygenase-1, Oxidative stress
Abstract: International audience; Mice with a global deletion of α1AMPK are characterized by endothelial dysfunction and NADPH oxidase subunit 2 (NOX-2)-mediated vascular oxidative stress. However, the underlying mechanisms are incompletely understood and may involve endothelial NOX-2 upregulation or facilitated vascular infiltration of phagocytic cells. Therefore, the current study was designed to investigate the vascular effects of chronic angiotensin II (AngII) infusion in mice with an endothelial-specific α1AMPK deletion. A mouse strain with endothelial-specific α1AMPK deletion was generated by breeding α1AMPKflox/flox mice with TekCre+ or Cadh5Cre+ mice. Chronic AngII infusion (0.5 mg/kg/day for 7day) caused mild endothelial dysfunction in wild-type mice that was significantly aggravated in endothelial α1AMPK knockout mice. Aortic NOX-2 and CD68 expression were increased, indicating that infiltrating leukocytes may significantly contribute to enhanced vascular oxidative stress. Flow cytometry revealed a higher abundance of aortic CD90.2+ T-cells, CD11b+F4/80+ macrophages and Ly6G-Ly6C+ monocytes. Vascular mRNA expression of monocyte chemoattractant protein 1, CCL5 and vascular cell adhesion molecule 1 was enhanced in AngII-infused mice lacking endothelial α1AMPK, facilitating the recruitment of inflammatory cells to the vessel wall. In addition, AngII-induced upregulation of cytoprotective heme oxygenase 1 (HO-1) was blunted in mice with endothelial α1AMPK deletion, compatible with an impaired antioxidant defense in these animals. In summary, endothelial expressed α1AMPK limits the recruitment of inflammatory cells to the vessel wall and maintains HO-1 mediated antioxidant defense. Both mechanisms reduce vascular oxidative damage and preserve endothelial function during chronic AngII treatment.
Published: 2019

31. Impaired telomere integrity and rRNA biogenesis in PARN-deficient patients and knock-out models

Author: Benyelles, Maname, Episkopou, Harikleia, O'Donohue, Marie-Françoise, Kermasson, Laetitia, Frange, Pierre, Poulain, Florian, Burcu Belen, Fatma, Polat, Meltem, Bole-Feysot, Christine, Langa-Vives, Francina, Gleizes, Pierre-Emmanuel, de Villartay, Jean-Pierre, Callebaut, Isabelle, Decottignies, Anabelle, Revy, Patrick, Laboratoire de Biologie Moléculaire de la Cellule (LBMC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Université catholique de Louvain, Institut des Sciences de la Vie, Imagine - Institut des maladies génétiques (IMAGINE - U1163), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), CHU Necker - Enfants Malades [AP-HP], Institut de minéralogie, de physique des matériaux et de cosmochimie (IMPMC), Muséum national d'Histoire naturelle (MNHN)-Institut de recherche pour le développement [IRD] : UR206-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Genetic and Epigenetic Alterations of Genomes (GEAG), Université Catholique de Louvain (UCL)-de Duve Institute, Developpement Normal et Pathologique du Système Immunitaire, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Genome dynamics in the immune system (Equipe Inserm U1163), Imagine - Institut des maladies génétiques (IHU) (Imagine - U1163), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Université Catholique de Louvain = Catholic University of Louvain (UCL), Laboratoire de biologie moléculaire eucaryote (LBME), Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Infection à VIH, réservoirs, diversité génétique et résistance aux antirétroviraux (ARV) (EA 7327), Université Paris Descartes - Paris 5 (UPD5), Başkent University Hospital [Adana, Turkey], Pamukkale University, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], This work has been supported by institutional grants from INSERM, Ligue Nationale contre le Cancer (Equipe Labellisée La Ligue), Institut National du Cancer INCa, GIS‐Institut des maladies rares, and FNRS (Fonds National de la Recherche Scientifique, Belgium). P.E.G. and M.F.O. are supported by Agence Nationale de la Recherche (ANR 2015 AAP générique CE12‐0001‐DBA Multigenes), and the EuroDBA project is funded by the ERA‐NET program E‐RAR3 (ANR‐15‐RAR3‐0007‐04). P.R. and M.F.O. are scientists from Centre National de la Recherche Scientifique (CNRS). H.E. and F.P. were supported by grants from the Télévie/FNRS and FRIA/FNRS. A.D. is a scientist from the FNRS., ANR-15-CE12-0001,DBA-MULTIGENES,DBA-MULTIGENES: identification et caractérisation de gènes candidats pour la découverte de nouveaux mécanismes physiopathologiques à l'origine de l'anémie de Blackfan-Diamond.(2015), ANR-15-RAR3-0007,EuroDBA,The European Diamond-Blackfan Anemia Consortium(2015), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Institut Pasteur [Paris] (IP), O'Donohue, Marie-Françoise, DBA-MULTIGENES: identification et caractérisation de gènes candidats pour la découverte de nouveaux mécanismes physiopathologiques à l'origine de l'anémie de Blackfan-Diamond. - - DBA-MULTIGENES2015 - ANR-15-CE12-0001 - AAPG2015 - VALID, The European Diamond-Blackfan Anemia Consortium - - EuroDBA2015 - ANR-15-RAR3-0007 - E-Rare-3 - VALID, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre de Biologie Intégrative (CBI), and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, p53, Medicine (General), [SDV]Life Sciences [q-bio], Høyeraal–Hreidarsson syndrome, QH426-470, MESH: Mice, Knockout, Chromatin, Epigenetics, Genomics & Functional Genomics, Shelterin Complex, Mice, MESH: Animals, rRNA, ComputingMilieux_MISCELLANEOUS, Mice, Knockout, Fetal Growth Retardation, Articles, Telomere, Child, Preschool, MESH: Exoribonucleases, Microcephaly, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Female, shelterin, PARN, MESH: Fetal Growth Retardation, Telomere-Binding Proteins, Høyeraal-Hreidarsson syndrome, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH: Microcephaly, Dyskeratosis Congenita, Article, MESH: Telomere Homeostasis, MESH: Intellectual Disability, R5-920, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Intellectual Disability, Genetics, Animals, Humans, MESH: Mice, MESH: Humans, MESH: Child, Preschool, Telomere Homeostasis, MESH: Male, Disease Models, Animal, MESH: Dyskeratosis Congenita, RNA, Ribosomal, Exoribonucleases, MESH: RNA, Ribosomal, Genetics, Gene Therapy & Genetic Disease, MESH: Disease Models, Animal, MESH: Telomere, MESH: Female
Abstract: International audience; PARN, poly(A)-specific ribonuclease, regulates the turnover of mRNAs and the maturation and stabilization of the hTR RNA component of telomerase. Biallelic PARN mutations were associated with Høyeraal-Hreidarsson (HH) syndrome, a rare telomere biology disorder that, because of its severity, is likely not exclusively due to hTR down-regulation. Whether PARN deficiency was affecting the expression of telomere-related genes was still unclear. Using cells from two unrelated HH individuals carrying novel PARN mutations and a human PARN knock-out (KO) cell line with inducible PARN complementation, we found that PARN deficiency affects both telomere length and stability and down-regulates the expression of TRF1, TRF2, TPP1, RAP1, and POT1 shelterin transcripts. Down-regulation of dyskerin-encoding DKC1 mRNA was also observed and found to result from p53 activation in PARN-deficient cells. We further showed that PARN deficiency compromises ribosomal RNA biogenesis in patients' fibroblasts and cells from heterozygous Parn KO mice. Homozygous Parn KO however resulted in early embryonic lethality that was not overcome by p53 KO. Our results refine our knowledge on the pleiotropic cellular consequences of PARN deficiency.
Published: 2018

32. High N-glycan multiplicity is critical for neuronal adhesion and sensitizes the developing cerebellum to N-glycosylation defect

Author: Medina-Cano, Daniel, Ucuncu, Ekin, Nguyen, Lam Son, Nicouleau, Michael, Lipecka, Joanna, Bizot, Jean-Charles, Thiel, Christian, Foulquier, François, Lefort, Nathalie, Faivre-Sarrailh, Catherine, Colleaux, Laurence, Guerrera, Ida Chiara, Cantagrel, Vincent, Imagine - Institut des maladies génétiques (IMAGINE - U1163), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de chimie et procédés pour l'énergie, l'environnement et la santé (ICPEES), Université de Strasbourg (UNISTRA)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), JRC Institute for Energy and Transport (IET), European Commission - Joint Research Centre [Petten], Institut de Neurobiologie de la Méditerranée [Aix-Marseille Université] (INMED - INSERM U1249), Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU), Structure Fédérative de Recherche Necker (SFR Necker - UMS 3633 / US24), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et nanosciences d'Alsace, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), PRA, Fanny, Mécanismes développementaux des anomalies structurelles cérébelleuses - - SCD-Mec2016 - ANR-16-CE12-0005 - AAPG2016 - VALID, ERA-NET rare disease research implementing IRDiRC objectives - E-Rare-3 - - H20202014-12-01 - 2019-11-30 - 643578 - VALID, Institut de psychiatrie et neurosciences (U894 / UMS 1266), Key-Obs, ANR-16-CE12-0005,SCD-Mec,Mécanismes développementaux des anomalies structurelles cérébelleuses(2016), European Project: 643578,H2020,H2020-HCO-2014,E-Rare-3(2014), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Lille, Centre de recherche en neurobiologie - neurophysiologie de Marseille (CRN2M), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Sorbonne Paris Cité (USPC), and Centre de Psychiatrie et Neurosciences (U894)
Subjects: Glycosylation, MESH: Immunoglobulins, [SDV]Life Sciences [q-bio], MESH: Neurons, N-glycosylation, MESH: Unfolded Protein Response, MESH: Mice, Knockout, neuroscience, Purkinje Cells, MESH: Axon Guidance, Neural Pathways, cell biology, MESH: Animals, Biology (General), Mice, Knockout, Neurons, neuronal migration, MESH: Proteomics, Srd5a3, Cell Differentiation, MESH: Glycosylation, MESH: Motor Activity, Axon Guidance, MESH: Reproducibility of Results, MESH: Cell Adhesion Molecules, Medicine, lipids (amino acids, peptides, and proteins), [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH: Membrane Proteins, Research Article, MESH: Cell Differentiation, MESH: Mutation, animal structures, cerebellum, QH301-705.5, Science, Induced Pluripotent Stem Cells, Immunoglobulins, MESH: Cytoplasmic Granules, Motor Activity, MESH: Induced Pluripotent Stem Cells, Cytoplasmic Granules, MESH: 3-Oxo-5-alpha-Steroid 4-Dehydrogenase, MESH: Cell Adhesion, MESH: Purkinje Cells, proteomics, 3-Oxo-5-alpha-Steroid 4-Dehydrogenase, Polysaccharides, Animals, human, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], mouse, MESH: Neural Pathways, Cell Membrane, Membrane Proteins, Reproducibility of Results, cell adhesion, MESH: Cerebellum, carbohydrates (lipids), MESH: Polysaccharides, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, MESH: Gene Deletion, Protein N-glycosylation, Mutation, Unfolded Protein Response, Cell Adhesion Molecules, Gene Deletion, MESH: Cell Membrane, congenital disorders of glycosylation
Abstract: International audience; Proper brain development relies highly on protein N-glycosylation to sustain neuronal migration, axon guidance and synaptic physiology. Impairing the N-glycosylation pathway at early steps produces broad neurological symptoms identified in congenital disorders of glycosylation. However, little is known about the molecular mechanisms underlying these defects. We generated a cerebellum specific knockout mouse for Srd5a3, a gene involved in the initiation of N-glycosylation. In addition to motor coordination defects and abnormal granule cell development, Srd5a3 deletion causes mild N-glycosylation impairment without significantly altering ER homeostasis. Using proteomic approaches, we identified that Srd5a3 loss affects a subset of glycoproteins with high N-glycans multiplicity per protein and decreased protein abundance or N-glycosylation level. As IgSF-CAM adhesion proteins are critical for neuron adhesion and highly N-glycosylated, we observed impaired IgSF-CAM-mediated neurite outgrowth and axon guidance in Srd5a3 mutant cerebellum. Our results link high N-glycan multiplicity to fine-tuned neural cell adhesion during mammalian brain development.
Published: 2018

33. Increased expression of ATP12A proton pump in cystic fibrosis airways

Author: Davide Tosi, Arianna Venturini, Luis J. V. Galietta, Alessandro Palleschi, Paolo Scudieri, Isabelle Sermet-Gaudelus, Gilles Crambert, Ilaria Musante, Pablo Martín-Vasallo, P. Morelli, Emanuela Caci, Gabrielle Planelles, Christine Walter, Centre de recherche Croissance et signalisation (UMR_S 845), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Espace et action, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche des Cordeliers (CRC), Université Pierre et Marie Curie - Paris 6 (UPMC)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratorio di Genetica Molecolare, Istituto G. Gaslini, Université Paris Diderot - Paris 7 (UPD7)-École pratique des hautes études (EPHE)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Scudieri, Paolo, Musante, Ilaria, Caci, Emanuela, Venturini, Arianna, Morelli, Patrizia, Walter, Christine, Tosi, Davide, Palleschi, Alessandro, Martin-Vasallo, Pablo, Sermet-Gaudelus, Isabelle, Planelles, Gabrielle, Crambert, Gille, and Galietta, Luis Jv
Subjects: 0301 basic medicine, MESH: Hydrogen-Ion Concentration, Cystic Fibrosis, Pulmonology, medicine.medical_treatment, [SDV]Life Sciences [q-bio], Genetic disease, Cystic fibrosis, MESH: Mice, Knockout, H(+)-K(+)-Exchanging ATPase, Mice, MESH: Ouabain, MESH: H(+)-K(+)-Exchanging ATPase, MESH: Animals, Ouabain, Cells, Cultured, MESH: Sodium-Potassium-Exchanging ATPase, Submucosal glands, Mice, Knockout, Cultured, Chemistry, MESH: Bronchi, Epithelial transport of ions and water, Genetic diseases, Animals, Bronchi, Cell Membrane, Colon, Goblet Cells, Humans, Hydrogen-Ion Concentration, Interleukin-4, Permeability, Potassium, Primary Cell Culture, Proton Pump Inhibitors, Sodium-Potassium-Exchanging ATPase, General Medicine, 3. Good health, Cytokine, MESH: Permeability, medicine.symptom, Research Article, MESH: Cells, Cultured, MESH: Proton Pump Inhibitors, MESH: Cystic Fibrosis, Cells, Knockout, Inflammation, MESH: Primary Cell Culture, 03 medical and health sciences, Downregulation and upregulation, medicine, Secretion, MESH: Colon, MESH: Mice, MESH: Humans, MESH: Interleukin-4, medicine.disease, Molecular biology, 030104 developmental biology, Cell culture, MESH: Potassium, Respiratory epithelium, MESH: Goblet Cells, MESH: Cell Membrane
Abstract: International audience; Proton secretion mediated by ATP12A protein on the surface of the airway epithelium may contribute to cystic fibrosis (CF) lung disease by favoring bacterial infection and airway obstruction. We studied ATP12A in fresh bronchial samples and in cultured epithelial cells. In vivo, ATP12A expression was found almost exclusively at the apical side of nonciliated cells of airway epithelium and in submucosal glands, with much higher expression in CF samples. This could be due to bacterial infection and inflammation, since treating cultured cells with bacterial supernatants or with IL-4 (a cytokine that induces goblet cell hyperplasia) increased the expression of ATP12A in nonciliated cells. This observation was associated with upregulation and translocation of ATP1B1 protein from the basal to apical epithelial side, where it colocalizes with ATP12A. ATP12A function was evaluated by measuring the pH of the apical fluid in cultured epithelia. Under resting conditions, CF epithelia showed more acidic values. This abnormality was minimized by inhibiting ATP12A with ouabain. Following treatment with IL-4, ATP12A function was markedly increased, as indicated by strong acidification occurring under bicarbonate-free conditions. Our study reveals potentially novel aspects of ATP12A and remarks its importance as a possible therapeutic target in CF and other respiratory diseases.
Published: 2018

34. MAVS deficiency induces gut dysbiotic microbiota conferring a proallergic phenotype

Author: Elsa Kress, Mathias Heikenwalder, Gerard Lina, David Bauché, Saskia Lippens, Claire Macari, Marie-Cécile Michallet, Mathias Chamaillard, Sophia Djebali, Emilie Plantamura, Lyvia Moudombi, Jacqueline Marvel, Christophe Caux, Julien C. Marie, Annabelle Cesaro, Lilia Boucinha, Amiran Dzutsev, Jean-Philippe Rasigade, Ulrike Rothermel, Giorgio Trinchieri, Azzak Bentaher-Belaaouaj, Oana Dumitrescu, Morgan Grau, Michelina Plateroti, Clovis Bondu, Immunité et lymphocytes cytotoxiques – Immunity and cytotoxic lymphocytes, Centre International de Recherche en Infectiologie (CIRI), École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet - Saint-Étienne (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Cancer Inflammation Program [Frederick], Center for Cancer Research, Leidos Biomedical Research, Inc [Frederick, MD, USA], Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), German Cancer Research Center - Deutsches Krebsforschungszentrum [Heidelberg] (DKFZ), Hospices Civils de Lyon (HCL), Pathogénie des Staphylocoques – Staphylococcal Pathogenesis (StaPath), VIB-UGent Center for Inflammation Research [Gand, Belgique] (IRC), VIB [Belgium], We acknowledge the contributions of the Structure Fédérative de Recherche BioSciences Gerland-Lyon Sud (UMS3444/US8-Ecole Normale Superieure de Lyon, Universite Claude Bernard de Lyon CNRS, INSERM) facilities, especially AniRA ImmOS. This work was supported by Institut National de la Santé et de la Recherche Médicale, Centre National de la Recherche Scientifique, Université de Lyon I, Hospices Civils de Lyon, Association 'ARCHE,' and Agence Nationale de la Recherche Grant ANR-11-RPIB-0019-03 (to J.M.), a grant from the Département du Rhône-Fonds Européen de Développement Régional (PLATINE) (J.M.), and European Commission Grant LSHG-CT-2006-037188, The European Mouse Disease Clinic (to J.M.)., We thank J. Tschopp’s laboratory for Cardif (MAVS-deficient) mice, Martine Tomkowiak, Julien Mafille, and Barbara Gilbert for expert technical assistance, Anca Hennino for confocal images, Christophe Arpin and Mohamad Sobh for statistical data analysis, Bertrand Dubois, Marc Vocanson, and Jean-François Nicolas for helpful discussions on the DTH model, Thierry Walzer, Laurent Genestier, Bertrand Dubois, and Gérard Eberl for critical reading of the manuscript, Bariza Blanquier for help with qPCR analysis (genetic analysis), Thibault Andrieu and Sébastien Dussurgey (AniRA-Cytometry), Olga Azocar and Christophe Chamot from the Plateau Technique Imagerie/Microcopie, and the staff of AniRA-PBES, especially Jean-Louis Thoumas and Céline Angleraux., Centre International de Recherche en Infectiologie - UMR (CIRI), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), and MICHALLET, MARIE-CECILE
Subjects: 0301 basic medicine, MESH: Signal Transduction, RIG-like receptors, MESH: Intestines/immunology, MESH: Skin Diseases, Bacterial/pathology, Gut flora, MESH: Skin Diseases, Bacterial/metabolism, MESH: Mice, Knockout, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Mice, 0302 clinical medicine, MESH: Animals, MESH: Hypersensitivity/metabolism, [SDV.IMM.ALL]Life Sciences [q-bio]/Immunology/Allergology, Sensitization, Mice, Knockout, Multidisciplinary, integumentary system, dysbiosis, Biological Sciences, MAVS, Phenotype, 3. Good health, Intestines, medicine.anatomical_structure, MESH: Intestines/pathology, 030220 oncology & carcinogenesis, [SDV.IMM]Life Sciences [q-bio]/Immunology, Female, [SDV.IMM.ALL] Life Sciences [q-bio]/Immunology/Allergology, Signal Transduction, MESH: Intestines/microbiology, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH: Dysbiosis/complications, Biology, MESH: Gastrointestinal Microbiome/immunology, MESH: Phenotype, digestive system, 03 medical and health sciences, MESH: Skin Diseases, Bacterial/etiology, MESH: Hypersensitivity/etiology, Immunity, MESH: Mice, Inbred C57BL, MESH: Homeodomain Proteins/metabolism, medicine, Hypersensitivity, allergic skin pathologies, MESH: Hypersensitivity/pathology, Animals, Colitis, Allergic contact dermatitis, [SDV.IMM.II] Life Sciences [q-bio]/Immunology/Innate immunity, MESH: Mice, Adaptor Proteins, Signal Transducing, Homeodomain Proteins, MESH: Adaptor Proteins, Signal Transducing/physiology, Intestinal permeability, Skin Diseases, Bacterial, medicine.disease, biology.organism_classification, Gastrointestinal Microbiome, Mice, Inbred C57BL, Disease Models, Animal, 030104 developmental biology, Immunology, MESH: Homeodomain Proteins/genetics, MESH: Disease Models, Animal, Dysbiosis, MESH: Female
Abstract: International audience; Prominent changes in the gut microbiota (referred to as "dysbiosis") play a key role in the development of allergic disorders, but the underlying mechanisms remain unknown. Study of the delayed-type hypersensitivity (DTH) response in mice contributed to our knowledge of the pathophysiology of human allergic contact dermatitis. Here we report a negative regulatory role of the RIG-I-like receptor adaptor mitochondrial antiviral signaling (MAVS) on DTH by modulating gut bacterial ecology. Cohousing and fecal transplantation experiments revealed that the dysbiotic microbiota of Mavs -/- mice conferred a proallergic phenotype that is communicable to wild-type mice. DTH sensitization coincided with increased intestinal permeability and bacterial translocation within lymphoid organs that enhanced DTH severity. Collectively, we unveiled an unexpected impact of RIG-I-like signaling on the gut microbiota with consequences on allergic skin disease outcome. Primarily, these data indicate that manipulating the gut microbiota may help in the development of therapeutic strategies for the treatment of human allergic skin pathologies.
Published: 2018

35. Aspergillus fumigatus conidial metalloprotease Mep1p cleaves host complement proteins

Author: Michel Monod, Oumaïma Ibrahim-Granet, Rajashri Shende, Rémi Beau, Jean-Paul Latgé, Karl-Heinz Gührs, Taruna Madan, Jayanta K. Pal, Arvind Sahu, Vishukumar Aimanianda, Sarah Sze Wah Wong, Srikanth Rapole, Savitribai Phule Pune University [Pune, india], Aspergillus, Institut Pasteur [Paris], Cytokines et Inflammation, Centre Hospitalier Universitaire Vaudois [Lausanne] (CHUV), Leibniz Institute on Aging - Fritz Lipmann Institute (FLI), Leibniz Association, National Institute for Research in Reproductive Health [Mumbai, India] (ICMR), This work was supported in part by a bilateral COMASPIN grant from the Department of Science and Technology (DST) India and l'Agence Nationale de la Recherché (ANR) France, and Institut Pasteur [Paris] (IP)
Subjects: 0301 basic medicine, metalloprotease, [SDV]Life Sciences [q-bio], Host Defense Mechanism, Biochemistry, MESH: Mice, Knockout, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Aspergillus fumigatus, MESH: Lectins, MESH: Collagen, complement, MESH: Animals, MESH: Immune Evasion, MESH: Phagocytosis, biology, Chemistry, phagocytosis, MESH: Complement C3, MESH: Complement C4, MESH: Gene Expression Regulation, Antibody opsonization, Aspergillus, MESH: Complement C5, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Immunity, Innate, MESH: Fungal Proteins, MESH: Aspergillus fumigatus, Proteases, MESH: Invasive Pulmonary Aspergillosis, infectious disease, Immunology, MESH: Mice, Inbred BALB C, MESH: Metalloendopeptidases, Microbiology, 03 medical and health sciences, Immune system, MESH: Spores, Fungal, MESH: Lung, Molecular Biology, MESH: Mice, complement system, immune evasion, Innate immune system, MESH: Humans, MESH: Host-Pathogen Interactions, C4A, MESH: Macrophages, Cell Biology, biology.organism_classification, MESH: Male, Complement system, 030104 developmental biology, MESH: Mannose-Binding Protein-Associated Serine Proteases, inflammation, MESH: Disease Models, Animal
Abstract: International audience; Innate immunity in animals including humans encompasses the complement system, which is considered an important host defense mechanism against Aspergillus fumigatus, one of the most ubiquitous opportunistic human fungal pathogens. Previously, it has been shown that the alkaline protease Alp1p secreted from A. fumigatus mycelia degrades the complement components C3, C4, and C5. However, it remains unclear how the fungal spores (i.e. conidia) defend themselves against the activities of the complement system immediately after inhalation into the lung. Here, we show that A. fumigatus conidia contain a metalloprotease Mep1p, which is released upon conidial contact with collagen and inactivates all three complement pathways. In particular, Mep1p efficiently inactivated the major complement components C3, C4, and C5 and their activation products (C3a, C4a, and C5a) as well as the pattern-recognition molecules MBL and ficolin-1, either by directly cleaving them or by cleaving them to a form that is further broken down by other proteases of the complement system. Moreover, incubation of Mep1p with human serum significantly inhibited the complement hemolytic activity and conidial opsonization by C3b and their subsequent phagocytosis by macrophages. Together, these results indicate that Mep1p associated with and released from A. fumigatus conidia likely facilitates early immune evasion by disarming the complement defense in the human host.
Published: 2018

36. The LKB1–AMPK-α1 signaling pathway triggers hypoxic pulmonary vasoconstriction downstream of mitochondria

Author: Marc Foretz, Sandy MacMillan, Benoit Viollet, Javier Moral-Sanz, D. Grahame Hardie, Carmel M. Moran, Sophronia A. Lewis, A. Mark Evans, Fiona A. Ross, Adrian Thomson, Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), College of Life Sciences, and University of Dundee
Subjects: 0301 basic medicine, MESH: Signal Transduction, AMP-Activated Protein Kinases, Mitochondrion, Biochemistry, MESH: Mice, Knockout, MESH: Hypoxia, Hypoxic pulmonary vasoconstriction, Myocyte, MESH: Animals, MESH: AMP-Activated Protein Kinases, Hypoxia, CAMKK2, Mice, Knockout, chemistry.chemical_classification, MESH: Reactive Oxygen Species, MESH: Myocytes, Smooth Muscle, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Mitochondria, 3. Good health, Cell biology, MESH: HEK293 Cells, MESH: Vasoconstriction, medicine.symptom, Signal transduction, Signal Transduction, MESH: Mice, Transgenic, MESH: Mitochondria, Myocytes, Smooth Muscle, MESH: Pulmonary Artery, Mice, Transgenic, Protein Serine-Threonine Kinases, Pulmonary Artery, MESH: Protein-Serine-Threonine Kinases, 03 medical and health sciences, MESH: Mice, Inbred C57BL, medicine, Animals, Humans, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Molecular Biology, Reactive oxygen species, MESH: Humans, AMPK, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell Biology, Hypoxia (medical), Mice, Inbred C57BL, HEK293 Cells, 030104 developmental biology, chemistry, Vasoconstriction, Reactive Oxygen Species
Abstract: International audience; Hypoxic pulmonary vasoconstriction (HPV), which aids ventilation-perfusion matching in the lungs, is triggered by mechanisms intrinsic to pulmonary arterial smooth muscles. The unique sensitivity of these muscles to hypoxia is conferred by mitochondrial cytochrome c oxidase subunit 4 isoform 2, the inhibition of which has been proposed to trigger HPV through increased generation of mitochondrial reactive oxygen species. Contrary to this model, we have shown that the LKB1-AMPK-α1 signaling pathway is critical to HPV. Spectral Doppler ultrasound revealed that deletion of the AMPK-α1 catalytic subunit blocked HPV in mice during mild (8% O2) and severe (5% O2) hypoxia, whereas AMPK-α2 deletion attenuated HPV only during severe hypoxia. By contrast, neither of these genetic manipulations affected serotonin-induced reductions in pulmonary vascular flow. HPV was also attenuated by reduced expression of LKB1, a kinase that activates AMPK during energy stress, but not after deletion of CaMKK2, a kinase that activates AMPK in response to increases in cytoplasmic Ca2+ Fluorescence imaging of acutely isolated pulmonary arterial myocytes revealed that AMPK-α1 or AMPK-α2 deletion did not affect mitochondrial membrane potential during normoxia or hypoxia. However, deletion of AMPK-α1, but not of AMPK-α2, blocked hypoxia from inhibiting KV1.5, the classical "oxygen-sensing" K+ channel in pulmonary arterial myocytes. We conclude that LKB1-AMPK-α1 signaling pathways downstream of mitochondria are critical for the induction of HPV, in a manner also supported by AMPK-α2 during severe hypoxia.
Published: 2018

37. Impaired Kupffer Cell Self-Renewal Alters the Liver Response to Lipid Overload during Non-alcoholic Steatohepatitis

Author: Sébastien Dussaud, Alexandre Boissonnas, Björn E. Clausen, Genevieve Marcelin, Emmanuel L. Gautier, Sophie Tran, Melissa Ouhachi, Lucie Poupel, Ines Baba, Philippe Lesnik, Thierry Huby, Elissa Magréau-Davy, Florian Sennlaub, Laurent Yvan-Charvet, Wilfried Le Goff, Adélaïde Gélineau, Martine Moreau, Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Research Unit on Cardiovascular and Metabolic Diseases [IHU ICAN], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Institut de Cardiométabolisme et Nutrition = Institute of Cardiometabolism and Nutrition [CHU Pitié Salpêtrière] (IHU ICAN), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Nutrition et obésités: approches systémiques (UMR-S 1269) (Nutriomics), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Centre d'Immunologie et des Maladies Infectieuses (CIMI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), University Medical Center of the Johannes Gutenberg-University Mainz, Centre méditerranéen de médecine moléculaire (C3M), Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Côte d'Azur (UCA), FHU OncoAge - Pathologies liées à l’âge [CHU Nice] (OncoAge), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Pharmacologie Moléculaire et Cellulaire [UNIV Côte d'Azur] (UPMC)-Université Côte d'Azur (UCA), Institut de la Vision, Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Institute of cardiometabolism and nutrition (ICAN), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Nutrition et obésités: approches systémiques (nutriomics) (UMR-S 1269 INSERM - Sorbonne Université), Centre d'Immunologie et de Maladies Infectieuses (CIMI), Johannes Gutenberg - Universität Mainz (JGU), Université Nice Sophia Antipolis (... - 2019) (UNS), Programme ATIP - Avenir, Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Research Unit on Cardiovascular and Metabolic Diseases (ICAN), Johannes Gutenberg - Universität Mainz = Johannes Gutenberg University (JGU), and CCSD, Accord Elsevier
Subjects: 0301 basic medicine, [SDV]Life Sciences [q-bio], Ontogeny, MESH: Cell Self Renewal, Self renewal, MESH: Monocytes, MESH: Mice, Knockout, Mice, 0302 clinical medicine, Non-alcoholic Fatty Liver Disease, Immunology and Allergy, Kupffer cells, MESH: Animals, Cell Self Renewal, MESH: Lipid Metabolism, Mice, Knockout, Kupffer cell, Lipids, Research Highlight, macrophages, [SDV] Life Sciences [q-bio], Infectious Diseases, medicine.anatomical_structure, Liver, 030220 oncology & carcinogenesis, monocytes, medicine.medical_specialty, non-alcoholic steatohepatitis (NASH), Immunology, Biology, 03 medical and health sciences, MESH: Mice, Inbred C57BL, MESH: Cell Proliferation, Internal medicine, medicine, Animals, Liver damage, MESH: Mice, Cell Proliferation, MESH: Non-alcoholic Fatty Liver Disease, Triglyceride storage, Non alcoholic, Lipid Metabolism, medicine.disease, MESH: Lipids, eye diseases, Mice, Inbred C57BL, MESH: Kupffer Cells, 030104 developmental biology, Endocrinology, Steatohepatitis, Homeostasis, MESH: Liver
Abstract: International audience; Kupffer cells (KCs) are liver-resident macrophages that self-renew by proliferation in the adult independently from monocytes. However, how they are maintained during non-alcoholic steatohepatitis (NASH) remains ill defined. We found that a fraction of KCs derived from Ly-6C+ monocytes during NASH, underlying impaired KC self-renewal. Monocyte-derived KCs (MoKCs) gradually seeded the KC pool as disease progressed in a response to embryo-derived KC (EmKC) death. Those MoKCs were partly immature and exhibited a pro-inflammatory status compared to EmKCs. Yet, they engrafted the KC pool for the long term as they remained following disease regression while acquiring mature EmKC markers. While KCs as a whole favored hepatic triglyceride storage during NASH, EmKCs promoted it more efficiently than MoKCs, and the latter exacerbated liver damage, highlighting functional differences among KCs with different origins. Overall, our data reveal that KC homeostasis is impaired during NASH, altering the liver response to lipids, as well as KC ontogeny.
Published: 2020

38. A Quinoline-Based DNA Methyltransferase Inhibitor as a Possible Adjuvant in Osteosarcoma Therapy

Author: Roberta Mazzone, Piero Picci, Katia Scotlandi, Michela Pasello, Patrizia Nanni, Giordano Nicoletti, Maria Cristina Manara, Antonello Mai, Clara Guerzoni, Camilla Cristalli, Pier Luigi Lollini, Lorena Landuzzi, Sergio Valente, Giulia Stazi, Paola B. Arimondo, Clemens Zwergel, Istituto Ortopedico Rizzoli [Bologna, Italy], Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], Centre National de la Recherche Scientifique (CNRS), Department of Medicinal Chemistry and Technologies, Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome], This work was funded by AIRC (IG 18451 to K. Scotlandi, IG 19162 to A. Mai), the Italian Ministry of Health (TRANSCAN_ project TORPEDO_ER-2015-23604059 to K. Scotlandi), Progetto Ateneo Sapienza (to A. Mai), PRIN 2016 (prot 20152TE5PK to A. Mai), and the NIH (R01GM114306 to A. Mai)., The authors thank Cristina Ghinelli for editing the manuscript., Manara, Maria Cristina, Valente, Sergio, Cristalli, Camilla, Nicoletti, Giordano, Landuzzi, Lorena, Zwergel, Clemen, Mazzone, Roberta, Stazi, Giulia, Arimondo, Paola B, Pasello, Michela, Guerzoni, Clara, Picci, Piero, Nanni, Patrizia, Lollini, Pier-Luigi, Mai, Antonello, and Scotlandi, Katia
Subjects: 0301 basic medicine, Male, patient-derived xenograft (PDX), MESH: Osteosarcoma/genetics, MESH: Antineoplastic Combined Chemotherapy Protocols/therapeutic use, Cellular differentiation, DNA Methyltransferase Inhibitor, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, chemotherapy, MESH: Mice, Knockout, 0302 clinical medicine, MESH: Osteosarcoma/drug therapy, Antineoplastic Combined Chemotherapy Protocols, MESH: Aminoquinolines/administration & dosage, MESH: Animals, MESH: Quinolines/chemistry, Enzyme Inhibitors, MESH: DNA (Cytosine-5-)-Methyltransferase 1/metabolism, Mice, Knockout, DNA methylation, Chemistry, 3. Good health, Tumor Burden, MESH: Benzamides/pharmacology, Gene Expression Regulation, Neoplastic, 030220 oncology & carcinogenesis, Benzamides, oncology, Aminoquinolines, Quinolines, MESH: Cisplatin/administration & dosage, MESH: Bone Neoplasms/metabolism, Osteosarcoma, MESH: Doxorubicin/administration & dosage, epigenetic, MESH: Bone Neoplasms/genetics, medicine.drug, DNA (Cytosine-5-)-Methyltransferase 1, musculoskeletal diseases, MESH: Aminoquinolines/pharmacology, MESH: Cell Line, Tumor, DNA damage, MESH: Gene Expression Regulation, Neoplastic/drug effects, MESH: Tumor Burden/genetics, MESH: Xenograft Model Antitumor Assays, Bone Neoplasms, [SDV.CAN]Life Sciences [q-bio]/Cancer, 03 medical and health sciences, Cell Line, Tumor, osteosarcoma, medicine, Animals, Humans, Doxorubicin, MESH: DNA (Cytosine-5-)-Methyltransferase 1/antagonists & inhibitors, MESH: Enzyme Inhibitors/chemistry, Cisplatin, MESH: Humans, Nucleoside inhibitor, medicine.disease, MESH: Enzyme Inhibitors/pharmacology, Xenograft Model Antitumor Assays, MESH: Male, MESH: Enzyme Inhibitors/administration & dosage, MESH: Bone Neoplasms/drug therapy, 030104 developmental biology, MESH: Osteosarcoma/metabolism, Cancer research, cancer research, MESH: DNA (Cytosine-5-)-Methyltransferase 1/genetics, MESH: Benzamides/administration & dosage, MESH: Tumor Burden/drug effects
Abstract: The identification of new therapeutic strategies against osteosarcoma, the most common primary bone tumor, continues to be a primary goal to improve the outcomes of patients refractory to conventional chemotherapy. Osteosarcoma originates from the transformation of mesenchymal stem cells (MSC) and/or osteoblast progenitors, and the loss of differentiation is a common biological osteosarcoma feature, which has strong significance in predicting tumor aggressiveness. Thus, restoring differentiation through epigenetic reprogramming is potentially exploitable for therapeutic benefits. Here, we demonstrated that the novel nonnucleoside DNMT inhibitor (DNMTi) MC3343 affected tumor proliferation by blocking osteosarcoma cells in G1 or G2–M phases and induced osteoblastic differentiation through the specific reexpression of genes regulating this physiologic process. Although MC3343 has a similar antiproliferative effect as 5azadC, the conventional FDA-approved nucleoside inhibitor of DNA methylation, its effects on cell differentiation are distinct. Induction of the mature osteoblast phenotype coupled with a sustained cytostatic response was also confirmed in vivo when MC3343 was used against a patient-derived xenograft (PDX). In addition, MC3343 displayed synergistic effects with doxorubicin and cisplatin (CDDP), two major chemotherapeutic agents used to treat osteosarcoma. Specifically, MC3343 increased stable doxorubicin bonds to DNA, and combined treatment resulted in sustained DNA damage and increased cell death. Overall, this nonnucleoside DNMTi is an effective novel agent and is thus a potential therapeutic option for patients with osteosarcoma who respond poorly to preadjuvant chemotherapy. Mol Cancer Ther; 17(9); 1881–92. ©2018 AACR.
Published: 2018

39. The Protozoan Parasite Toxoplasma gondii Selectively Reprograms the Host Cell Translatome

Author: Julie Lorent, Maritza Jaramillo, Laia Masvidal, Tommy Alain, Ola Larsson, Visnu Chaparro, Bruno D. Fonseca, Tyson E. Graber, Léon C van Kempen, Maria Aguirre, Louis-Philippe Leroux, Institut Armand Frappier (INRS-IAF), Réseau International des Instituts Pasteur (RIIP)-Institut National de la Recherche Scientifique [Québec] (INRS), Karolinska Institutet [Stockholm], University of Ottawa [Ottawa], Jewish General Hospital, McGill University = Université McGill [Montréal, Canada], University Medical Center Groningen [Groningen] (UMCG), This work was supported by a Basil O'Connor starter scholar research award (5-FY14-78) and a research grant (6-FY16-151) from The March of Dimes Foundation to M.J. The Centre for Host-Parasite Interactions is supported by a Subvention de Regroupement Stratégique from the Fonds de Recherche du Québec en Nature et Technologies (FRQ-NT). M.J. is a recipient of a Bourse de Chercheur-Boursier Junior 1 from the Fonds de Recherche du Québec en Santé (FRQ-S) and a Subvention d'Établissement de Jeune Chercheur from the FRQ-S. V.C. is supported by a Ph.D. scholarship from the Fondation Universitaire Armand Frappier. Research in the laboratory of O.L. is supported by grants from the Swedish Research Council and the Wallenberg Academy Fellows program., and We are grateful to Nahum Sonenberg for providing bone marrow from C57BL/6 s6k1−/− s6k2−/− mice (McGill University, Montreal, QC, Canada). We thank Medhi Jafarnejad (McGill University) for technical advice. We are thankful to Annie Sylvestre and Annik Lafrance for invaluable technical assistance. We acknowledge support from the Science for Life Laboratory, the National Genomics Infrastructure (NGI), and Uppmax for providing assistance in massive parallel sequencing and computational infrastructure.
Subjects: MESH: Signal Transduction, 0301 basic medicine, DIFFERENTIAL TRANSLATION, Protozoan Proteins, MESH: Toxoplasma/pathogenicity, RNA 5' Terminal Oligopyrimidine Sequence, MESH: Mice, Knockout, Mice, Poly(A)-binding protein, Protein biosynthesis, host-pathogen interactions, MESH: Animals, Cells, Cultured, Mice, Knockout, biology, IMMUNE-RESPONSES, TOR Serine-Threonine Kinases, translational control, Translation (biology), Cell biology, macrophages, Infectious Diseases, MESSENGER-RNA TRANSLATION, mTOR, PROTEIN-SYNTHESIS, MESH: Host-Parasite Interactions, [SDV.IMM]Life Sciences [q-bio]/Immunology, Toxoplasma, Signal Transduction, MESH: Cells, Cultured, Translational efficiency, Immunology, ENDOPLASMIC-RETICULUM, MESH: RNA 5' Terminal Oligopyrimidine Sequence, Toxoplasma gondii, Microbiology, DENDRITIC CELLS, Host-Parasite Interactions, POLY(A)-BINDING PROTEINS, Mitochondrial Proteins, 03 medical and health sciences, MESH: Protein Biosynthesis/genetics, MESH: Mice, Inbred C57BL, PARASITOPHOROUS VACUOLE MEMBRANE, parasitic diseases, Animals, MESH: Mice, Mechanistic target of rapamycin, Cellular Microbiology: Pathogen-Host Cell Molecular Interactions, 030102 biochemistry & molecular biology, Intracellular parasite, biology.organism_classification, MESH: Protozoan Proteins/immunology, Cytoplasm organization, Mice, Inbred C57BL, MESH: Mitochondrial Proteins/genetics, MAMMALIAN-TARGET, 030104 developmental biology, Protein Biosynthesis, MESH: TOR Serine-Threonine Kinases/genetics, biology.protein, INNATE IMMUNITY, MESH: Macrophages/parasitology, Parasitology
Abstract: International audience; The intracellular parasite Toxoplasma gondii promotes infection by targeting multiple host cell processes; however, whether it modulates mRNA translation is currently unknown. Here, we show that infection of primary murine macrophages with type I or II T. gondii strains causes a profound perturbation of the host cell translatome. Notably, translation of transcripts encoding proteins involved in metabolic activity and components of the translation machinery was activated upon infection. In contrast, the translational efficiency of mRNAs related to immune cell activation and cytoskeleton/cytoplasm organization was largely suppressed. Mechanistically, T. gondii bolstered mechanistic target of rapamycin (mTOR) signaling to selectively activate the translation of mTOR-sensitive mRNAs, including those with a 5'-terminal oligopyrimidine (5' TOP) motif and those encoding mitochondrion-related proteins. Consistent with parasite modulation of host mTOR-sensitive translation to promote infection, inhibition of mTOR activity suppressed T. gondii replication. Thus, selective reprogramming of host mRNA translation represents an important subversion strategy during T. gondii infection.
Published: 2018

40. Deficient LRRC8A-dependent volume-regulated anion channel activity is associated with male infertility in mice

Author: Taiping Chen, John H. Richburg, Sharon Y.R. Dent, Raif S. Geha, Jean Louis Guénet, Caitlin J. Murphy, Carlos J. Perez, Janet Chou, Jean Jaubert, Tewfik Hamidi, Meichun Deng, Craig D. Platt, Qinghua Shi, Yue Lu, Héctor Gaitán-Peñas, Fernando Benavides, Kevin Lin, Jeesun Kim, Hui Lin Pan, Mark T. Bedford, Jianqiang Bao, Meng Hua Zhou, Huan Zhang, Yuying Huang, Raúl Estévez, The University of Texas M.D. Anderson Cancer Center [Houston], School of Life Science [Heifei], University of Science and Technology of China [Hefei] (USTC), University of Texas at Austin [Austin], Génétique de la souris - Mouse Genetics, Institut Pasteur [Paris], Boston Children's Hospital, Harvard Medical School [Boston] (HMS), Center for Neuroscience & Pain Research [Houston], Institut d'Investigació Biomèdica de Bellvitge [Barcelone] (IDIBELL), CIBER de Enfermedades Raras (CIBERER), Graduate School of Biomedical Sciences [Houston], The University of Texas Health Science Center at Houston (UTHealth)-The University of Texas M.D. Anderson Cancer Center [Houston], This study made use of the Research Animal Support Facility-Smithville (including Laboratory Animal Genetic Services and Research Histology Pathology and Imaging Core), the Sequencing and Microarray Facility, and the HREMF, all of which are supported by P30 CA016672 DHHS/NCI Cancer Center Support Grant to MD Anderson Cancer Center. This work was supported in part by Departmental Funds and Molecular Biology Facility Core supported by CPRIT grant RP170002 to Dr. Jianjun Shen, National Basic Research Program of China (2014CB943101) and the National Natural Science Foundation of China (31371519 and 313111245) to QS, and R01 ES016591 to JHR., We thank the Research Animal Support Facility-Smithville for their assistance with the maintenance of the mouse strains, the Histology Core for processing the samples, the Molecular Biology Facility Core for RNA sequencing, the Bioinformatics and Biostatistics Services for data analysis, and Briana Dennehey for editing the manuscript., and Institut Pasteur [Paris] (IP)
Subjects: Male, 0301 basic medicine, MESH: Membrane Proteins / genetics, medicine.disease_cause, MESH: Mice, Knockout, Male infertility, Mice, MESH: Testis / pathology, Testis, MESH: Animals, MESH: Membrane Proteins / metabolism, MESH: Healthy Volunteers, MESH: Infertility, Male / diagnosis, Mice, Knockout, Reproductive Biology, Mutation, biology, Esterilitat en els animals, General Medicine, Anion channel activity, MESH: Biological Transport, Active / genetics, Spermatozoa, MESH: Case-Control Studies, Healthy Volunteers, MESH: China, MESH: Spermatozoa / cytology, MESH: Biomarkers / analysis, Cell biology, medicine.anatomical_structure, Flagella, MESH: Cell Survival / genetics, Ion channels, Sperm Motility, Female, Germ cell, Research Article, Adult, Anions, China, endocrine system, MESH: Mutation, Cell Survival, Sterility, Biological Transport, Active, MESH: Spermatozoa / pathology, 03 medical and health sciences, Infertility in animals, Genetics, medicine, Animals, Humans, MESH: Mice, Infertility, Male, MESH: Ion Transport / genetics, MESH: Infertility, Male / pathology, Ion Transport, MESH: Humans, MESH: Sperm Motility / genetics, Membrane Proteins, MESH: Adult, medicine.disease, Sperm, MESH: Male, MESH: Anions / metabolism, Disease Models, Animal, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, Insulin receptor, Fertility, MESH: Flagella / pathology, 030104 developmental biology, Apoptosis, Case-Control Studies, biology.protein, MESH: Disease Models, Animal, MESH: Infertility, Male / genetics, MESH: Female, Biomarkers
Abstract: International audience; Ion channel-controlled cell volume regulation is of fundamental significance to the physiological function of sperm. In addition to volume regulation, LRRC8A-dependent volume-regulated anion channel (VRAC) activity is involved in cell cycle progression, insulin signaling, and cisplatin resistance. Nevertheless, the contribution of LRRC8A and its dependent VRAC activity in the germ cell lineage remain unknown. By utilizing a spontaneous Lrrc8a mouse mutation (c.1325delTG, p.F443*) and genetically engineered mouse models, we demonstrate that LRRC8A-dependent VRAC activity is essential for male germ cell development and fertility. Lrrc8a-null male germ cells undergo progressive degeneration independent of the apoptotic pathway during postnatal testicular development. Lrrc8a-deficient mouse sperm exhibit multiple morphological abnormalities of the flagella (MMAF), a feature commonly observed in the sperm of infertile human patients. Importantly, we identified a human patient with a rare LRRC8A hypomorphic mutation (c.1634G>A, p.Arg545His) possibly linked to Sertoli cell-only syndrome (SCOS), a male sterility disorder characterized by the loss of germ cells. Thus, LRRC8A is a critical factor required for germ cell development and volume regulation in the mouse, and it might serve as a novel diagnostic and therapeutic target for SCOS patients.
Published: 2018

41. Metformin directly targets the H3K27me3 demethylase KDM6A/UTX

Author: Salvador Fernández-Arroyo, Lukas Werner, Jorge Joven, Laura Llorach-Parés, Javier A. Menendez, Noemí Cabré, Benoit Viollet, Sara Verdura, Elisabet Cuyàs, Fedra Luciano-Mateo, Jan Stursa, Alfons Nonell-Canals, Jiri Neuzil, Begoña Martin-Castillo, Melchor Sanchez-Martinez, Program Against Cancer Therapeutic Resistance/Metabolism & Cancer Group [Catalonia, Spain] (ProCURE), Catalan Institute of Oncology-Girona (ICO-Girona), Girona Biomedical Research Institute [Girona, Spain] (IDIBGI), Mind the Byte [Barcelona, Spain], Unitat de Recerca Biomedica [Reus, Spain], Hospital Universitari de Sant Joan [Reus, Spain]-Rovira i Virgili University [Reus, Spain] (IISPV), Institute of Chemical Technology [Prague, Czech Republic], Institute of Biotechnology [Prague-West, Czech Republic], Czech Academy of Sciences [Prague] (CAS), Girona Biomedical Research Institute (IdIBGi), Institut d'Investigació Biomèdica de Girona Dr. Josep Trueta - Girona Biomedical Research Institute (IDIBGI), [Institut Cochin] Département Endocrinologie, métabolisme, diabète (EMD) (EMD), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Menzies Health Institute Queensland [Queensland, Australia], Griffith University [Brisbane], This work was supported by grants from the Ministerio de Ciencia e Innovacion (Grant SAF2016-80639-P to J. A. Menendez), Plan Nacional de I+D+I, Spain, the Agencia de Gestio d’Ajuts Universitaris i de Recerca (AGAUR) (Grant 2014 SGR229 to J. A. Menendez), Departament d’Economia i Coneixement, Catalonia, Spain, and the Czech Health Council Foundation (Grant 16- 31604A to J. Neuzil). Elisabet Cuyas is supported by the Sara Borrell postdoctoral contract (CD15/00033) from the Ministerio de Sanidad y Consumo, Fondo de Investigacion Sanitaria (FIS), Spain. This study was supported also by unrestricted research grants from Roche Pharma (Spain) and Astellas Pharma (Spain) to the Program Against Cancer Therapeutic Resistance (ProCURE, Catalan Institute of Oncology), and the Armangue family (Girona, Catalonia) to the Metabolism and Cancer Group (Girona Biomedical Research Institute), and Viollet, Benoit
Subjects: 0301 basic medicine, Models, Molecular, Aging, Pharmacology, Ligands, MESH: Metformin, MESH: Mice, Knockout, computational screening, MESH: Dose-Response Relationship, Drug, Mice, MESH: Ligands, MESH: Animals, Enzyme Inhibitors, Càncer, Cancer, [SDV.MHEP.EM] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Histone Demethylases, Mice, Knockout, Molecular Structure, Nuclear Proteins, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, 3. Good health, Metformin, Chromatin, MESH: Neoplasms, Experimental, MESH: Enzyme Inhibitors, Original Article, Female, MESH: Biocatalysis, MESH: Models, Molecular, medicine.drug, Premature aging, MESH: Molecular Structure, macromolecular substances, Biology, chemoinformatics, 03 medical and health sciences, Envelliment, MESH: Histone Demethylases, medicine, Animals, Humans, cancer, Epigenetics, MESH: Mice, MESH: Humans, Dose-Response Relationship, Drug, aging, AMPK, Cell Biology, Epigenome, Original Articles, Neoplasms, Experimental, medicine.disease, 030104 developmental biology, biology.protein, Biocatalysis, Demethylase, metformin, MESH: Nuclear Proteins, MESH: Female
Abstract: International audience; Metformin, the first drug chosen to be tested in a clinical trial aimed to target the biology of aging per se, has been clinically exploited for decades in the absence of a complete understanding of its therapeutic targets or chemical determinants. We here outline a systematic chemoinformatics approach to computationally predict biomolecular targets of metformin. Using several structure- and ligand-based software tools and reference databases containing 1,300,000 chemical compounds and more than 9,000 binding sites protein cavities, we identified 41 putative metformin targets including several epigenetic modifiers such as the member of the H3K27me3-specific demethylase subfamily, KDM6A/UTX. AlphaScreen and AlphaLISA assays confirmed the ability of metformin to inhibit the demethylation activity of purified KDM6A/UTX enzyme. Structural studies revealed that metformin might occupy the same set of residues involved in H3K27me3 binding and demethylation within the catalytic pocket of KDM6A/UTX. Millimolar metformin augmented global levels of H3K27me3 in cultured cells, including reversion of global loss of H3K27me3 occurring in premature aging syndromes, irrespective of mitochondrial complex I or AMPK. Pharmacological doses of metformin in drinking water or intraperitoneal injection significantly elevated the global levels of H3K27me3 in the hepatic tissue of low-density lipoprotein receptor-deficient mice and in the tumor tissues of highly aggressive breast cancer xenograft-bearing mice. Moreover, nondiabetic breast cancer patients receiving oral metformin in addition to standard therapy presented an elevated level of circulating H3K27me3. Our biocomputational approach coupled to experimental validation reveals that metformin might directly regulate the biological machinery of aging by targeting core chromatin modifiers of the epigenome.
Published: 2018

42. The VAMP‐associated protein VAPB is required for cardiac and neuronal pacemaker channel function

Author: Nicole, Silbernagel, Magdalena, Walecki, Martin K-H, Schäfer, Mirjam, Kessler, Mehrnoush, Zobeiri, Susanne, Rinné, Aytug K, Kiper, Marlene A, Komadowski, Kirsty S, Vowinkel, Konstantin, Wemhöner, Lisa, Fortmüller, Marcus, Schewe, Amalia M, Dolga, Jelena, Scekic-Zahirovic, Lina A, Matschke, Carsten, Culmsee, Thomas, Baukrowitz, Laurent, Monassier, Nina D, Ullrich, Luc, Dupuis, Steffen, Just, Thomas, Budde, Larissa, Fabritz, Niels, Decher, Phillips University [Marburg, Germany] (PU), Universitätsklinikum Ulm - University Hospital of Ulm, University of Münster, University Hospital Münster - Universitaetsklinikum Muenster [Germany] (UKM), Christian-Albrechts University of Kiel, Laboratoire de pharmacologie et de toxicologie neurocardiovasculaire (LPTNC), Université de Strasbourg (UNISTRA), Universität Heidelberg [Heidelberg], Mécanismes Centraux et Périphériques de la Neurodégénérescence, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), University Hospitals Birmingham [Birmingham, Royaume-Uni], University of Birmingham [Birmingham], Westfälische Wilhelms-Universität Münster = University of Münster (WWU), and Dieterle, Stéphane
Subjects: Pacemaker, Artificial, Embryo, Nonmammalian, MESH: Vesicular Transport Proteins, MESH: Rats, [SDV]Life Sciences [q-bio], Vesicular Transport Proteins, MESH: Neurons, MESH: Carrier Proteins, MESH: Rats, Sprague-Dawley, MESH: Mice, Knockout, Rats, Sprague-Dawley, HCN channels, Mice, Xenopus laevis, cardiac arrhythmia, MESH: Xenopus laevis, Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels, Animals, Humans, MESH: Animals, MESH: Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels, MESH: Zebrafish, MESH: Mice, Zebrafish, Mice, Knockout, Neurons, MESH: Humans, Research, Membrane Proteins, MESH: Embryo, Nonmammalian, Heart, MESH: Ion Channel Gating, Rats, [SDV] Life Sciences [q-bio], MESH: Heart, MESH: HeLa Cells, MESH: Pacemaker, Artificial, Female, MESH: Membrane Proteins, ALS, Carrier Proteins, Ion Channel Gating, MESH: Female, HeLa Cells
Abstract: International audience; Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels encode neuronal and cardiac pacemaker currents. The composition of pacemaker channel complexes in different tissues is poorly understood, and the presence of additional HCN modulating subunits was speculated. Here we show that vesicle-associated membrane protein-associated protein B (VAPB), previously associated with a familial form of amyotrophic lateral sclerosis 8, is an essential HCN1 and HCN2 modulator. VAPB significantly increases HCN2 currents and surface expression and has a major influence on the dendritic neuronal distribution of HCN2. Severe cardiac bradycardias in VAPB-deficient zebrafish and VAPB-/- mice highlight that VAPB physiologically serves to increase cardiac pacemaker currents. An altered T-wave morphology observed in the ECGs of VAPB-/- mice supports the recently proposed role of HCN channels for ventricular repolarization. The critical function of VAPB in native pacemaker channel complexes will be relevant for our understanding of cardiac arrhythmias and epilepsies, and provides an unexpected link between these diseases and amyotrophic lateral sclerosis.-Silbernagel, N., Walecki, M., Schäfer, M.-K. H., Kessler, M., Zobeiri, M., Rinné, S., Kiper, A. K., Komadowski, M. A., Vowinkel, K. S., Wemhöner, K., Fortmüller, L., Schewe, M., Dolga, A. M., Scekic-Zahirovic, J., Matschke, L. A., Culmsee, C., Baukrowitz, T., Monassier, L., Ullrich, N. D., Dupuis, L., Just, S., Budde, T., Fabritz, L., Decher, N. The VAMP-associated protein VAPB is required for cardiac and neuronal pacemaker channel function.
Published: 2018

43. NMDA-Type Glutamate Receptor Activation Promotes Vascular Remodeling and Pulmonary Arterial Hypertension

Author: Elie Fadel, Morad K. Nakhleh, Audrey Courboulin, Marceau Quatredeniers, Catherine Rucker-Martin, Sylvia Cohen-Kaminsky, Maria-Candida Vinhas, Sébastien J. Dumas, Peter Dorfmüller, Marc Humbert, Gilles Bru-Mercier, Nicolas Raymond, Fabrice Antigny, Benoit Ranchoux, Matthieu Vocelle, Frédéric Perros, Elodie Gouadon, Hypertension arterielle pulmonaire physiopathologie et innovation thérapeutique, Centre chirurgical Marie Lannelongue-Institut National de la Santé et de la Recherche Médicale (INSERM), Remodelage tissulaire et fonctionnel : signalisation et physio-pathologie (RTFSPP), Centre National de la Recherche Scientifique (CNRS)-Centre chirurgical Marie Lannelongue-Université Paris-Sud - Paris 11 (UP11), Centre chirurgical Marie Lannelongue, Inserm, UMR_S 999 – LabEx LERMIT, Centre Chirurgical Marie-Lannelongue, Le Plessis-Robinson 5Université Paris-Sud, Faculté de Médecine, Le Kremlin-Bicêtre, France, and Université Paris-Saclay
Subjects: 0301 basic medicine, MESH: Signal Transduction, MESH: Endothelin-1, Platelet-derived growth factor, [SDV]Life Sciences [q-bio], Apoptosis, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], 030204 cardiovascular system & hematology, Pharmacology, MESH: Mice, Knockout, [SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, Muscle, Smooth, Vascular, platelet-derived growth factor, chemistry.chemical_compound, Mice, 0302 clinical medicine, pulmonary arterial hypertension, Medicine, MESH: Animals, Endothelial dysfunction, Lung, Mice, Knockout, Receptors, Endothelin, musculoskeletal, neural, and ocular physiology, Glutamate receptor, MESH: Glutamic Acid, MESH: Muscle, Smooth, Vascular, 3. Good health, Potassium Channels, Voltage-Gated, MESH: Calcium, endothelin-1, NMDA receptor, Cardiology and Cardiovascular Medicine, Endothelin receptor, MESH: Vascular Remodeling, Signal Transduction, MESH: Rats, Hypertension, Pulmonary, vascular remodeling, Glutamic Acid, glutamate, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Receptors, N-Methyl-D-Aspartate, 03 medical and health sciences, Growth factor receptor, MESH: Receptors, Endothelin, Physiology (medical), MESH: Cell Proliferation, mental disorders, Animals, Humans, MESH: Lung, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Mice, smooth muscle cell, Cell Proliferation, MESH: Receptors, N-Methyl-D-Aspartate, MESH: Humans, MESH: Hypertension, Pulmonary, business.industry, MESH: Apoptosis, MESH: Dizocilpine Maleate, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, medicine.disease, Pulmonary hypertension, Endothelin 1, Rats, Disease Models, Animal, 030104 developmental biology, chemistry, nervous system, Calcium, Dizocilpine Maleate, MESH: Disease Models, Animal, business, MESH: Potassium Channels, Voltage-Gated, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: Background: Excessive proliferation and apoptosis resistance in pulmonary vascular cells underlie vascular remodeling in pulmonary arterial hypertension (PAH). Specific treatments for PAH exist, mostly targeting endothelial dysfunction, but high pulmonary arterial pressure still causes heart failure and death. Pulmonary vascular remodeling may be driven by metabolic reprogramming of vascular cells to increase glutaminolysis and glutamate production. The N -methyl- d -aspartate receptor (NMDAR), a major neuronal glutamate receptor, is also expressed on vascular cells, but its role in PAH is unknown. Methods: We assessed the status of the glutamate-NMDAR axis in the pulmonary arteries of patients with PAH and controls through mass spectrometry imaging, Western blotting, and immunohistochemistry. We measured the glutamate release from cultured pulmonary vascular cells using enzymatic assays and analyzed NMDAR regulation/phosphorylation through Western blot experiments. The effect of NMDAR blockade on human pulmonary arterial smooth muscle cell proliferation was determined using a BrdU incorporation assay. We assessed the role of NMDARs in vascular remodeling associated to pulmonary hypertension, in both smooth muscle-specific NMDAR knockout mice exposed to chronic hypoxia and the monocrotaline rat model of pulmonary hypertension using NMDAR blockers. Results: We report glutamate accumulation, upregulation of the NMDAR, and NMDAR engagement reflected by increases in GluN1-subunit phosphorylation in the pulmonary arteries of human patients with PAH. K v channel inhibition and type A-selective endothelin receptor activation amplified calcium-dependent glutamate release from human pulmonary arterial smooth muscle cell, and type A-selective endothelin receptor and platelet-derived growth factor receptor activation led to NMDAR engagement, highlighting crosstalk between the glutamate-NMDAR axis and major PAH-associated pathways. The platelet-derived growth factor-BB-induced proliferation of human pulmonary arterial smooth muscle cells involved NMDAR activation and phosphorylated GluN1 subunit localization to cell-cell contacts, consistent with glutamatergic communication between proliferating human pulmonary arterial smooth muscle cells via NMDARs. Smooth-muscle NMDAR deficiency in mice attenuated the vascular remodeling triggered by chronic hypoxia, highlighting the role of vascular NMDARs in pulmonary hypertension. Pharmacological NMDAR blockade in the monocrotaline rat model of pulmonary hypertension had beneficial effects on cardiac and vascular remodeling, decreasing endothelial dysfunction, cell proliferation, and apoptosis resistance while disrupting the glutamate-NMDAR pathway in pulmonary arteries. Conclusions: These results reveal a dysregulation of the glutamate-NMDAR axis in the pulmonary arteries of patients with PAH and identify vascular NMDARs as targets for antiremodeling treatments in PAH.
Published: 2018

44. Rescue of GSDIII Phenotype with Gene Transfer Requires Liver- and Muscle-Targeted GDE Expression

Author: Federico Mingozzi, Patrice Vidal, Helena Costa Verdera, Bernard Gjata, Francesco Puzzo, Giuseppe Ronzitti, Laetitia van Wittenberghe, Giacomo P. Comi, Gilles Mithieux, Fanny Collaud, Sabrina Lucchiari, Pascal Laforêt, Edoardo Malfatti, Louisa Jauze, Monika Gjorgjieva, Fabienne Rajas, Marcelo Simon Sola, Solenne Marmier, Alban Vignaud, Serena Pagliarani, Severine Charles, Isabelle Richard, Pasqualina Colella, Université Pierre et Marie Curie - Paris 6 (UPMC), Approches génétiques intégrées et nouvelles thérapies pour les maladies rares (INTEGRARE), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Généthon, Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Physiopatology and Transplantation, University of Milan (DEPT), Università degli Studi di Milano = University of Milan (UNIMI), IRCCS Foundation Ca' Granda Ospedale Maggiore Policlinico, Généthon, Université Pierre et Marie Curie - Paris 6 - UFR de Médecine Pierre et Marie Curie (UPMC), Nutrition et cerveau (U1213, U855), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Department of Pathophysiology and Transplantation, Institut de Myologie, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), U855, Nutrition et cerveau, Centre de recherche en Myologie – U974 SU-INSERM, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), ANR-17-CE18-0014,TRACeGSDIII,Optimisation translationnelle d'un vecteur AAV pour le traitement de GSDIII(2017), École pratique des hautes études (EPHE), Richard, Isabelle, Centre de recherche en myologie, Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut national de la santé et de la recherche médicale, Université d’Evry Val d’Essonne, Genethon (INSERM), University of Milan, Université de Lyon, Généthon, Evry, Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Association française contre les myopathies (AFM-Téléthon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Nutrition, diabète et cerveau, École pratique des hautes études (EPHE)-Université d'Évry-Val-d'Essonne (UEVE)-GENETHON 3-Institut National de la Santé et de la Recherche Médicale (INSERM), GENETHON, Genethon, Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Généthon-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), and Centre National de la Recherche Scientifique (CNRS)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Association française contre les myopathies (AFM-Téléthon)-Sorbonne Université (SU)
Subjects: Blood Glucose, Male, 0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Genetic enhancement, MESH: Glycogen, Gene Expression, MESH: Dependovirus, Glycogen storage disease type III, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH: Mice, Knockout, MESH: Hepatocytes, Mice, chemistry.chemical_compound, dual AAV vectors, MESH: Genetic Vectors, Drug Discovery, MESH: Animals, Vector (molecular biology), MESH: Glycogen Storage Disease Type III, MESH: Organ Specificity, Mice, Knockout, MESH: Muscle, Skeletal, Glycogen, Gene Transfer Techniques, Dependovirus, Phenotype, Cori disease, gene therapy, 3. Good health, Cell biology, Liver, Organ Specificity, Acid alpha-glucosidase, Molecular Medicine, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Original Article, medicine.symptom, MESH: Enzyme Activation, MESH: Gene Expression, Transgene, Genetic Vectors, adeno-associated vector, MESH: Gene Transfer Techniques, Biology, [SDV.GEN.GH] Life Sciences [q-bio]/Genetics/Human genetics, MESH: Phenotype, 03 medical and health sciences, glycogenosis, Genetics, medicine, Animals, MESH: Glycogen Debranching Enzyme System, Muscle, Skeletal, Molecular Biology, MESH: Mice, glycogen storage disease type III, Pharmacology, MESH: Genetic Therapy, Muscle weakness, Glycogen Debranching Enzyme System, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Genetic Therapy, neuromuscular disease, medicine.disease, MESH: Male, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Enzyme Activation, Disease Models, Animal, 030104 developmental biology, chemistry, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, acid-alpha-glucosidase, Hepatocytes, MESH: Biomarkers, MESH: Blood Glucose, MESH: Disease Models, Animal, Biomarkers, MESH: Liver
Abstract: International audience; Glycogen storage disease type III (GSDIII) is an autosomal recessive disorder caused by a deficiency of glycogen-debranching enzyme (GDE), which results in profound liver metabolism impairment and muscle weakness. To date, no cure is available for GSDIII and current treatments are mostly based on diet. Here we describe the development of a mouse model of GSDIII, which faithfully recapitulates the main features of the human condition. We used this model to develop and test novel therapies based on adeno-associated virus (AAV) vector-mediated gene transfer. First, we showed that overexpression of the lysosomal enzyme alpha-acid glucosidase (GAA) with an AAV vector led to a decrease in liver glycogen content but failed to reverse the disease phenotype. Using dual overlapping AAV vectors expressing the GDE transgene in muscle, we showed functional rescue with no impact on glucose metabolism. Liver expression of GDE, conversely, had a direct impact on blood glucose levels. These results provide proof of concept of correction of GSDIII with AAV vectors, and they indicate that restoration of the enzyme deficiency in muscle and liver is necessary to address both the metabolic and neuromuscular manifestations of the disease.
Published: 2018

45. Transcriptomic Analysis of Two Cdsn-Deficient Mice Shows Gene Signatures Biologically Relevant for Peeling Skin Disease

Author: F. Valentin, Juliette Mazereeuw-Hautier, Nathalie Jonca, Guy Serre, Anne Huchenq, M. Pichery, Vinzenz Oji, Sarra Zaafouri, CARBILLET, Véronique, Unité différenciation épidermique et auto-immunité rhumatoïde (UDEAR), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), University Hospital Münster - Universitaetsklinikum Muenster [Germany] (UKM), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), and CHU Toulouse [Toulouse]
Subjects: 0301 basic medicine, Disease, Biochemistry, MESH: Mice, Knockout, MESH: Down-Regulation, MESH: Dermatitis, Exfoliative / pathology, Transcriptome, Mice, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Deficient mouse, MESH: Up-Regulation, MESH: Animals, MESH: Skin Diseases, Genetic / pathology, ComputingMilieux_MISCELLANEOUS, Mice, Knockout, MESH: Dermatitis, Exfoliative / genetics, Skin Diseases, Genetic, MESH: Epidermis / pathology, MESH: Skin Diseases, Genetic / genetics, Up-Regulation, 3. Good health, MESH: Glycoproteins / genetics, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Intercellular Signaling Peptides and Proteins, Female, Dermatitis, Exfoliative, Down-Regulation, Dermatology, Computational biology, MESH: Transcriptome / genetics, Biology, 03 medical and health sciences, MESH: Gene Expression Profiling, Text mining, Downregulation and upregulation, Animals, Humans, MESH: Intercellular Signaling Peptides and Proteins, Molecular Biology, Gene, MESH: Mice, Glycoproteins, MESH: Humans, business.industry, Gene Expression Profiling, Cell Biology, Fold change, Gene expression profiling, Disease Models, Animal, 030104 developmental biology, Epidermis, MESH: Disease Models, Animal, business, MESH: Female
Abstract: International audience
Published: 2018

46. Severity of arterial defects in the retina correlates with the burden of intracerebral haemorrhage in COL4A1-related stroke

Author: Emmanuelle Plaisier, Nicolas Mezouar, Ambre Rochey, Anne Joutel, Julien Ratelade, Valérie Domenga-Denier, Génétique et Physiopathologie des Maladies Cérébro-Vasculaires (U1161 / UMR_S 1161), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Des Maladies Rénales Rares aux Maladies Fréquentes, Remodelage et Réparation, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), CHU Tenon [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), DHU NeuroVasc Sorbonne Paris-Cité, Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), and Dupuis, Christine
Subjects: 0301 basic medicine, MESH: Cerebral Hemorrhage, Pathology, Time Factors, [SDV]Life Sciences [q-bio], Penetrance, Degeneration (medical), MESH: Mice, Knockout, Muscle, Smooth, Vascular, MESH: Blood-Brain Barrier, chemistry.chemical_compound, 0302 clinical medicine, MESH: Penetrance, MESH: Animals, MESH: Endothelial Cells, MESH: Peptide Fragments, Stroke, Mice, Knockout, MESH: Genetic Predisposition to Disease, intracerebral haemorrhage, MESH: Myocytes, Smooth Muscle, MESH: Muscle, Smooth, Vascular, 3. Good health, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, Blood-Brain Barrier, retinal imaging, Disease Progression, MESH: Disease Progression, Collagen Type IV, medicine.medical_specialty, Mice, 129 Strain, Retinal Artery, COL4A1, Myocytes, Smooth Muscle, COL4A2, MESH: Stroke, Pathology and Forensic Medicine, 03 medical and health sciences, MESH: Mice, 129 Strain, MESH: Mice, Inbred C57BL, MESH: Cell Proliferation, Parenchyma, medicine, Animals, Genetic Predisposition to Disease, Pathological, MESH: Collagen Type IV, smooth muscle cell, Cell Proliferation, Cerebral Hemorrhage, Basement membrane, Retina, MESH: Retinal Artery, business.industry, MESH: Time Factors, Endothelial Cells, Retinal, medicine.disease, basement membrane, Peptide Fragments, Mice, Inbred C57BL, Disease Models, Animal, 030104 developmental biology, chemistry, MESH: Disease Models, Animal, business, 030217 neurology & neurosurgery
Abstract: International audience; Mutations in the α1 (COL4A1) or α2 (COL4A2) chains of collagen type IV, a major component of the vascular basement membrane, cause intracerebral haemorrhages with variable expressivity and reduced penetrance by mechanisms that remain poorly understood. Here we sought to investigate the cellular mechanisms of COL4A1-related intracerebral haemorrhage and identify a marker for haemorrhage risk stratification. A combination of histological, immunohistochemical, and electron microscopy analyses were used to analyse the brain parenchyma, cerebrovasculature, and retinal vessels of mice expressing the disease-causing COL4A1 p.G498V mutation. Mutant mice developed cerebral microhaemorrhages and macroscopic haemorrhages (macrohaemorrhages), the latter with reduced penetrance, mimicking the human disease. Microhaemorrhages that occurred in early postnatal life were associated with a transient, generalized increase in blood-brain barrier permeability at the level of capillaries. Macrohaemorrhages, which occurred later in life, originated from deep brain arteries with focal loss of smooth muscle cells. Similar smooth muscle cell loss was detected in retinal arteries, and a time-course analysis of arterial lesions showed that smooth muscle cells are recruited normally in arterial wall during development, but undergo progressive apoptosis-mediated degeneration. By assessing in parallel the extent of these retinal arterial lesions and the presence/absence of macrohaemorrhages, we found that the arterial lesion load in the retina is strongly correlated with the burden of macrohaemorrhages. We conclude that microhaemorrhages and macrohaemorrhages are driven by two distinct mechanisms. Moreover, smooth muscle cell degeneration is a critical factor underlying the partial penetrance of COL4A1-related macrohaemorrhages, and retinal imaging is a promising tool for identifying high-risk patients. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Published: 2018
Full Text: View/download PDF

47. LipL21 lipoprotein binding to peptidoglycan enables Leptospira interrogans to escape NOD1 and NOD2 recognition

Author: Ratet, Gwenn, Santecchia, Ignacio, Fanton d'Andon, Martine, Vernel-Pauillac, Frédérique, Wheeler, Richard, Lenormand, Pascal, Fischer, Frédéric, Lechat, Pierre, Haake, David, Picardeau, Mathieu, Boneca, Ivo, Werts, Catherine, Biologie et Génétique de la Paroi bactérienne - Biology and Genetics of Bacterial Cell Wall (BGPB), Institut Pasteur [Paris] (IP), Protéomique (Plate-Forme), Pathogenèse de Helicobacter, Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), David Geffen School of Medicine [Los Angeles], University of California [Los Angeles] (UCLA), University of California (UC)-University of California (UC), Biologie des Spirochètes / Biology of Spirochetes, This work was supported by Institut Pasteur and an ERC starting grant (PGNfromSHAPEtoVIR 202283) to IGB. GR’s PhD salary was from the DIM Malinf Ile de France to CW. IS was supported by a stipend from the Pasteur - Paris University (PPU) International PhD Program and Institut Carnot., European Project: 202283,EC:FP7:ERC,ERC-2007-StG,PGNFROMSHAPETOVIR(2008), Coburn, Jenifer, Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and University of California-University of California
Subjects: Male, MESH: Leptospira interrogans, Polymers, Nod2 Signaling Adaptor Protein, Pathology and Laboratory Medicine, MESH: Leptospira, Biochemistry, Transgenic, Mass Spectrometry, Mice, MESH: Nod2 Signaling Adaptor Protein, MESH: Animals, MESH: Immune Evasion, Aetiology, lcsh:QH301-705.5, Leptospira, Bacterial, MESH: Leptospirosis, Bacterial Pathogens, Medical Microbiology, Physical Sciences, Bacterial Outer Membrane Proteins, Materials by Structure, 1.1 Normal biological development and functioning, Knockout, Lipoproteins, Materials Science, Immunology, Peptidoglycan, Microbiology, Species Specificity, MESH: Protein Binding, MESH: Species Specificity, Humans, Leptospirosis, Microbial Pathogens, Immune Evasion, MESH: Mass Spectrometry, MESH: Humans, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Bacteria, Inflammatory and immune system, MESH: Bacterial Outer Membrane Proteins, Organisms, Immunity, Biology and Life Sciences, Proteins, Peptidoglycans, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Immunity, Innate, Mice, Inbred C57BL, Mutation, Leptospira interrogans, lcsh:RC581-607, MESH: Female, MESH: Signal Transduction, Physiology, Complement System, MESH: Nod1 Signaling Adaptor Protein, MESH: Virulence, Inbred C57BL, MESH: Mice, Knockout, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Immune Receptors, Immune Physiology, Nod1 Signaling Adaptor Protein, Medicine and Health Sciences, 2.1 Biological and endogenous factors, Innate, Mice, Knockout, Liquid Chromatography, Immune System Proteins, Virulence, MESH: Peptidoglycan, Chromatographic Techniques, MESH: Lipoproteins, Leptospira Interrogans, Chemistry, Macromolecules, MESH: Immunity, Innate, Female, Pathogens, Research Article, Signal Transduction, Protein Binding, lcsh:Immunologic diseases. Allergy, MESH: Mutation, MESH: Mice, Transgenic, Mice, Transgenic, Research and Analysis Methods, Underpinning research, MESH: Mice, Inbred C57BL, Virology, Animals, Antigens, MESH: Mice, Antigens, Bacterial, Cell Biology, Polymer Chemistry, MESH: Male, High Performance Liquid Chromatography, lcsh:Biology (General), Immune System, Pattern Recognition Receptors, MESH: Antigens, Bacterial
Abstract: Leptospirosis is a widespread zoonosis, potentially severe in humans, caused by spirochetal bacteria, Leptospira interrogans (L. interrogans). Host defense mechanisms involved in leptospirosis are poorly understood. Recognition of lipopolysaccharide (LPS) and lipoproteins by Toll-Like Receptors (TLR)4 and TLR2 is crucial for clearance of leptospires in mice, yet the role of Nucleotide Oligomerization Domain (NOD)-like receptors (NOD)1 and NOD2, recognizing peptidoglycan (PG) fragments has not previously been examined. Here, we show that pathogenic leptospires escape from NOD1 and NOD2 recognition both in vitro and in vivo, in mice. We found that leptospiral PG is resistant to digestion by certain hydrolases and that a conserved outer membrane lipoprotein of unknown function, LipL21, specific for pathogenic leptospires, is tightly bound to the PG. Leptospiral PG prepared from a mutant not expressing LipL21 (lipl21-) was more readily digested than the parental or complemented strains. Muropeptides released from the PG of the lipl21- mutant, or prepared using a procedure to eliminate the LipL21 protein from the PG of the parental strain, were recognized in vitro by the human NOD1 (hNOD1) and NOD2 (hNOD2) receptors, suggesting that LipL21 protects PG from degradation into muropeptides. LipL21 expressed in E. coli also resulted in impaired PG digestion and NOD signaling. We found that murine NOD1 (mNOD1) did not recognize PG of L. interrogans. This result was confirmed by mass spectrometry showing that leptospiral PG was primarily composed of MurTriDAP, the natural agonist of hNOD1, and contained only trace amounts of the tetra muropeptide, the mNOD1 agonist. Finally, in transgenic mice expressing human NOD1 and deficient for the murine NOD1, we showed enhanced clearance of a lipl21- mutant compared to the complemented strain, or to what was observed in NOD1KO mice, suggesting that LipL21 facilitates escape from immune surveillance in humans. These novel mechanisms allowing L. interrogans to escape recognition by the NOD receptors may be important in circumventing innate host responses., Author summary Leptospirosis is a widespread zoonosis caused by spirochetal bacteria, Leptospira interrogans (L. interrogans). L. interrogans are primarily extracellular pathogens although some reports suggest they may replicate within macrophages. In humans, leptospirosis can cause mild or severe disease, potentially leading to death, although rats or mice, which constitute the reservoir, are asymptomatic carriers. Host defense mechanisms involved in leptospirosis remain poorly understood. Toll-Like Receptor (TLR)2 and TLR4 are crucial for the clearance of L. interrogans, but the role of the cytosolic NOD receptors in leptospirosis is unknown. Here, we report that pathogenic leptospires escape the sensing of bacterial peptidoglycan through the NOD response. We found that an outer membrane lipoprotein of L. interrogans binds to and protects the peptidoglycan from degradation into muropeptides, thereby blocking signaling through NOD proteins. Moreover, in absence of this lipoprotein, the peptidoglycan of L. interrogans is properly sensed by human NOD1 but not by murine NOD1. This is due to the near absence of muramyl tetrapeptide, the murine NOD1 agonist, in the peptidoglycan of pathogenic leptospires. These novel mechanisms of NOD avoidance may facilitate the escape of leptospires from the innate immune system of their hosts.
Published: 2017

48. A human immune system mouse model with robust lymph node development

Author: Hugo Mouquet, Grégory Jouvion, Helene Strick-Marchand, Ai Ing Lim, Nicolas Serafini, Zacarias Garcia, Philippe Bousso, Yan Li, Daniela Finke, Olivier Schwartz, Timothée Bruel, Ayrin Kök, Franck Bourgade, Guillemette Masse-Ranson, Thierry Hieu, Antoine Toubert, Mathilde Dusséaux, James P. Di Santo, Immunité Innée - Innate Immunity, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Dynamiques des Réponses immunes - Dynamics of Immune Responses, Virus et Immunité - Virus and immunity, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Vaccine Research Institute (VRI), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Réponse humorale aux pathogènes, Neuropathologie expérimentale / Experimental neuropathology, Institut Pasteur [Paris]-Université de Paris (UP), Animalerie centrale (Plate-forme), Institut Pasteur [Paris], Service d'Immunologie et d'Histocompatibilité, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université de Paris (UP), Developmental Immunology, University of Basel (Unibas), This work was supported by the Agence Nationale de la Recherche (ANR) programme RPIB (Im_HIS to J.P.D.), the Laboratoire d’Excellence REVIVE (grant ANR-10-LABX-73 to Y.L. and J.P.D.), the Laboratoire d’Excellence Vaccine Research Institute (grant ANR-10-LABX-77 to T.B. and O.S.), the Laboratoire d’Excellence Milieu Intérieur (grant ANR-10-LABX-69 to H.M.), the Laboratoire d’Excellence IBEID (grant ANR-10-LABX-62 to O.S.), the Agence Nationale de Recherche sur le SIDA et les hepatitis (grant 15465 to O.S.), the European Commission Seventh Framework Programme no. 305578 (PathCO, to J.P.D.), the European Research Council (grant ERC-2013-StG 337146 to A.K. and T.H., grant ERC-2017-AvG 771167 to P.B.), Gilead Sciences (grant 00397 to G.M.-R. and J.P.D.), the Institut Pasteur (Core grant to J.P.D., G5 Program to H.M.), and INSERM (Core grant to J.P.D.)., ANR-10-LABX-0073,REVIVE,Stem Cells in Regenerative Biology and Medicine(2010), ANR-10-LABX-0077,VRI,Initiative for the creation of a Vaccine Research Institute(2010), ANR-10-LABX-0069,MILIEU INTERIEUR,GENETIC & ENVIRONMENTAL CONTROL OF IMMUNE PHENOTYPE VARIANCE: ESTABLISHING A PATH TOWARDS PERSONALIZED MEDICINE(2010), ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), European Project: 305578,EC:FP7:HEALTH,FP7-HEALTH-2012-INNOVATION-1,PATHCO(2012), European Project: 337146,EC:FP7:ERC,ERC-2013-StG,HUMANTIVIRUSES(2014), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Virus et Immunité - Virus and immunity (CNRS-UMR3569), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Vaccine Research Institute [Créteil, France] (VRI), Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Institut Pasteur [Paris] (IP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Cité (UPCité), and Institut Pasteur [Paris]-Université Paris Cité (UPCité)
Subjects: 0301 basic medicine, Male, T-Lymphocytes, MESH: Lymph Nodes, HIV Infections, Biochemistry, MESH: Mice, Knockout, MESH: HIV-1, Mice, MESH: Animals, Lymph node, Mice, Knockout, MESH: Cytokines, B-Lymphocytes, Mice, Inbred BALB C, MESH: Virus Latency, MESH: HIV Infections, 3. Good health, Virus Latency, medicine.anatomical_structure, Lymphatic system, [SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology, MESH: Immunoglobulin Class Switching, Cytokines, Female, Lymph, Biotechnology, Interleukin Receptor Common gamma Subunit, MESH: Mice, Transgenic, Transgene, MESH: Mice, Inbred BALB C, Mice, Transgenic, Biology, MESH: Models, Immunological, 03 medical and health sciences, Immune system, Thymic Stromal Lymphopoietin, Immunity, MESH: Mice, Inbred C57BL, MESH: B-Lymphocytes, medicine, Animals, Humans, Secondary lymphoid tissue, Molecular Biology, MESH: Mice, MESH: Humans, MESH: Interleukin Receptor Common gamma Subunit, Models, Immunological, Cell Biology, Immunoglobulin Class Switching, MESH: Male, Mice, Inbred C57BL, 030104 developmental biology, MESH: T-Lymphocytes, Immunoglobulin class switching, Immunology, HIV-1, Lymph Nodes, MESH: Female
Abstract: International audience; Lymph nodes (LNs) facilitate the cellular interactions that orchestrate immune responses. Human immune system (HIS) mice are powerful tools for interrogation of human immunity but lack secondary lymphoid tissue (SLT) as a result of a deficiency in Il2rg-dependent lymphoid tissue inducer cells. To restore LN development, we induced expression of thymic-stromal-cell-derived lymphopoietin (TSLP) in a Balb/c Rag2-/-Il2rg-/-SirpaNOD (BRGS) HIS mouse model. The resulting BRGST HIS mice developed a full array of LNs with compartmentalized human B and T cells. Compared with BRGS HIS mice, BRGST HIS mice have a larger thymus, more mature B cells, and abundant IL-21-producing follicular helper T (TFH) cells, and show enhanced antigen-specific responses. Using BRGST HIS mice, we demonstrated that LN TFH cells are targets of acute HIV infection and represent a reservoir for latent HIV. In summary, BRGST HIS mice reflect the effects of SLT development on human immune responses and provide a model for visualization and interrogation of regulators of immunity.
Published: 2017

49. Ecto-5′-Nucleotidase (CD73) Deficiency in Mycobacterium tuberculosis-Infected Mice Enhances Neutrophil Recruitment

Author: Grégory Jouvion, Laetitia Petit-Jentreau, Brigitte Gicquel, Patricia Charles, Laleh Majlessi, Ludovic Tailleux, Génétique mycobactérienne - Mycobacterial genetics, Institut Pasteur [Paris] (IP), Université Paris Diderot, Sorbonne Paris Cité, Paris, France, Université Paris Diderot - Paris 7 (UPD7), Histopathologie humaine et Modèles animaux, and Pathogénomique mycobactérienne intégrée
Subjects: MESH: Mycobacterium tuberculosis, Immunology, MESH: Flow Cytometry, Enzyme-Linked Immunosorbent Assay, Inflammation, MESH: Mice, Knockout, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, Microbiology, Proinflammatory cytokine, 5'-nucleotidase, Mycobacterium tuberculosis, Mice, Immune system, MESH: Mice, Inbred C57BL, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, medicine, Animals, MESH: Animals, MESH: Mice, 5'-Nucleotidase, Tuberculosis, Pulmonary, MESH: Tuberculosis, Pulmonary, Mice, Knockout, Host Response and Inflammation, biology, Purinergic receptor, MESH: Enzyme-Linked Immunosorbent Assay, Flow Cytometry, biology.organism_classification, Adenosine, Mice, Inbred C57BL, MESH: Neutrophil Infiltration, Disease Models, Animal, Infectious Diseases, Neutrophil Infiltration, Female, Parasitology, Tumor necrosis factor alpha, MESH: Disease Models, Animal, medicine.symptom, MESH: 5'-Nucleotidase, MESH: Female, medicine.drug
Abstract: The immune system needs safeguards that prevent collateral tissue damage mediated by the immune system while enabling an effective response against a pathogen. The purinergic pathway is one such mechanism and finely modulates inflammation by sensing nucleotides in the environment. Extracellular ATP is considered to be a danger signal leading to a proinflammatory response, whereas adenosine is immunosuppressive. CD73, also called ecto-5′-nucleotidase, occupies a strategic position in this pathway, as it is the main enzyme responsible for the generation of adenosine from ATP. Here, we explore the role of CD73 during tuberculosis, a disease characterized by an immune response that is harmful to the host and unable to eradicate Mycobacterium tuberculosis . Using CD73 knockout (KO) mice, we found that CD73 regulates the response to M. tuberculosis infection in vitro and in vivo . Mycobacterium-infected murine macrophages derived from CD73 KO mice secrete more keratinocyte chemoattractant (KC), tumor necrosis factor alpha (TNF-α), and interleukin-6 (IL-6) and release less vascular endothelial growth factor (VEGF) upon ATP stimulation than do those derived from wild-type (WT) mice. In vivo , CD73 limits the early influx of neutrophils to the lungs without affecting bacterial growth and dissemination. Collectively, our results support the view that CD73 fine-tunes antimycobacterial immune responses.
Published: 2015

50. Tissue kallikrein is required for the cardioprotective effect of Cyclosporin A in myocardial ischemia in the mouse

Author: C. Richer, M.-P. Vincent, G. Youcef, François Alhenc-Gelas, Anne Pizard, Elise Belaidi, M. Clemessy, L. Fazal, Ludovic Waeckel, G. Zadigue, Michel Ovize, Centre de Recherche des Cordeliers (CRC), Université Pierre et Marie Curie - Paris 6 (UPMC)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Cardiovasculaire, métabolisme, diabétologie et nutrition (CarMeN), Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National de la Recherche Agronomique (INRA), Défaillance Cardiovasculaire Aiguë et Chronique (DCAC), Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lorraine (UL), Centre d'investigation clinique [Nancy] (CIC), Université Paris Diderot - Paris 7 (UPD7)-École pratique des hautes études (EPHE)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM), CIC-Nancy, Institut Lorrain du Coeur et des Vaisseaux Louis Mathieu [Nancy]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Pierre et Marie Curie - Paris 6 (UPMC)-École Pratique des Hautes Études (EPHE), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hospices Civils de Lyon (HCL), and Université de Lorraine (UL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy)
Subjects: Male, MESH: Signal Transduction, Myocardial Ischemia, Gene Expression, MESH: Myocytes, Cardiac, Mitochondrion, Pharmacology, MESH: Voltage-Dependent Anion Channel 1, Mitochondrial Membrane Transport Proteins, MESH: Mice, Knockout, MESH: Cyclosporine, Biochemistry, Mitochondria, Heart, Oxidative Phosphorylation, Cyclophilins, Mice, chemistry.chemical_compound, Cyclosporin a, Myocytes, Cardiac, MESH: Animals, Ischemic Postconditioning, Inner mitochondrial membrane, MESH: Heterozygote, Mice, Knockout, Cardioprotection, Tissue kallikrein, MPTP, Homozygote, MESH: Mitochondrial Membrane Transport Proteins, Mitochondria, Knockout mouse, Cyclosporine, cardiovascular system, Cardiac ischemia–reperfusion, MESH: Myocardial Ischemia, MESH: Mitochondria, Heart, Tissue Kallikreins, Cyclophilin D, Signal Transduction, MESH: Homozygote, Heterozygote, Cardiotonic Agents, MESH: Myocardium, MESH: Gene Expression, MESH: Rats, MESH: Tissue Kallikreins, Ischemia, Biology, MESH: Cyclophilins, MESH: Ischemic Postconditioning, [SDV.MHEP.CSC]Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, MESH: Oxidative Phosphorylation, medicine, Animals, cardiovascular diseases, MESH: Mice, Mitochondrial Permeability Transition Pore, Myocardium, Voltage-Dependent Anion Channel 1, MESH: Cardiotonic Agents, medicine.disease, MESH: Male, Rats, Cyclosporin A, Mitochondrial permeability transition pore, chemistry
Abstract: International audience; Clinical and experimental studies suggest that pharmacological postconditioning with Cyclosporin A (CsA) reduces infarct size in cardiac ischemia and reperfusion. CsA interacts with Cyclophilin D (CypD) preventing opening of the mitochondrial permeability transition pore (mPTP). Tissue kallikrein (TK) and its products kinins are involved in cardioprotection in ischemia. CypD knockout mice are resistant to the cardioprotective effects of both CsA and kinins suggesting common mechanisms of action. Using TK gene knockout mice, we investigated whether the kallikrein-kinin system is involved in the cardioprotective effect of CsA. Homozygote and heterozygote TK deficient mice (TK(-/-), TK(+/-)) and wild type littermates (TK(+/+)) were subjected to cardiac ischemia-reperfusion with and without CsA postconditioning. CsA reduced infarct size in TK(+/+) mice but had no effect in TK(+/-) and TK(-/-) mice. Cardiac mitochondria isolated from TK(-/-) mice had indistinguishable basal oxidative phosphorylation and calcium retention capacity compared to TK(+/+) mice but were resistant to CsA inhibition of mPTP opening. TK activity was documented in mouse heart and rat cardiomyoblasts mitochondria. By proximity ligation assay TK was found in close proximity to the mitochondrial membrane proteins VDAC and Tom22, and CypD. Thus, partial or total deficiency in TK induces resistance to the infarct size reducing effect of CsA in cardiac ischemia in mice, suggesting that TK level is a critical factor for cardioprotection by CsA. TK is required for the mitochondrial action of CsA and may interact with CypD. Genetic variability in TK activity has been documented in man and may influence the cardioprotective effect of CsA.
Published: 2015

Catalog

Books, media, physical & digital resources

See catalog results

Searchworks

Select search scope, currently: Articles Catalog books, media & more in Jio Institute collections Articles journal articles & other e-resources

Search

Search Constraints

Refine your results

Search Limiters

Topic

Publication Year Range

Language

Journal

Database

Publisher

908 results on '"MESH: Mice, Knockout"'

Search Results

Catalog

Select search scope, currently: Articles

Catalog

books, media & more in Jio Institute collections

Articles

journal articles & other e-resources