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1. Mice Humanized for MHC and hACE2 with High Permissiveness to SARS-CoV-2 Omicron Replication

Author: Fabien Le Chevalier, Pierre Authié, Sébastien Chardenoux, Maryline Bourgine, Benjamin Vesin, Delphine Cussigh, Yohann Sassier, Ingrid Fert, Amandine Noirat, Kirill Nemirov, François Anna, Marion Bérard, Françoise Guinet, David Hardy, Pierre Charneau, François Lemonnier, Francina Langa-Vives, Laleh Majlessi, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Animalerie Centrale (plateforme) - Central Animal Facility (platform) (C2RA), Lymphocytes et Immunité - Lymphocytes and Immunity, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Plate-Forme d’Histopathologie / Histopathology Platform, Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), and Institut Pasteur et Theravectys
Subjects: HLA, Infectious Diseases, SARS-CoV-2, [SDV]Life Sciences [q-bio], Immunology, Transgenesis, Omicron variant, Microbiology, COVID-19 animal model
Abstract: International audience; Human Angiotensin-Converting Enzyme 2 (hACE2) is the major receptor enabling host cell invasion by SARS-CoV-2 via interaction with Spike. The murine ACE2 does not interact efficiently with SARS CoV-2 Spike and therefore the laboratory mouse strains are not permissive to SARS-CoV-2 replication. Here, we generated new hACE2 transgenic mice, which harbor the hACE2 gene under the human keratin 18 promoter, in “HHD-DR1” background. HHD-DR1 mice are fully devoid of murine Major Histocompatibility Complex (MHC) molecules of class-I and -II and express only MHC molecules from Human Leukocyte Antigen (HLA) HLA 02.01, DRA01.01, DRB1.01.01 alleles, widely expressed in human populations. We selected three transgenic strains, with various hACE2 mRNA expression levels and distinctive profiles of lung and/or brain permissiveness to SARS-CoV-2 replication. These new hACE2 transgenic strains display high permissiveness to the replication of SARS-CoV-2 Omicron sub-variants, while the previously available B6.K18-ACE22Prlmn/JAX mice have been reported to be poorly susceptible to infection with Omicron. As a first application, one of these MHC- and ACE2-humanized strains was successfully used to show the efficacy of a lentiviral-based COVID-19 vaccine
Published: 2023
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2. Clonal hematopoiesis driven by chromosome 1q/MDM4 trisomy defines a canonical route toward leukemia in Fanconi anemia

Author: Marie Sebert, Stéphanie Gachet, Thierry Leblanc, Alix Rousseau, Olivier Bluteau, Rathana Kim, Raouf Ben Abdelali, Flore Sicre de Fontbrune, Loïc Maillard, Carèle Fedronie, Valentine Murigneux, Léa Bellenger, Naira Naouar, Samuel Quentin, Lucie Hernandez, Nadia Vasquez, Mélanie Da Costa, Pedro H. Prata, Lise Larcher, Marie de Tersant, Matthieu Duchmann, Anna Raimbault, Franck Trimoreau, Odile Fenneteau, Wendy Cuccuini, Nathalie Gachard, Nathalie Auger, Giulia Tueur, Maud Blanluet, Claude Gazin, Michèle Souyri, Francina Langa Vives, Aaron Mendez-Bermudez, Hélène Lapillonne, Etienne Lengline, Emmanuel Raffoux, Pierre Fenaux, Lionel Adès, Edouard Forcade, Charlotte Jubert, Carine Domenech, Marion Strullu, Bénédicte Bruno, Nimrod Buchbinder, Caroline Thomas, Arnaud Petit, Guy Leverger, Gérard Michel, Marina Cavazzana, Eliane Gluckman, Yves Bertrand, Nicolas Boissel, André Baruchel, Jean-Hugues Dalle, Emmanuelle Clappier, Eric Gilson, Ludovic Deriano, Sylvie Chevret, François Sigaux, Gérard Socié, Dominique Stoppa-Lyonnet, Hugues de Thé, Christophe Antoniewski, Dominique Bluteau, Régis Peffault de Latour, Jean Soulier, Institut de Recherche Saint-Louis - Hématologie Immunologie Oncologie (Département de recherche de l’UFR de médecine, ex- Institut Universitaire Hématologie-IUH) (IRSL), Université Paris Cité (UPCité), Hopital Saint-Louis [AP-HP] (AP-HP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Génomes, biologie cellulaire et thérapeutiques (GenCellDi (U944 / UMR7212)), Collège de France (CdF (institution))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), AP-HP Hôpital universitaire Robert-Debré [Paris], Recherche clinique appliquée à l'hématologie (URP_3518), Intégrité du génome, immunité et cancer - Genome integrity, Immunity and Cancer, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), CHU Limoges, Institut Gustave Roussy (IGR), Unité de génétique et biologie des cancers (U830), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de Recherche en Génomique Humaine (CNRGH), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Hématopoïèse normale et pathologique : émergence, environnement et recherche translationnelle [Paris] ((UMR_S1131 / U1131)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), Institut de Recherche sur le Cancer et le Vieillissement (IRCAN), 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)-Centre National de la Recherche Scientifique (CNRS)-Université Côte d'Azur (UCA), CHU Nice [Cimiez], Hôpital Cimiez [Nice] (CHU), CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Hôpitaux universitaires Est parisien [AP-HP], CHU Bordeaux [Bordeaux], Hospices Civils de Lyon (HCL), 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), Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), CHU Rouen, Normandie Université (NU), Centre hospitalier universitaire de Nantes (CHU Nantes), Centre d'études et de recherche sur les services de santé et la qualité de vie (CEReSS), Aix Marseille Université (AMU), Hôpital de la Timone [CHU - APHM] (TIMONE), CIC NECKER BT (CIC 1416), Immunologie humaine, physiopathologie & immunothérapie (HIPI (UMR_S_976 / U976)), Collège de France (CdF (institution)), École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL), This study received support from the European Research Council (ERC) Consolidator Grant to J.S. (CEVAL-311660), the FP7 Eurofancolen program (HEALTH-F5-2012-305421), the ANR program PACRI (Projet alliance parisienne des instituts de recherche en cancérologie), the CONECT-AML (Collaborative Network for Children and Teenagers with Acute Myeloid Leukemia) program supported by a grant from the Institut National du Cancer (INCa), Fondation ARC, Ligue nationale contre le cancer, and Laurette Fugain (INCa-ARC-LIGUE_11905) to J.S. and C.A., and the Association Française pour la Maladie de Fanconi (AFMF) grants 'Histoire naturelle de la maladie de Fanconi' to R.P.L. and J.S., 'Modélisation de la transformation leucémique dans la maladie de Fanconi' to D.B., and 'Cribles fonctionnels à haut débit de gènes modificateurs de la maladie de Fanconi' to C.G. M.S. was supported by the AVIESAN-INCa Program 'Formation à la Recherche Translationnelle,' and A.R. by a grant from the Fondation ARC. The work in E.G.’s lab is supported by Fondation ARC and ANR Telochrom. The work in L.D.’s lab is supported by INCa (PLBIO16-181) and ERC (310917)., ANR-11-PHUC-0002,PACRI,Alliance Parisienne des Instituts de Recherche en Cancérologie(2011), European Project: 311660,EC:FP7:ERC,ERC-2012-StG_20111109,CEVAL(2013), and European Project: 305421,EC:FP7:HEALTH,FP7-HEALTH-2012-INNOVATION-1,EUROFANCOLEN(2013)
Subjects: MDM4, Fanconi anemia, precision medicine, [SDV]Life Sciences [q-bio], Genetics, leukemia, clonal hematopoiesis, Molecular Medicine, Cell Biology, TP53, mutational signature, genomic instability, BRCA2
Abstract: International audience; Fanconi anemia (FA) patients experience chromosome instability, yielding hematopoietic stem/progenitor cell (HSPC) exhaustion and predisposition to poor-prognosis myeloid leukemia. Based on a longitudinal cohort of 335 patients, we performed clinical, genomic, and functional studies in 62 patients with clonal evolution. We found a unique pattern of somatic structural variants and mutations that shares features of BRCA-related cancers, the FA-hallmark being unbalanced, microhomology-mediated translocations driving copy-number alterations. Half the patients developed chromosome 1q gain, driving clonal hematopoiesis through MDM4 trisomy downmodulating p53 signaling later followed by secondary acute myeloid lukemia genomic alterations. Functionally, MDM4 triplication conferred greater fitness to murine and human primary FA HSPCs, rescued inflammation-mediated bone marrow failure, and drove clonal dominance in FA mouse models, while targeting MDM4 impaired leukemia cells in vitro and in vivo. Our results identify a linear route toward secondary leukemogenesis whereby early MDM4-driven downregulation of basal p53 activation plays a pivotal role, opening monitoring and therapeutic prospects.
Published: 2023
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3. SPICE‐Met : profiling and imaging energy metabolism at the single‐cell level using a fluorescent reporter mouse

Author: Erica Russo, Fabrice Lemaître, Béatrice Corre, Aleksandra S Chikina, Francina Langa‐Vives, Philippe Bousso, Dynamiques des Réponses immunes - Dynamics of Immune Responses, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Vaccine Research Institute [Créteil, France] (VRI), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), The work was supported by Institut Pasteur, INSERM, the Vaccine Research Institute in Créteil, France, and an advanced grant (ENLIGHTEN) from the European Research Council. E.R. was supported by fellowships from the Swiss National Science Foundation and the Vaccine Research Institute., We thank members of the Bousso laboratory for the critical review of the manuscript. We thank CB_UTechS at Institut Pasteur for support in conducting the present study., Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Vaccine Research Institute (VRI), and Institut Pasteur [Paris]-Université Paris Cité (UPCité)
Subjects: General Immunology and Microbiology, General Neuroscience, immunometabolism, imaging, Mice, Transgenic, glycolysis, OXPHOS, Oxidative Phosphorylation, General Biochemistry, Genetics and Molecular Biology, Adenosine Diphosphate, Mice, Adenosine Triphosphate, Animals, [SDV.IMM]Life Sciences [q-bio]/Immunology, Energy Metabolism, Molecular Biology, energy
Abstract: International audience; The regulation of cellular energy metabolism is central to most physiological and pathophysiological processes. However, most current methods have limited ability to functionally probe metabolic pathways in individual cells. Here, we describe SPICE-Met (Single-cell Profiling and Imaging of Cell Energy Metabolism), a method for profiling energy metabolism in single cells using flow cytometry or imaging. We generated a transgenic mouse expressing PercevalHR, a fluorescent reporter for cellular ATP:ADP ratio. Modulation of PercevalHR fluorescence with metabolic inhibitors was used to infer the dependence of energy metabolism on oxidative phosphorylation and glycolysis in defined cell populations identified by flow cytometry. We applied SPICE-Met to analyze T-cell memory development during vaccination. Finally, we used SPICE-Met in combination with real-time imaging to dissect the heterogeneity and plasticity of energy metabolism in single macrophages ex vivo and identify three distinct metabolic patterns. Functional probing of energy metabolism with single-cell resolution should greatly facilitate the study of immunometabolism at a steady state, during disease pathogenesis or in response to therapy.
Published: 2022
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4. Rab35-regulated lipid turnover by myotubularins represses mTORC1 activity and controls myelin growth

Author: Kerstin Klinkert, Volker Haucke, Alessandra Bolino, Genaro Patino-Lopez, Linda Sawade, Francina Langa-Vives, Federica Grandi, Marianna Mignanelli, Roberta Di Guardo, Arnaud Echard, Leibniz Forschungsinstitut für Molekulare Pharmakolgie = Leibniz Institute for Molecular Pharmacology [Berlin, Allemagne] (FMP), Leibniz Association, IRCCS Ospedale San Raffaele [Milan, Italy], Universita Vita Salute San Raffaele = Vita-Salute San Raffaele University [Milan, Italie] (UniSR), Hospital Infantil de México Federico Gómez (HIMFG), Trafic membranaire et Division cellulaire - Membrane Traffic and Cell Division, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Université (SU), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), Freie Universität Berlin, Charité - UniversitätsMedizin = Charité - University Hospital [Berlin], Supported by grants from the German Research Foundation (DFG) (SFB958/A01 and HA2686/15-1 to V.H.), from Muscular Dystrophy Association (MDA574294) and AFM-Telethon France (#21528) to A.B., V.H. and A.B. are funded by E-Rare-3 JTC 2017 and are members of the eRARE Consortium 'Treat-MTMs'. Part of this work has been supported by Institut Pasteur and CNRS to A.E., K.K. was supported by the Pasteur - Paris University (PPU) International PhD Program and by and a fellowship from FRM (FDT20150532389)., Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris]
Subjects: 0301 basic medicine, Myelin biology and repair, General Physics and Astronomy, GTPase, mTORC1, [SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC], Small GTPases, chemistry.chemical_compound, Myelin, 0302 clinical medicine, Charcot-Marie-Tooth Disease, Gene Knock-In Techniques, Lipid signalling, lcsh:Science, Myelin Sheath, Multidisciplinary, Hyperactivation, Chemistry, Protein Tyrosine Phosphatases, Non-Receptor, Phenotype, 3. Good health, Cell biology, medicine.anatomical_structure, Signal Transduction, congenital, hereditary, and neonatal diseases and abnormalities, Science, Primary Cell Culture, Down-Regulation, Mice, Transgenic, Mechanistic Target of Rapamycin Complex 1, General Biochemistry, Genetics and Molecular Biology, Article, 03 medical and health sciences, medicine, Animals, Humans, Phosphatidylinositol, Gene, General Chemistry, Nutrient signalling, Lipid Metabolism, nervous system diseases, 030104 developmental biology, HEK293 Cells, rab GTP-Binding Proteins, Astrocytes, Mutation, lcsh:Q, 030217 neurology & neurosurgery, Biogenesis, HeLa Cells
Abstract: Inherited peripheral neuropathies (IPNs) represent a broad group of disorders including Charcot-Marie-Tooth (CMT) neuropathies characterized by defects primarily arising in myelin, axons, or both. The molecular mechanisms by which mutations in nearly 100 identified IPN/CMT genes lead to neuropathies are poorly understood. Here we show that the Ras-related GTPase Rab35 controls myelin growth via complex formation with the myotubularin-related phosphatidylinositol (PI) 3-phosphatases MTMR13 and MTMR2, encoded by genes responsible for CMT-types 4B2 and B1 in humans, and found that it downregulates lipid-mediated mTORC1 activation, a pathway known to crucially regulate myelin biogenesis. Targeted disruption of Rab35 leads to hyperactivation of mTORC1 signaling caused by elevated levels of PI 3-phosphates and to focal hypermyelination in vivo. Pharmacological inhibition of phosphatidylinositol 3,5-bisphosphate synthesis or mTORC1 signaling ameliorates this phenotype. These findings reveal a crucial role for Rab35-regulated lipid turnover by myotubularins to repress mTORC1 activity and to control myelin growth., Charcot-Marie-Tooth (CMT) is an inherited peripheral neuropathy. Here, the authors show that Rab35 forms a complex with genes implicated in CMT, MTMR13 and MTMR2, which regulates myelin growth by controlling mTORC1 signaling through lipid turnover.
Published: 2020
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5. An intranasal lentiviral booster reinforces the waning mRNA vaccine-induced SARS-CoV-2 immunity that it targets to lung mucosa

Author: Benjamin Vesin, Jodie Lopez, Amandine Noirat, Pierre Authié, Ingrid Fert, Fabien Le Chevalier, Fanny Moncoq, Kirill Nemirov, Catherine Blanc, Cyril Planchais, Hugo Mouquet, Françoise Guinet, David Hardy, Francina Langa Vives, Christiane Gerke, François Anna, Maryline Bourgine, Laleh Majlessi, Pierre Charneau, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité), Immunologie humorale - Humoral Immunology, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Lymphocytes et Immunité - Lymphocytes and Immunity, Plate-Forme d’Histopathologie / Histopathology Platform, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Service développement de l'innovation - Innovation Office, Direction des Applications de la Recherche et des Relations industrielles - The Technology Transfer and Industrial Partnership Department (DARRI), Institut Pasteur [Paris] (IP)-Institut Pasteur [Paris] (IP), This work was supported by TheraVectys and Institut Pasteur., The authors are grateful to Dr. Marie José Quentin-Millet and Estelle Besson (TheraVectys) for discussion and advice, to Magali Tichit and Sabine Maurin for excellent technical assistance in preparing histological sections, to Sébastien Chardenoux (CIGM, Institut Pasteur) for excellent technical assistance in B6.K18-hACE2IP−THV mouse transgenesis, and to Marie Laure Loriquet and Dr. Paul-Henri Consigny (Institut Pasteur Medical Center) for their gift of the leftover Moderna vaccines, not unusable in humans, so that this study did not deprive any individual of a vaccine dose during the COVID-19 pandemic. The SARS-CoV2 variant Delta/2021/I7.2 200 was supplied by the Virus and Immunity Unit (Institut Pasteur, Paris, France) headed by Olivier Schwartz. The SARS-CoV-2 Omicron BA.1 variant was initially supplied by the Virus and Immunity Unit (Institut Pasteur, Paris, France) headed by Olivier Schwartz, and was provided to our laboratory by Matthieu Prot and Etienne Simon-Lorière (G5 Evolutionary Genomics of RNA Viruses, Institut Pasteur, Paris, France)., and Majlessi, Laleh
Subjects: COVID-19 Vaccines, lentiviral vaccine, Antibodies, Viral, Mice, [SDV.IMM.VAC] Life Sciences [q-bio]/Immunology/Vaccinology, Drug Discovery, Genetics, mucosal booster vaccine, Animals, Humans, SARS-CoV-2 emerging variants of concern, Lung, Molecular Biology, Pharmacology, Vaccines, Synthetic, Mucous Membrane, SARS-CoV-2, Vaccination, COVID-19, Viral Vaccines, Antibodies, Neutralizing, intranasal vaccination, Molecular Medicine, mucosal immunity, waning anti-COVID-19 immunity, mRNA Vaccines, [SDV.IMM.VAC]Life Sciences [q-bio]/Immunology/Vaccinology
Abstract: International audience; As the coronavirus disease 2019 (COVID-19) pandemic continues and new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern emerge, the adaptive immunity initially induced by the first-generation COVID-19 vaccines starts waning and needs to be strengthened and broadened in specificity. Vaccination by the nasal route induces mucosal, humoral, and cellular immunity at the entry point of SARS-CoV-2 into the host organism and has been shown to be the most effective for reducing viral transmission. The lentiviral vaccination vector (LV) is particularly suitable for this route of immunization owing to its non-cytopathic, non-replicative, and scarcely inflammatory properties. Here, to set up an optimized cross-protective intranasal booster against COVID-19, we generated an LV encoding stabilized spike of SARS-CoV-2 Beta variant (LV::SBeta-2P). mRNA vaccine-primed and -boosted mice, with waning primary humoral immunity at 4 months after vaccination, were boosted intranasally with LV::SBeta-2P. A strong boost effect was detected on cross-sero-neutralizing activity and systemic T cell immunity. In addition, mucosal anti-spike IgG and IgA, lung-resident B cells, and effector memory and resident T cells were efficiently induced, correlating with complete pulmonary protection against the SARS-CoV-2 Delta variant, demonstrating the suitability of the LV::SBeta-2P vaccine candidate as an intranasal booster against COVID-19. LV::SBeta-2P vaccination was also fully protective against Omicron infection of the lungs and central nervous system, in the highly susceptible B6.K18-hACE2IP-THV transgenic mice.
Published: 2022
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6. De novo germline mutation in the dual specificity phosphatase 10 gene accelerates autoimmune diabetes

Author: Fabrice Valette, Pere Santamaria, Jean-François Bach, Claudio Scazzocchio, Sophie Candon, Sebastien Lemoine, Sara Hildebrand, Peipei Hu, Bruce Beutler, Sylvaine You, Francina Langa-Vives, Stephen Aplin Lyon, Anne-Perrine Foray, Eva Marie Y. Moresco, Chun Hui Bu, Lucienne Chatenoud, Coralie Pecquet, Darui Xu, Cindy Marquet, Michael Dumas, Lindsay Scott, Tao Wang, Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), University of Texas Southwestern Medical Center [Dallas], Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), Biotechnologie et signalisation cellulaire (BSC), Université de Strasbourg (UNISTRA)-Institut de recherche de l'Ecole de biotechnologie de Strasbourg (IREBS)-Centre National de la Recherche Scientifique (CNRS), University of Calgary, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Imperial College London, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Funding was provided by NIH Grant R01 AI125581 (B.B.), NIH Grant U19 AI100627 (B.B.), European Research Council Advanced Grant, Hygiene No. 250290 (J.-F.B.), Institut National de la Santé et de la Recherche Scientifique (L.C.), Fondation Day Solvay (L.C.), and the Lyda Hill Foundation (B.B.)., European Project: 250290,EC:FP7:ERC,ERC-2009-AdG,HYGIENE(2010), LANGA VIVES, FRANCINA, and THE HYGIENE HYPOTHESIS : REVISITING THE CONCEPT BY INTEGRATING EPIDEMIOLOGY AND MECHANISTIC STUDIES - HYGIENE - - EC:FP7:ERC2010-08-01 - 2015-07-31 - 250290 - VALID
Subjects: Male, type 1 diabetes, Nod, Major histocompatibility complex, medicine.disease_cause, Autoimmune Diseases, Major Histocompatibility Complex, Islets of Langerhans, Mice, Germline mutation, Mice, Inbred NOD, medicine, Animals, Humans, Missense mutation, Genetic Predisposition to Disease, Germ-Line Mutation, NOD mice, Autoimmune disease, Genetics, Mutation, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, Multidisciplinary, biology, autoimmunity, Haplotype, Biological Sciences, medicine.disease, Mice, Inbred C57BL, Diabetes Mellitus, Type 1, Haplotypes, biology.protein, Dual-Specificity Phosphatases, Mitogen-Activated Protein Kinase Phosphatases, Female, genetic mapping, NOD mouse, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, Genome-Wide Association Study
Abstract: International audience; Significance: The vast majority of autoimmune diseases are polygenic, and causal loci uncovered by genetic-mapping studies explain only a minority of the heritable contribution to trait variation. Multiple explanations for this missing heritability include rare meaningful variants, rare copy number variations or deletions, epistasis, epigenetics, disease heterogeneity, and rare or infrequent variants that segregate within individual families (even within monozygotic twins). Here we demonstrate that experimental models of spontaneous autoimmune diseases may be invaluable tools to map rare germline variants impacting disease susceptibility traits. We identified a variant of the dual-specificity phosphatase 10 encoding gene that accelerates disease in an autoimmune type 1 diabetes model, the nonobese diabetic mouse.Abstract: Insulin-dependent or type 1 diabetes (T1D) is a polygenic autoimmune disease. In humans, more than 60 loci carrying common variants that confer disease susceptibility have been identified by genome-wide association studies, with a low individual risk contribution for most variants excepting those of the major histocompatibility complex (MHC) region (40 to 50% of risk); hence the importance of missing heritability due in part to rare variants. Nonobese diabetic (NOD) mice recapitulate major features of the human disease including genetic aspects with a key role for the MHC haplotype and a series of Idd loci. Here we mapped in NOD mice rare variants arising from genetic drift and significantly impacting disease risk. To that aim we established by selective breeding two sublines of NOD mice from our inbred NOD/Nck colony exhibiting a significant difference in T1D incidence. Whole-genome sequencing of high (H)- and low (L)-incidence sublines (NOD/NckH and NOD/NckL) revealed a limited number of subline-specific variants. Treating age of diabetes onset as a quantitative trait in automated meiotic mapping (AMM), enhanced susceptibility in NOD/NckH mice was unambiguously attributed to a recessive missense mutation of Dusp10, which encodes a dual specificity phosphatase. The causative effect of the mutation was verified by targeting Dusp10 with CRISPR-Cas9 in NOD/NckL mice, a manipulation that significantly increased disease incidence. The Dusp10 mutation resulted in islet cell down-regulation of type I interferon signature genes, which may exert protective effects against autoimmune aggression. De novo mutations akin to rare human susceptibility variants can alter the T1D phenotype.
Published: 2021
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7. Brain Cross‐Protection against SARS‐CoV‐2 Variants by a Lentiviral Vaccine in New Transgenic Mice

Author: Ku, Min-Wen, Authié, Pierre, Bourgine, Maryline, Anna, François, Noirat, Amandine, Moncoq, Fanny, Vesin, Benjamin, Nevo, Fabien, Lopez, Jodie, Souque, Philippe, Blanc, Catherine, Fert, Ingrid, Chardenoux, Sébastien, Lafosse, llta, Cussigh, Delphine, Hardy, David, Nemirov, Kirill, Guinet, Françoise, Vives, Francina Langa, Majlessi, Laleh, Charneau, Pierre, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris]-TheraVectys, Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], Neuropathologie expérimentale / Experimental neuropathology, Institut Pasteur [Paris]-Université de Paris (UP), Lymphocytes et Immunité - Lymphocytes and Immunity, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), This work was supported by the «URGENCE COVID-19» fundraising campaign of Institut Pasteur, TheraVectys and Agence Nationale de la Recherche (ANR) HuMoCID. M.W. Ku is part of the Pasteur - Paris University (PPU) International PhD Program and received funding from the Institut Carnot Pasteur Microbes & Santé, and the European Union's Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement No 665807., The authors are grateful to Pr S. van der Werf (National Reference Centre for Respiratory Viruses hosted by Institut Pasteur, Paris, France), F. Guivel-Benhassine and Pr O. Schwartz (Institut Pasteur) for providing the BetaCoV/France/IDF0372/2020 and Gamma (P.1) SARS-CoV-2 clinical isolates. The strain BetaCoV/France/IDF0372/2020 was supplied through the European Virus Archive goes Global (Evag) platform, a project that has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No 653316. The authors thank Pr G. Milon and Dr L.A. Chakrabarti for fruitful advice and discussion, Dr H. Mouquet and Dr C. Planchais for providing recombinant homotrimeric S proteins, Dr N. Escriou and Dr M. Gransagne for providing a plasmid containing the prefusion SCoV-2 sequence,M. Tichit and N. Dominique, for excellent technical assistance, respectively, in preparing histological sections and in animal immunization., ANR-20-COVI-0028,HuMoCID,Développement de modèles murins de COVID-19(2020), European Project: 665807,H2020,H2020-MSCA-COFUND-2014,PASTEURDOC(2015), Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité), Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Hardy, David, Développement de modèles murins de COVID-19 - - HuMoCID2020 - ANR-20-COVI-0028 - COVID-19 - VALID, and Institut Pasteur International Docotal Program - PASTEURDOC - - H20202015-10-01 - 2020-10-01 - 665807 - VALID
Subjects: Central Nervous System, Medicine (General), COVID-19 Vaccines, [SDV]Life Sciences [q-bio], Immunology, Mice, Transgenic, QH426-470, Antibodies, Viral, Article, Mice, R5-920, Genetics, Animals, Humans, SARS-CoV-2, Brain, COVID-19, Articles, hACE2 Transgenic Mice, Antibodies, Neutralizing, Olfactory Bulb, SARS-CoV-2 Emerging Variants of Concern, Microbiology, Virology & Host Pathogen Interaction, [SDV] Life Sciences [q-bio], Intranasal Vaccination, Spike Glycoprotein, Coronavirus, SARS‐CoV‐2 emerging variants of concern
Abstract: COVID‐19 vaccines already in use or in clinical development may have reduced efficacy against emerging SARS‐CoV‐2 variants. In addition, although the neurotropism of SARS‐CoV‐2 is well established, the vaccine strategies currently developed have not taken into account protection of the central nervous system. Here, we generated a transgenic mouse strain expressing the human angiotensin‐converting enzyme 2, and displaying unprecedented brain permissiveness to SARS‐CoV‐2 replication, in addition to high permissiveness levels in the lung. Using this stringent transgenic model, we demonstrated that a non‐integrative lentiviral vector, encoding for the spike glycoprotein of the ancestral SARS‐CoV‐2, used in intramuscular prime and intranasal boost elicits sterilizing protection of lung and brain against both the ancestral virus, and the Gamma (P.1) variant of concern, which carries multiple vaccine escape mutations. Beyond induction of strong neutralizing antibodies, the mechanism underlying this broad protection spectrum involves a robust protective T‐cell immunity, unaffected by the recent mutations accumulated in the emerging SARS‐CoV‐2 variants., A lentiviral vector‐based vaccine, used in prime and intranasal boost, induces strong B‐ and T‐cell immunity. This vaccine candidate protects against both pulmonary and neurological COVID‐19 caused by either ancestral or emerging SARS‐CoV‐2.
Published: 2021
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8. The antitumoral effects of chemerin are independent from leukocyte recruitment and mediated by inhibition of neoangiogenesis

Author: Diana Al Delbany, Anne Lefort, Virginie Robert, Marc Parmentier, Francina Langa, Maxime Vernimmen, Valérie Wittamer, Ingrid Dubois-Vedrenne, Olivier De Henau, Frédérick Libert, Institut de Recherche Interdisciplinaire en Biologie Humaine et moléculaire = Insitute of Interdisciplinary Research [Bruxelles, Belgique] (IRIBHM), Faculté de Médecine [Bruxelles] (ULB), Université libre de Bruxelles (ULB)-Université libre de Bruxelles (ULB), Walloon Excellence in Life sciences and BIOtechnology [Liège] (WELBIO), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), This work was supported by the Belgian programme on Interuniversity Poles of Attraction initiated by the Belgian State, Prime Minister’s Office, Science Policy Programming, the Fonds de la Recherche Scientifique Médicale of Belgium, Télévie, the Fondation Belge contre le Cancer and the Actions de Recherche Concertée to M.P. I.D.V and D.A.D. were supported by FNRS-Télévie grants and I.D.V by the Fondations Rose & Jean Hoguet and David & Alice Van Buuren, and O.D.H. was Aspirant of the Belgian Fonds National de la Recherche Scientifique.
Subjects: biology, Chemistry, [SDV]Life Sciences [q-bio], Lewis lung carcinoma, Cancer, tumor angiogenesis, Sciences bio-médicales et agricoles, CMKLR1, medicine.disease, Umbilical vein, Sciences humaines, ChemR23, Oncology, In vivo, biology.protein, Cancer research, medicine, Chemerin, Receptor, B16 melanoma, chemerin, Rarres2, Research Paper
Abstract: International audience; Chemerin, a multifunctional protein acting through the receptor ChemR23/CMKLR1, is downregulated in various human tumors and was shown to display antitumoral properties in mouse models of cancer. In the present study, we report that bioactive chemerin expression by tumor cells delays the growth of B16 melanoma and Lewis lung carcinoma in vivo. A similar delay is observed when chemerin is not expressed by tumor cells but by keratinocytes of the host mice. The protective effect of chemerin is mediated by CMKLR1 and appears unrelated to the recruitment of leukocyte populations. Rather, tumors grown in the presence of chemerin display a much smaller number of blood vessels, hypoxic regions early in their development, and larger necrotic areas. These observations likely explain the slower growth of the tumors. The anti-angiogenic effects of chemerin were confirmed in a bead sprouting assay using human umbilical vein endothelial cells. These results suggest that CMKLR1 agonists might constitute therapeutic molecules inhibiting the neoangiogenesis process in solid tumors.
Published: 2021
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9. Dynamics of myogenic differentiation using a novel Myogenin knock-in reporter mouse

Author: Benavente-Diaz, Maria, Comai, Glenda, Di Girolamo, Daniela, Langa, Francina, Tajbakhsh, Shahragim, Cellules Souches et Développement / Stem Cells and Development, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), ED 515 - Complexité du vivant, Sorbonne Université (SU), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), We acknowledge funding support from the Institut Pasteur, Agence Nationale de la Recherche (Laboratoire d’Excellence Revive, Investissement d’Avenir, ANR-10-LABX-73), Association Française contre les Myopathies (Grant #20510), and the Centre National de la Recherche Scientifique. M.B.D. was supported by a grant from Laboratoire d’Excellence Revive and La Ligue Contre le Cancer., ANR-10-LABX-0073,REVIVE,Stem Cells in Regenerative Biology and Medicine(2010), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], Benavente-Diaz, Maria, Comai, Glenda, Di Girolamo, Daniela, Langa, Francina, and Tajbakhsh, Shahragim
Subjects: lcsh:Diseases of the musculoskeletal system, [SDV]Life Sciences [q-bio], Methodology, Skeletal muscle, Cell Differentiation, Muscle Development, musculoskeletal system, Knock-in mouse, Myoblasts, Mice, Intravital imaging, Animals, Myogenin, lcsh:RC925-935, Muscle, Skeletal, tissues, [SDV.BDD]Life Sciences [q-bio]/Development Biology, tdTOMATO
Abstract: Background Myogenin is a transcription factor that is expressed during terminal myoblast differentiation in embryonic development and adult muscle regeneration. Investigation of this cell state transition has been hampered by the lack of a sensitive reporter to dynamically track cells during differentiation. Results Here, we report a knock-in mouse line expressing the tdTOMATO fluorescent protein from the endogenous Myogenin locus. Expression of tdTOMATO in MyogntdTom mice recapitulated endogenous Myogenin expression during embryonic muscle formation and adult regeneration and enabled the isolation of the MYOGENIN+ cell population. We also show that tdTOMATO fluorescence allows tracking of differentiating myoblasts in vitro and by intravital imaging in vivo. Lastly, we monitored by live imaging the cell division dynamics of differentiating myoblasts in vitro and showed that a fraction of the MYOGENIN+ population can undergo one round of cell division, albeit at a much lower frequency than MYOGENIN− myoblasts. Conclusions We expect that this reporter mouse will be a valuable resource for researchers investigating skeletal muscle biology in developmental and adult contexts. Supplementary Information The online version contains supplementary material available at 10.1186/s13395-021-00260-x.
Published: 2021
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10. Dissection of the Fgf8 regulatory landscape by in vivo CRISPR-editing reveals extensive intra- and inter-enhancer redundancy

Author: Katja Langenfeld, Francina Langa, S. Bastide, François Spitz, Andreas Hörnblad, European Molecular Biology Laboratory [Hamburg] (EMBL), Umeå University, Génomique et Epigénomique du Développement des Vertébrés - Genomics and Epigenomics of Vertebrates Development, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Université - Institut de Formation Doctorale (IFD ), Sorbonne Université (SU), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), A.H. was supported by an EMBL Interdisciplinary Postdoctoral fellowship (EIPOD) under Marie Curie Actions COFUND from the E.U. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (SP 1331/3-1, to F.S) and by the Institut Pasteur., Génomique et Epigénomique du Développement des Vertébrés - Genomics and Epigenomics of Vertebrate Development, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris]
Subjects: Male, 0301 basic medicine, Apical ectodermal ridge, MESH: CRISPR-Cas Systems, Transcriptional regulatory elements, Gene regulatory network, General Physics and Astronomy, Mice, 0302 clinical medicine, Mesencephalon, CRISPR, MESH: Embryonic Development, Gene Regulatory Networks, MESH: Animals, [SDV.BDD]Life Sciences [q-bio]/Development Biology, MESH: Gene Regulatory Networks, Regulation of gene expression, Multidisciplinary, Gene Expression Regulation, Developmental, MESH: Rhombencephalon, Enhancer Elements, Genetic, embryonic structures, MESH: Genetic Engineering, Female, Utvecklingsbiologi, animal structures, Lineage (genetic), Fibroblast Growth Factor 8, MESH: Mice, Transgenic, Science, Embryonic Development, Mice, Transgenic, Computational biology, Biology, Chromatin structure, Article, General Biochemistry, Genetics and Molecular Biology, Genome engineering, 03 medical and health sciences, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Ectoderm, Developmental biology, Animals, Enhancer, MESH: Mice, Cas9, MESH: Embryo, Mammalian, Extremities, MESH: Fibroblast Growth Factor 8, General Chemistry, MESH: Mesencephalon, Embryo, Mammalian, MESH: Male, Gene regulation, Rhombencephalon, 030104 developmental biology, Genetic engineering, Feasibility Studies, MESH: Enhancer Elements, Genetic, MESH: Extremities, CRISPR-Cas Systems, MESH: Feasibility Studies, 030217 neurology & neurosurgery, Developmental Biology
Abstract: Developmental genes are often regulated by multiple elements with overlapping activity. Yet, in most cases, the relative function of those elements and their contribution to endogenous gene expression remain poorly characterized. An example of this phenomenon is that distinct sets of enhancers have been proposed to direct Fgf8 in the limb apical ectodermal ridge and the midbrain-hindbrain boundary. Using in vivo CRISPR/Cas9 genome engineering, we functionally dissect this complex regulatory ensemble and demonstrate two distinct regulatory logics. In the apical ectodermal ridge, the control of Fgf8 expression appears distributed between different enhancers. In contrast, we find that in the midbrain-hindbrain boundary, one of the three active enhancers is essential while the other two are dispensable. We further dissect the essential midbrain-hindbrain boundary enhancer to reveal that it is also composed by a mixture of essential and dispensable modules. Cross-species transgenic analysis of this enhancer suggests that its composition may have changed in the vertebrate lineage., Developmental genes are often regulated by multiple elements, yet their relative contribution to gene expression remains poorly understood. Here the authors apply in vivo CRISPR/Cas9 genome engineering to find two distinct regulatory logics directing Fgf8 in the limb apical ectodermal ridge and the midbrain-hindbrain boundary.
Published: 2021
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11. Lysosomal Acid Lipase Drives Adipocyte Cholesterol Homeostasis and Modulates Lipid Storage in Obesity, Independent of Autophagy

Author: Emmanuel L. Gautier, Amélie Lacombe, Dominique Farabos, Christine Rouault, Camille Gamblin, Antonin Lamaziere, Laurent Yvan-Charvet, Francina Langa-Vives, Karine Clément, Isabelle Dugail, Nutrition et obésités: approches systémiques (UMR-S 1269) (Nutriomics), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Research Unit on Cardiovascular and Metabolic Diseases (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), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), Centre de Recherche Saint-Antoine (CRSA), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), CHU Saint-Antoine [AP-HP], 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), Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Research Unit on Cardiovascular and Metabolic Diseases [IHU ICAN], Institut Pasteur [Paris]-Université Paris Cité (UPCité), Gestionnaire, Hal Sorbonne Université, Nutrition et obésités: approches systémiques (nutriomics) (UMR-S 1269 INSERM - Sorbonne Université), 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], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Institut Pasteur [Paris], Centre de Recherche Saint-Antoine (CR Saint-Antoine), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Saint-Antoine [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), Université Nice Sophia Antipolis (... - 2019) (UNS), and COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: 0301 basic medicine, MESH: 3T3 Cells, Endocrinology, Diabetes and Metabolism, [SDV]Life Sciences [q-bio], Adipose tissue, Endoplasmic Reticulum, Mice, chemistry.chemical_compound, 0302 clinical medicine, MESH: Cholesterol, Adipocyte, Adipocytes, MESH: Obesity, MESH: Thermogenesis, MESH: Animals, MESH: Lipid Metabolism, MESH: Sterol Esterase, MESH: Middle Aged, Chemistry, Thermogenesis, 3T3 Cells, Middle Aged, [SDV] Life Sciences [q-bio], Cholesterol, medicine.anatomical_structure, Adult, MESH: Triglycerides, medicine.medical_specialty, Lipolysis, 030209 endocrinology & metabolism, 03 medical and health sciences, MESH: Endoplasmic Reticulum, Lysosome, Internal medicine, Autophagy, Internal Medicine, medicine, Animals, Humans, MESH: Autophagy, MESH: Lipolysis, Obesity, MESH: Mice, Triglycerides, MESH: Adipocytes, MESH: Humans, MESH: Adult, Sterol Esterase, Lipid Metabolism, Sterol regulatory element-binding protein, 030104 developmental biology, Endocrinology, Homeostasis
Abstract: International audience; Besides cytoplasmic lipase-dependent adipocyte fat mobilization, the metabolic role of lysosomal acid lipase (LAL), highly expressed in adipocytes, is unclear. We show that the isolated adipocyte fraction, but not the total undigested adipose tissue (ATs), from obese patients has decreased LAL expression compared with that from nonobese people. Lentiviral-mediated LAL knockdown in the 3T3L1 mouse cell line to mimic the obese adipocytes condition did not affect lysosome density or autophagic flux, but it did increase triglyceride storage and disrupt endoplasmic reticulum cholesterol, as indicated by activated SREBP. Conversely, mice with adipose-specific LAL overexpression (Adpn-rtTA x TetO-hLAL) gained less weight and body fat than did control mice fed a high-fat diet, resulting in ameliorated glucose tolerance. Blood cholesterol level in the former was lower than that of control mice, although triglyceridemia in the two groups of mice was similar. The adipose-specific LAL-overexpressing mouse phenotype depends on the housing temperature and develops only under mild hypothermic stress (e.g., room temperature) but not at thermoneutrality (30°C), demonstrating the prominent contribution of brown AT (BAT) thermogenesis. LAL overexpression increased levels of BAT free cholesterol, decreased SREBP targets, and induced the expression of genes involved in initial steps of mitochondrial steroidogenesis, suggesting conversion of lysosome-derived cholesterol to pregnenolone. In conclusion, our study demonstrates that adipose LAL drives tissue-cholesterol homeostasis and affects BAT metabolism, suggesting beneficial LAL activation in anti-obesity approaches aimed at reactivating thermogenic energy expenditure.
Published: 2020
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12. Corrigendum: Expression of Bioactive Chemerin by Keratinocytes Inhibits Late Stages of Tumor Development in a Chemical Model of Skin Carcinogenesis

Author: Ingrid Dubois-Vedrenne, Olivier De Henau, Virginie Robert, Francina Langa, Joaquim Javary, Diana Al Delbany, Olivier Vosters, Edgar Angelats-Canals, Maxime Vernimmen, Souphalone Luangsay, Valérie Wittamer, Marc Parmentier, Institut de Recherche Interdisciplinaire en Biologie Humaine et moléculaire = Insitute of Interdisciplinary Research [Bruxelles, Belgique] (IRIBHM), Faculté de Médecine [Bruxelles] (ULB), Université libre de Bruxelles (ULB)-Université libre de Bruxelles (ULB), Walloon Excellence in Life sciences and BIOtechnology [Liège] (WELBIO), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], Ogeda [Gosselies, Belgique], This work was supported by the Belgian programme on Interuniversity Poles of Attraction initiated by the Belgian State, Prime Minister's Office, Science Policy Programming, the Fonds National de la Recherche Scientifique and Welbio (grant CR-2017C-03), Télévie, the Fondation Belge contre le Cancer and the Actions de Recherche Concertée to MP. ID-V and DA were supported by FNRS-Télévie grants and ID-V by the Fondations Rose & Jean Hoguet and David & Alice Van Buuren, OD was Aspirant of the Belgian Fonds National de la Recherche Scientifique., and Institut Pasteur [Paris] (IP)
Subjects: squamous cell carcinoma, 0301 basic medicine, Cancer Research, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, CMKLR1, medicine.disease_cause, lcsh:RC254-282, ChemR23, chemical carcinogenesis, 03 medical and health sciences, Basal (phylogenetics), 0302 clinical medicine, medicine, Chemerin, Receptor, Original Research, Melanoma, Correction, Cancer, Sciences bio-médicales et agricoles, lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens, medicine.disease, 3. Good health, 030104 developmental biology, Oncology, Tumor progression, 030220 oncology & carcinogenesis, Cancer research, biology.protein, Carcinogenesis, chemerin
Abstract: Chemerin is a multifunctional protein acting mainly through the G protein-coupled receptor ChemR23/CMKLR1/Chemerin1. Its expression is frequently downregulated in human tumors, including in melanoma and squamous cell carcinoma of the skin and anti-tumoral properties of chemerin were reported in mouse tumor graft models. In the present study, we report the development of spontaneous skin tumors in aged ChemR23-deficient mice. In order to test the potential therapeutic benefit of chemerin analogs, a transgenic model in which bioactive chemerin is over-expressed by basal keratinocytes was generated. These animals are characterized by increased levels of chemerin immunoreactivity and bioactivity in the skin and the circulation. In a chemical carcinogenesis model, papillomas developed later, were less numerous, and their progression to carcinomas was delayed. Temporal control of chemerin expression by doxycycline allowed to attribute its effects to late stages of carcinogenesis. The protective effects of chemerin were partly abrogated by ChemR23 invalidation. These results demonstrate that chemerin is able to delay very significantly tumor progression in a model that recapitulates closely the evolution of solid cancer types in human and suggest that the chemerin-ChemR23 system might constitute an interesting target for therapeutic intervention in the cancer field., info:eu-repo/semantics/published
Published: 2020
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13. Mitochondrial AIF Loss Causes Metabolic Reprogramming, Caspase-Independent Cell Death Blockade, Embryonic Lethality, and Perinatal Hydrocephalus

Author: Francina Langa-Vives, Christophe Lemaire, Delphine Garnier, Mariana Tannoury, Mathieu Baritaud, Raquel Moreno-Loshuertos, Martine Cohen-Salmon, Kevin Garbin, Laure Delavallée, Fabrice Chrétien, Lauriane Cabon, Patricio Fernández-Silva, Navrita Mathiah, Marie-Noëlle Brunelle-Navas, Laura Vela, Leticia K. Lerner, Santos A. Susin, Isabelle Migeotte, Alexandre Prola, Aurélien Mazeraud, 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), Institut de Recherche Interdisciplinaire en Biologie Humaine et moléculaire = Insitute of Interdisciplinary Research [Bruxelles, Belgique] (IRIBHM), Faculté de Médecine [Bruxelles] (ULB), Université libre de Bruxelles (ULB)-Université libre de Bruxelles (ULB), Neuropathologie expérimentale - Experimental neuropathology, Institut Pasteur [Paris]-Université Paris Descartes - Paris 5 (UPD5), Centre Hospitalier Sainte Anne [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Signalisation et physiopathologie cardiovasculaire (CARPAT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay, Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), University of Zaragoza - Universidad de Zaragoza [Zaragoza], Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], Centre interdisciplinaire de recherche en biologie (CIRB), Labex MemoLife, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), This work was supported by Fondation ARC (PJA20151203407 and PJA20171206551), Fondation pour la Recherche Médicale, French National Research Agency (ANR-09-BLAN-0247), and the SIRIC-CURAMUS (INCA-DGOS-Inserm_12560). L.C. received PhD fellowship support from ENS-Cachan, Société Francophone du Diabète, and Fondation ARC. N.M. received a fellowship from the FRS/FRIA. I.M. is an FNRS research associate and an investigator of WELBIO., ANR-09-BLAN-0247,AIFCICD(2009), Institut Pasteur [Paris] (IP)-Université Paris Descartes - Paris 5 (UPD5), Institut Pasteur [Paris] (IP), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), LANGA VIVES, FRANCINA, Blanc - - AIFCICD2009 - ANR-09-BLAN-0247 - Blanc - VALID, and École Pratique des Hautes Études (EPHE)
Subjects: 0301 basic medicine, Male, Cellular differentiation, Apoptosis, Mitochondrion, Oxidative Phosphorylation, Mice, AIF, 0302 clinical medicine, Caspase-independent cell death, 2. Zero hunger, Membrane Potential, Mitochondrial, Hydrocephaly, Apoptosis Inducing Factor, Sciences bio-médicales et agricoles, OXPHOS, Cell biology, Mitochondria, Caspases, Knockout mouse, Models, Animal, Female, biological phenomena, cell phenomena, and immunity, Genetic Engineering, Glycolysis, Hydrocephalus, Senescence, Programmed cell death, lcsh:Internal medicine, Cell Respiration, 030209 endocrinology & metabolism, Mice, Inbred Strains, Oxidative phosphorylation, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Article, 03 medical and health sciences, Animals, cardiovascular diseases, lcsh:RC31-1245, Molecular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Cell Biology, Fibroblasts, Mice, Inbred C57BL, 030104 developmental biology, Metabolism, Anaerobic glycolysis, Reactive Oxygen Species
Abstract: Objectives Apoptosis-Inducing Factor (AIF) is a protein involved in mitochondrial electron transport chain assembly/stability and programmed cell death. The relevant role of this protein is underlined because mutations altering mitochondrial AIF properties result in acute pediatric mitochondriopathies and tumor metastasis. By generating an original AIF-deficient mouse strain, this study attempted to analyze, in a single paradigm, the cellular and developmental metabolic consequences of AIF loss and the subsequent oxidative phosphorylation (OXPHOS) dysfunction. Methods We developed a novel AIF-deficient mouse strain and assessed, using molecular and cell biology approaches, the cellular, embryonic, and adult mice phenotypic alterations. Additionally, we conducted ex vivo assays with primary and immortalized AIF knockout mouse embryonic fibroblasts (MEFs) to establish the cell death characteristics and the metabolic adaptive responses provoked by the mitochondrial electron transport chain (ETC) breakdown. Results AIF deficiency destabilized mitochondrial ETC and provoked supercomplex disorganization, mitochondrial transmembrane potential loss, and high generation of mitochondrial reactive oxygen species (ROS). AIF-/Y MEFs counterbalanced these OXPHOS alterations by mitochondrial network reorganization and a metabolic reprogramming toward anaerobic glycolysis illustrated by the AMPK phosphorylation at Thr172, the overexpression of the glucose transporter GLUT-4, the subsequent enhancement of glucose uptake, and the anaerobic lactate generation. A late phenotype was characterized by the activation of P53/P21-mediated senescence. Notably, approximately 2% of AIF-/Y MEFs diminished both mitochondrial mass and ROS levels and spontaneously proliferated. These cycling AIF-/Y MEFs were resistant to caspase-independent cell death inducers. The AIF-deficient mouse strain was embryonic lethal between E11.5 and E13.5 with energy loss, proliferation arrest, and increased apoptotic levels. Contrary to AIF-/Y MEFs, the AIF KO embryos were unable to reprogram their metabolism toward anaerobic glycolysis. Heterozygous AIF+/- females displayed progressive bone marrow, thymus, and spleen cellular loss. In addition, approximately 10% of AIF+/- females developed perinatal hydrocephaly characterized by brain development impairment, meningeal fibrosis, and medullar hemorrhages; those mice died 5 weeks after birth. AIF+/- with hydrocephaly exhibited loss of ciliated epithelium in the ependymal layer. This phenotype was triggered by the ROS excess. Accordingly, it was possible to diminish the occurrence of hydrocephalus AIF+/- females by supplying dams and newborns with an antioxidant in drinking water. Conclusions In a single knockout model and at 3 different levels (cell, embryo, and adult mice) we demonstrated that by controlling the mitochondrial OXPHOS/metabolism, AIF is a key factor regulating cell differentiation and fate. Additionally, by providing new insights into the pathological consequences of mitochondrial OXPHOS dysfunction, our new findings pave the way for novel pharmacological strategies., Highlights • Mitochondrial AIF loss triggers OXPHOS/metabolism impairment. • AIF-/Y MEFs reprogram their metabolism by enabling an AMPK/GLUT-4 glycolytic pathway. • OXPHOS dysfunction associated with AIF loss generates a P53/P21 senescent phenotype. • The lack of metabolic plasticity provokes embryonic lethality in the AIF KO animals. • AIF+/- females develop immune cell exhaustion and perinatal hydrocephaly.
Published: 2020
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14. 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
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15. A distinct cardiopharyngeal mesoderm genetic hierarchy establishes antero-posterior patterning of esophagus striated muscle

Author: Francesca Pala, Shahragim Tajbakhsh, Francina Langa, Glenda Comai, Gabrielle Kardon, Sebastian Mella, Sylvain Paisant, Eglantine Heude, Swetha Gopalakrishnan, Mirialys Gallardo, Cellules Souches et Développement / Stem Cells and Development, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Département Adaptations du vivant (AVIV), Muséum national d'Histoire naturelle (MNHN), Laboratory of Clinical Immunology and Microbiology [Bethesda, MA, USA] (LCIM), National Institute of Allergy and Infectious Diseases [Bethesda] (NIAID-NIH), National Institutes of Health [Bethesda] (NIH)-National Institutes of Health [Bethesda] (NIH), Department of Human Genetics [Salt Lake City, UT, USA], University of Utah, Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], HiLIFE - Neuroscience Center (NC), Helsinki Institute of Life Science (HiLIFE), University of Helsinki-University of Helsinki, We acknowledge funding support from the Institut Pasteur, Association Franc¸ aise contre le Myopathies, Agence Nationale de la Recherche (Laboratoire d’Excellence Revive, Investissement d’Avenir, ANR-10-LABX-73)., ANR-10-LABX-0073,REVIVE,Stem Cells in Regenerative Biology and Medicine(2010), Heude, Eglantine, Laboratoires d'excellence - Stem Cells in Regenerative Biology and Medicine - - REVIVE2010 - ANR-10-LABX-0073 - LABX - VALID, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Physiologie moléculaire et adaptation (PhyMA), Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), Helsingin yliopisto = Helsingfors universitet = University of Helsinki-Helsingin yliopisto = Helsingfors universitet = University of Helsinki, Centre of Excellence in Stem Cell Metabolism, and Institute of Biotechnology
Subjects: Mouse, Tbx1-Isl1-Met/HGF, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Mesoderm, Mice, 0302 clinical medicine, SKELETAL MYOGENESIS, Biology (General), [SDV.BDD]Life Sciences [q-bio]/Development Biology, 0303 health sciences, Hepatocyte Growth Factor, Myogenesis, General Neuroscience, C-MET RECEPTOR, 1184 Genetics, developmental biology, physiology, Gene Expression Regulation, Developmental, General Medicine, Proto-Oncogene Proteins c-met, Cell biology, DIFFERENTIATION, medicine.anatomical_structure, Medicine, MOUSE ESOPHAGUS, Signal Transduction, Research Article, EXPRESSION, TBX1, GROWTH-FACTOR, QH301-705.5, PHARYNGEAL ARCH, Science, LIM-Homeodomain Proteins, Biology, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Esophagus, REVEALS, [SDV.BDD] Life Sciences [q-bio]/Development Biology, myogenic migration, medicine, Animals, HEAD, Body Patterning, 030304 developmental biology, Progenitor, Evolutionary Biology, General Immunology and Microbiology, antero-posterior patterning, cardiopharyngeal mesoderm, Skeletal muscle, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Muscle, Striated, PROGENITOR CELLS, esophagus striated muscles, ISL1, T-Box Domain Proteins, 030217 neurology & neurosurgery, Pharyngeal arch, Transcription Factors, Developmental Biology
Abstract: International audience; In most vertebrates, the upper digestive tract is composed of muscularized jaws linked to the esophagus that permits food ingestion and swallowing. Masticatory and esophagus striated muscles (ESM) share a common cardiopharyngeal mesoderm (CPM) origin, however ESM are unusual among striated muscles as they are established in the absence of a primary skeletal muscle scaffold. Using mouse chimeras, we show that the transcription factors Tbx1 and Isl1 are required cell-autonomously for myogenic specification of ESM progenitors. Further, genetic loss-of-function and pharmacological studies point to MET/HGF signaling for antero-posterior migration of esophagus muscle progenitors, where Hgf ligand is expressed in adjacent smooth muscle cells. These observations highlight the functional relevance of a smooth and striated muscle progenitor dialogue for ESM patterning. Our findings establish a Tbx1-Isl1-Met genetic hierarchy that uniquely regulates esophagus myogenesis and identify distinct genetic signatures that can be used as framework to interpret pathologies arising within CPM derivatives.
Published: 2019
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16. 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
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17. A Human Polymorphism in CHRNA5 Is Linked to Relapse to Nicotine Seeking in Transgenic Rats

Author: Francina Langa, Uwe Maskos, Morgane Besson, Alexandre Mourot, Carole Morel, Sarah Mondoloni, Romain Durand-de Cuttoli, Bertrand Lambolez, Petra Scholze, Stéphanie Pons, Ludovic Tricoire, Audrey Hay, Benoit Forget, Philippe Faure, Neurobiologie intégrative des Systèmes cholinergiques / Integrative Neurobiology of Cholinergic Systems (NISC), Institut Pasteur [Paris] (IP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Center for Brain Research [Vienna, Austria], Medizinische Universität Wien = Medical University of Vienna, Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), Neurobiologie des processus adaptatifs (NPA), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Neurophysiologie et comportements = Neurophysiology and Behavior (NPS-06), Neuroscience Paris Seine (NPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Biologie Paris Seine (IBPS), 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)-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)-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)-Institut de Biologie Paris Seine (IBPS), 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)-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)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Réseau cortical et couplage neurovasculaire = Cortical Network and Neurovascular (NPS-03), Neurophysiologie et comportements = Neurophysiology and Behavior (NPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), 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 (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), 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 (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), This work was supported by the Institut Pasteur, Centre National de la Recherche Scientifique UMR 3571 and 8246, the programs PasteurInnov2012 and PasteurInnov 2013, the Fondation de la Recherche Médicale (FRM) (grants SPF20140129365 and DPA20140629803 to U.M.), the Agence Nationale de la Recherche (ANR), ANR Neuroscience and ANR BLANC,Laboratoire d’Excellence LABEX BIO-PSY by the FP7 ERANET program NICO-GENE grant agreement convention ANR no. 2010-NEUR-004-01, the European Commission FP7 RTD Project HEALTH-2009-Neurocyp.08-202088 grant 242167, and French National Cancer Institute grant CANCEROPOLE IDF 2016-1-TABAC-01-IP-1 MASKOS (all to U.M.). This work was also supported by the ANR-JC JC Nicopto, a NARSAD Young Investigator Grant from the Brain and Behavior Research Foundation, and the Medisite Foundation to A.M. R.D.C. was supported by a Ph.D. fellowship from the DIM Cerveau and Pensee program of the Region Ile-de-France. The laboratories of U.M. and P.F. are part of the Ecole des Neurosciences de Paris Ile-de-France RTRA network. U.M. and P.F. are members of the LABEX BIO-PSY. As such, this work was supported by French state funds managed by the ANR within the Investissements d’Avenir programme under reference ANR-11-IDEX-0004-02., ANR-11-IDEX-0004,SUPER,Sorbonne Universités à Paris pour l'Enseignement et la Recherche(2011), ANR-06-NEUR-0043,Presynaptic-CB1,Récepteurs présynaptique cannabinoïdes de type 1 (CB1): mécanismes et fonctions dans le cortex préfrontal.(2006), European Project: 202088,EC:FP7:HEALTH,FP7-HEALTH-2007-A,NEUROCYPRES(2008), European Project: 242167,EC:FP7:HEALTH,FP7-HEALTH-2009-two-stage,SYNSYS(2010), European Project: 680966,ERA-NET NEURON,NICO-GENE(2015), Sorbonne Université (SU)-Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Center for Brain Research [Vienne], Institut Pasteur [Paris], Neurosciences Paris Seine (NPS), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Biologie Paris Seine (IBPS), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-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)-Institut de Biologie Paris Seine (IBPS), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-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), Neurobiologie intégrative des Systèmes cholinergiques - Integrative Neurobiology of Cholinergic Systems, Institut Pasteur [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC), Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), ANR-11-IDEX-0004-02/11-LABX-0035,BIOPSY,Laboratoire de Psychiatrie Biologique(2011), and ANR-06-NEURO-0043,PRESYNAPTIC-CB1,Récepteurs présynaptique cannabinoïdes de type 1 (CB1): mécanismes et fonctions dans le cortex préfrontal.(2006)
Subjects: 0301 basic medicine, Male, medicine.medical_specialty, Interpeduncular nucleus, Nicotine, medicine.medical_treatment, media_common.quotation_subject, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Self Administration, Biology, Receptors, Nicotinic, Polymorphism, Single Nucleotide, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, 0302 clinical medicine, Recurrence, Internal medicine, medicine, Premovement neuronal activity, Animals, Humans, Rats, Long-Evans, ComputingMilieux_MISCELLANEOUS, media_common, Acetylcholine receptor, Base Sequence, CHRNA5, Addiction, Tobacco Use Disorder, 3. Good health, Rats, 030104 developmental biology, Endocrinology, Nicotinic agonist, biology.protein, Smoking cessation, Female, Rats, Transgenic, General Agricultural and Biological Sciences, Sequence Alignment, 030217 neurology & neurosurgery, medicine.drug
Abstract: Summary Tobacco addiction is a chronic and relapsing disorder with an important genetic component that represents a major public health issue. Meta-analysis of large-scale human genome-wide association studies (GWASs) identified a frequent non-synonymous SNP in the gene coding for the α5 subunit of nicotinic acetylcholine receptors (α5SNP), which significantly increases the risk for tobacco dependence and delays smoking cessation. To dissect the neuronal mechanisms underlying the vulnerability to nicotine addiction in carriers of the α5SNP, we created rats expressing this polymorphism using zinc finger nuclease technology and evaluated their behavior under the intravenous nicotine-self-administration paradigm. The electrophysiological responses of their neurons to nicotine were also evaluated. α5SNP rats self-administered more nicotine at high doses and exhibited higher nicotine-induced reinstatement of nicotine seeking than wild-type rats. Higher reinstatement was associated with altered neuronal activity in several discrete areas that are interconnected, including in the interpeduncular nucleus (IPN), a GABAergic structure that strongly expresses α5-containing nicotinic receptors. The altered reactivity of IPN neurons of α5SNP rats to nicotine was confirmed electrophysiologically. In conclusion, the α5SNP polymorphism is a major risk factor for nicotine intake at high doses and for relapse to nicotine seeking in rats, a dual effect that reflects the human condition. Our results also suggest an important role for the IPN in the higher relapse to nicotine seeking observed in α5SNP rats.
Published: 2018
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18. Reduced recruitment of 53BP1 during interstrand crosslink repair is associated with genetically inherited attenuation of mitomycin C sensitivity in a family with Fanconi anemia

Author: Francina Langa Vives, Jean Soulier, Armand Biver, Jean-Pierre de Villartay, Patrick Revy, Nada Jabado, Benoit Roch, Emilie Lesport, Nadia Vasquez, Alina Ferster, LANGA VIVES, FRANCINA, Programme de Recherche Translationnelle en Santé - Conséquences des défauts de réparation de l'ADN et du maintien des télomères sur le fonctionnement du Système Immunitaire: application aux DICV et aux déficits immunitaires avec syndromes dysmorphiques - - Immune-Rep2013 - ANR-13-PRTS-0004 - PRTS - VALID, Primary immunodeficiency diseases as models for the study of the immune system - PIDIMMUN - - EC:FP7:ERC2010-07-01 - 2015-06-30 - 249816 - VALID, 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 Paris Cité (UPCité)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Hôpital Universitaire des Enfants Reine Fabiola [Bruxelles, Belgique] (HUDERF), Centre Hospitalier de Luxembourg [Luxembourg] (CHL), Génomes, biologie cellulaire et thérapeutiques (GenCellDi (U944 / UMR7212)), Collège de France (CdF (institution))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), McGill University = Université McGill [Montréal, Canada], Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), This work was supported by institutional grants from INSERM, the Institut National du Cancer (PLBIO11-151) and Agence Nationale de la Recherche (ANR-13-PRTS-0004), by grants from La Ligue Nationale contre le Cancer (Equipe Labellisée LA LIGUE), and ERC PID-IMMUN (no. 249816). EL was supported by a grant from ARC Fondation., ANR-13-PRTS-0004,Immune-Rep,Conséquences des défauts de réparation de l'ADN et du maintien des télomères sur le fonctionnement du Système Immunitaire: application aux DICV et aux déficits immunitaires avec syndromes dysmorphiques(2013), European Project: 249816,EC:FP7:ERC,ERC-2009-AdG,PIDIMMUN(2010), 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), Génomes, biologie cellulaire et thérapeutiques (GenCellDi (UMR_S_944)), Collège de France (CdF (institution))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), and Institut Pasteur [Paris]
Subjects: 0301 basic medicine, [SDV]Life Sciences [q-bio], Interstrand crosslink, Biology, [SDV.GEN.GH] Life Sciences [q-bio]/Genetics/Human genetics, 03 medical and health sciences, chemistry.chemical_compound, FANCG, Fanconi anemia, medicine, Mitomycin C, DNA interstrand crosslinks, medicine.disease, Phenotype, 53BP1, Double Strand Break Repair, [SDV] Life Sciences [q-bio], 030104 developmental biology, Oncology, chemistry, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, DNA double strand break repair, ATM, Cancer research, Homologous recombination, DNA, Research Paper, Psychiatrie
Abstract: The Fanconi anemia (FA) pathway is implicated in the repair of DNA interstrand crosslinks (ICL). In this process, it has been shown that FA factors regulate the choice for DNA double strand break repair towards homologous recombination (HR). As this mechanism is impaired in FA deficient cells exposed to crosslinking agents, an inappropriate usage of non-homologous end joining (NHEJ) leads to the accumulation of toxic chromosomal abnormalities. We studied a family with two FANCG patients and found a genetically inherited attenuation of mitomycin C sensitivity resulting in-vitro in an attenuated phenotype for one patient or in increased resistance for two healthy relatives. A heterozygous mutation in ATM was identified in these 3 subjects but was not directly linked to the observed phenotype. However, the attenuation of ICL sensitivity was associated with a reduced recruitment of 53BP1 during the course of ICL repair, and increased HR levels. These results further demonstrate the importance of favoring HR over NHEJ for the survival of cells challenged with ICLs., SCOPUS: ar.j, info:eu-repo/semantics/published
Published: 2018
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19. Penicillin Resistance Compromises Nod1-Dependent Proinflammatory Activity and Virulence Fitness of Neisseria meningitidis

Author: Francina Langa Vives, Aude Antignac, Muhamed-Kheir Taha, Ala-Eddine Deghmane, Anna Skoczynska, Faridabano Nato, Philippe J. Sansonetti, Stephen E. Girardin, Ivo G. Boneca, Lucie Peduto, Françoise Thouron, Martine Fanton d'Andon, Thierry Pedron, Meriem El Ghachi, Marek Szatanik, Dana J. Philpott, Catherine Werts, Lionel LeBourhis, Maria Leticia Zarantonelli, Céline Mulet, Gérard Eberl, Neisseria, Institut Pasteur [Paris], National Reference Centre for Bacterial Meningitis [Warsaw, Poland] (NRCBM), National Medicines Institute - Narodowy Instytut Leków [Warsaw] (NIL), Infections Bactériennes Invasives, Biologie et Génétique de la Paroi bactérienne - Biology and Genetics of Bacterial Cell Wall (BGPB), Groupe Avenir, Institut National de la Santé et de la Recherche Médicale (INSERM), Pathogénie Microbienne Moléculaire, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Développement des Tissus Lymphoïdes, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Production de Protéines Recombinantes et d'Anticorps (Plate-Forme), Immunité Innée et Signalisation, Chaire Microbiologie et Maladies infectieuses, Collège de France (CdF (institution)), Our research is supported by Institut Pasteur grants (PTR153 and PTR187), an ACI 0321 (Action Concertée Incitative Intégrée Microbiology) Grant from the Ministère chargé de la Recherche (INSERM No. MIC 0321), and a European Research Council starting grant (202283-PGNfromSHAPEtoVIR). A.S. was supported by a Marie Curie Intra-European Fellowship (No. 23188) within the Sixth European Community Framework Programme and Marie Curie European Reintegration Grants (No. 234876) within the Seventh European Community Framework Programme. M.E.G. was supported by an Institut Pasteur Roux fellowship., European Project: 234876,EC:FP7:PEOPLE,FP7-PEOPLE-ERG-2008,NMEN-PBP(2009), Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Collège de France - Chaire Microbiologie et Maladies infectieuses
Subjects: Cancer Research, Penicillin Resistance, Virulence, MESH: Nod1 Signaling Adaptor Protein, Biology, Neisseria meningitidis, medicine.disease_cause, Microbiology, MESH: Neisseria meningitidis, Proinflammatory cytokine, Sepsis, 03 medical and health sciences, chemistry.chemical_compound, Mice, MESH: Mutant Proteins, Antibiotic resistance, MESH: Cell Wall, Cell Wall, Virology, Nod1 Signaling Adaptor Protein, Immunology and Microbiology(all), medicine, Animals, Humans, Penicillin-Binding Proteins, MESH: Animals, Pathogen, MESH: Mice, Molecular Biology, 030304 developmental biology, 0303 health sciences, MESH: Penicillin-Binding Proteins, MESH: Humans, 030306 microbiology, medicine.disease, MESH: Penicillin Resistance, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, 3. Good health, Penicillin, chemistry, Parasitology, Mutant Proteins, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, Peptidoglycan, medicine.drug
Abstract: International audience; Neisseria meningitidis is a life-threatening human bacterial pathogen responsible for pneumonia, sepsis, and meningitis. Meningococcal strains with reduced susceptibility to penicillin G (Pen(I)) carry a mutated penicillin-binding protein (PBP2) resulting in a modified peptidoglycan structure. Despite their antibiotic resistance, Pen(I) strains have failed to expand clonally. We analyzed the biological consequences of PBP2 alteration among clinical meningococcal strains and found that peptidoglycan modifications of the Pen(I) strain resulted in diminished in vitro Nod1-dependent proinflammatory activity. In an influenza virus-meningococcal sequential mouse model mimicking human disease, wild-type meningococci induced a Nod1-dependent inflammatory response, colonizing the lungs and surviving in the blood. In contrast, isogenic Pen(I) strains were attenuated for such response and were out-competed by meningococci sensitive to penicillin G. Our results suggest that antibiotic resistance imposes a cost to the success of the pathogen and may potentially explain the lack of clonal expansion of Pen(I) strains.
Published: 2013
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20. MITF – A controls branching morphogenesis and nephron endowment

Author: Phelep, Aurélie, Laouari, Denise, Bharti, Kapil, Burtin, Martine, Tammaccaro, Salvina, Garbay, Serge, Nguyen, Clement, Vasseur, Florence, Blanc, Thomas, Berissi, Sophie, Langa-Vives, Francina, Fischer, Evelyne, Druilhe, Anne, Arnheiter, Heinz, Friedlander, Gerard, Pontoglio, Marco, Terzi, Fabiola, Bodescot, Myriam, BLANC - Signatures moléculaires de la progression des maladies rénales chroniques - - SIGNA-KID2011 - ANR-11-BSV1-0029 - BLANC - VALID, Le rôle divergent d'EGF et TGFa dans les maladies rénales chroniques : vers des nouvelles cibles thérapeutiques - - GiF-KID2016 - ANR-16-CE14-0023 - AAPG2016 - VALID, Université Sorbonne Paris Cité - - USPC2011 - ANR-11-IDEX-0005 - IDEX - VALID, Département Croissance et Signalisation [Paris], Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), 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), Ocular and Stem Cell Translational Research Unit [Bethesda, MD, États-Unis], National Eye Institute [Bethesda, MD, États-Unis] (NEI), National Institutes of Health [Bethesda] (NIH)-National Institutes of Health [Bethesda] (NIH), 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), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), National Institute of Neurological Disorders and Stroke [Bethesda] (NINDS), National Institutes of Health [Bethesda] (NIH), This work was supported by INSERM, Université Paris Descartes - Sorbonne Paris Cité, Université Paris Pierre et Marie Curie, Agence Nationale de la Recherche (ANR-11-BSV1-0029, ANR-16-CE14-0023-01), Fondation de la Recherche Médicale (DEQ20061107968), WHOAMI Labex ANR (ANR-11-LABX-0071, Idex ANR-11-IDEX-0005-02)., ANR-11-BSV1-0029,SIGNA-KID,Signatures moléculaires de la progression des maladies rénales chroniques(2011), ANR-16-CE14-0023,GiF-KID,Le rôle divergent d'EGF et TGFa dans les maladies rénales chroniques : vers des nouvelles cibles thérapeutiques(2016), ANR-11-IDEX-0005,USPC,Université Sorbonne Paris Cité(2011), Institut Pasteur [Paris], ANR-16-CE14-0023,GiF-KID,Le rôle divergent d’EGF et TGFa dans les maladies rénales chroniques : vers des nouvelles cibles thérapeutiques(2016), and ANR-11-IDEX-0005-02/11-LABX-0071,WHO AM I,Determinants de l’Identité : de la molécule à l’individu(2011)
Subjects: Male, Embryology, lcsh:QH426-470, Organogenesis, Gene Expression, Mouse Models, Mice, Transgenic, Research and Analysis Methods, Kidney, urologic and male genital diseases, Mice, Model Organisms, Branching Morphogenesis, [SDV.BDD] Life Sciences [q-bio]/Development Biology, Medicine and Health Sciences, Genetics, Morphogenesis, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Animals, Humans, Protein Isoforms, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Microphthalmia-Associated Transcription Factor, Genetically Modified Animals, integumentary system, urogenital system, Genetically Modified Organisms, Embryos, Proto-Oncogene Proteins c-ret, Biology and Life Sciences, Kidneys, Renal System, Nephrons, Animal Models, lcsh:Genetics, Experimental Organism Systems, Female, Anatomy, Ureter, Genetic Engineering, Glomeruli, Research Article, Developmental Biology, Biotechnology
Abstract: Congenital nephron number varies widely in the human population and individuals with low nephron number are at risk of developing hypertension and chronic kidney disease. The development of the kidney occurs via an orchestrated morphogenetic process where metanephric mesenchyme and ureteric bud reciprocally interact to induce nephron formation. The genetic networks that modulate the extent of this process and set the final nephron number are mostly unknown. Here, we identified a specific isoform of MITF (MITF-A), a bHLH-Zip transcription factor, as a novel regulator of the final nephron number. We showed that overexpression of MITF-A leads to a substantial increase of nephron number and bigger kidneys, whereas Mitfa deficiency results in reduced nephron number. Furthermore, we demonstrated that MITF-A triggers ureteric bud branching, a phenotype that is associated with increased ureteric bud cell proliferation. Molecular studies associated with an in silico analyses revealed that amongst the putative MITF-A targets, Ret was significantly modulated by MITF-A. Consistent with the key role of this network in kidney morphogenesis, Ret heterozygosis prevented the increase of nephron number in mice overexpressing MITF-A. Collectively, these results uncover a novel transcriptional network that controls branching morphogenesis during kidney development and identifies one of the first modifier genes of nephron endowment., Author summary The number of nephrons, the functional unit of kidney, varies widely among humans. Indeed, it has been shown that kidneys may contain from 0.3 to more than 2 million of nephrons. Nephrons are formed during development via a coordinated morphogenetic program in which the metanephric mesenchyme reciprocally and recursively interacts with the ureteric bud. The fine-tuning of this cross-talk determines the final number of nephrons. Strong evidence indicates that suboptimal nephron endowment is associated with an increased risk of hypertension and chronic kidney disease, a major healthcare burden. Indeed, chronic kidney disease is characterized by the progressive decline of renal function towards end stage renal disease, which occurs once a critical number of nephrons has been lost. Elucidating the molecular mechanisms that control nephron endowment is, therefore, a critical issue for public health. However, little is known about the factors that determine the final number of nephrons in the healthy population. Our data showed that nephron endowment is genetically predetermined and identified Mitfa, a bHLH transcription factor, as one of the first modifiers of nephron formation during kidney development. By generating an allelic series of transgenic mice expressing different levels of MITF-A, we discovered that MITF-A promotes final nephron endowment. In addition, we elucidated the molecular mechanisms by which MITF-A promotes nephron formation and identified RET as one of the critical effectors.
Published: 2017
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21. Generation of a Double KO Mouse by Simultaneous Targeting of the Neighboring Genes Tmem176a and Tmem176b Using CRISPR/Cas9: Key Steps from Design to Genotyping

Author: Gaëlle Chauveau-Le Friec, Francina Langa, Aurélie Lemoine, Cédric Louvet, Centre de Recherche en Transplantation et Immunologie (U1064 Inserm - CRTI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), Institut de transplantation urologie-néphrologie (ITUN), Université de Nantes (UN)-Centre hospitalier universitaire de Nantes (CHU Nantes), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], Nantes Métropole, Région Pays de la Loire and Fondation Progreffe., ANR-11-LABX-0016,IGO,Immunothérapies Grand Ouest(2011), ANR-10-IBHU-0005,CESTI (TSI-IHU),Centre Européen des Sciences de la Transplantation et de l'Immunothérapie (TSI-IHU)(2010), Le Bihan, Sylvie, Laboratoires d'excellence - Immunothérapies Grand Ouest - - IGO2011 - ANR-11-LABX-0016 - LABX - VALID, Instituts Hospitalo-Universitaires B - Centre Européen des Sciences de la Transplantation et de l'Immunothérapie (TSI-IHU) - - CESTI (TSI-IHU)2010 - ANR-10-IBHU-0005 - IBHU - VALID, and Institut Pasteur [Paris] (IP)
Subjects: 0301 basic medicine, Genotyping, DNA End-Joining Repair, Genotyping Techniques, C57BL/6, Locus (genetics), Biology, Deletion, Gene Knockout Techniques, Mice, 03 medical and health sciences, Genome editing, Genetics, Animals, CRISPR, Molecular Biology, Gene, CRISPR/Cas9, Double KO, NHEJ, Mice, Knockout, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, Base Sequence, Cas9, Mosaicism, Inversion, Membrane Proteins, Founder Effect, Off-targets, 030104 developmental biology, CRISPR-Cas Systems, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: International audience; The CRISPR/Cas9 system has been tailored to a revolutionary genetic tool because of its remarkable simplicity and efficacy. While complex genome editing in the mouse since the 1990s has been dominated by the use of embryonic stem (ES) cells, CRISPR/Cas9 now offers a versatile and fast approach to precisely modify virtually any DNA regions directly in mouse zygotes. Yet, this relative simplicity does not preclude a conscientious preparatory work that is often neglected when initiating a project. Here, we describe the key steps leading to successful generation of a double knockout (KO) mouse by simultaneously targeting two homolog genes, Tmem176a and Tmem176b, which are located in the same genomic locus. Additionally, we show that similar efficiency can be obtained in a mixed genetic background or directly in the C57BL/6 inbred strain. Thus, presented as a detailed case study that should be helpful to the non-specialists, we focus on the genotyping strategy to anticipate the various possibilities.
Published: 2016
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22. Lineage Relationship Analysis of RORγt + Innate Lymphoid Cells

Author: Hans Jörg Fehling, Matthias Lochner, Naoko Satoh-Takayama, Marie Cherrier, James P. Di Santo, Francina Langa, Shinichiro Sawa, Gérard Eberl, Développement des Tissus Lymphoïdes, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Division of Experimental Virology, Centre for Experimental and Clinical Infection Research (TWINCORE), Helmholtz Centre for Infection Research (HZI)-Medizinische Hochschule Hannover (MHH)-Helmholtz Centre for Infection Research (HZI)-Medizinische Hochschule Hannover (MHH), Immunité Innée, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institute of Immunology [Ulm], Universitätsklinikum Ulm - University Hospital of Ulm, Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), This work was supported by the Institut Pasteur, grants from the Mairie de Paris, the Agence Nationale de la Recherche, and an Excellence Grant from the European Commission. M.L. was supported by the Deutsche Forschungsgemeinschaft and theSchlumberger Foundation. H.J.F. is funded by the Deutsche Forschungsgemeinschaft (FE-578/3-1)., Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), and Institut Pasteur [Paris]
Subjects: Receptors, CCR6, Adoptive cell transfer, Stromal cell, Lymphoid Tissue, Cellular differentiation, Mice, Transgenic, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Adaptive Immunity, Biology, Bacterial Physiological Phenomena, Proinflammatory cytokine, Mice, 03 medical and health sciences, Fetus, 0302 clinical medicine, T-Lymphocyte Subsets, RAR-related orphan receptor gamma, Intestine, Small, Animals, Homeostasis, Cell Lineage, Lymphocytes, Lymphopoiesis, Intestinal Mucosa, Symbiosis, Immunity, Mucosal, 030304 developmental biology, 0303 health sciences, Multidisciplinary, Innate lymphoid cell, Cell Differentiation, Lymphoid Progenitor Cells, Nuclear Receptor Subfamily 1, Group F, Member 3, Acquired immune system, Adoptive Transfer, Lymphocyte Subsets, Liver, Immunology, [SDV.IMM]Life Sciences [q-bio]/Immunology, 030215 immunology
Abstract: Innate Innit? Innate lymphocytes (ILCs) are a recently described population of immune cells that produce cytokines like those associated with T helper cells, but lack the recombined antigen receptors characteristic of T cells. Again, like some T helper cell lineages, a proportion of ILCs express the transcription factor RORγt. These include lymphoid tissue inducer (LTi) cells required for fetal lymphoid tissue organogenesis and a population of natural killer (NK)–like cells that function in gut immune responses. Sawa et al. (p. 665 ; see the Perspective by Veldhoen and Withers ) wondered whether the RORγt-expressing ILCs all develop from the same progenitor population. Indeed, they found a fetal liver progenitor that gave rise to several phenotypically distinct populations. However, the LTi cells were not progenitors for the NK-like cells. It seems the trajectory of different ILC populations is developmentally regulated, and postnatally ILCs are favored that play a role in intestinal defense before the gut is fully colonized by intestinal microbiota.
Published: 2010
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23. Characterization of the thymic IL-7 niche in vivo

Author: Ana Cumano, Odile Richard-Le Goff, Ana Patricia Sousa, Nuno L. Alves, Richard L. Boyd, Vera S. G. Ribeiro, Allison Bordack, Gérard Eberl, Lucie Peduto, James P. Di Santo, Ann P. Chidgey, Francina Langa Vives, Nicholas D. Huntington, Cytokines et Développement Lymphoïde, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Développement des Lymphocytes, Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), Développement des Tissus Lymphoïdes, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Monash Immunology and Stem Cell Laboratories (MISCL), Monash University [Clayton], Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris], and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Subjects: Genetically modified mouse, Chromosomes, Artificial, Bacterial, Chemokine, MESH: Chromosomes, Artificial, Bacterial, MESH: Mice, Transgenic, medicine.medical_treatment, T cell, Mice, Transgenic, Thymus Gland, Biology, Polymerase Chain Reaction, Mice, 03 medical and health sciences, 0302 clinical medicine, Genes, Reporter, In vivo, medicine, Animals, MESH: Animals, RNA, Messenger, MESH: Mice, Cells, Cultured, MESH: RNA, Messenger, 030304 developmental biology, 0303 health sciences, Thymic involution, Multidisciplinary, Interleukin-7, MESH: Genes, Reporter, CCL19, Epithelial Cells, MESH: Polymerase Chain Reaction, MESH: Thymus Gland, Biological Sciences, Molecular biology, MESH: Interleukin-7, Luminescent Proteins, Cytokine, medicine.anatomical_structure, MESH: Epithelial Cells, biology.protein, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Luminescent Proteins, CCL25, MESH: Cells, Cultured, 030215 immunology
Abstract: The thymus represents the “cradle” for T cell development, with thymic stroma providing multiple soluble and membrane cues to developing thymocytes. Although IL-7 is recognized as an essential factor for thymopoiesis, the “environmental niche” of thymic IL-7 activity remains poorly characterized in vivo. Using bacterial artificial chromosome transgenic mice in which YFP is under control of IL-7 promoter, we identify a subset of thymic epithelial cells (TECs) that co-express YFP and high levels of Il7 transcripts (IL-7 hi cells). IL-7 hi TECs arise during early fetal development, persist throughout life, and co-express homeostatic chemokines ( Ccl19 , Ccl25 , Cxcl12 ) and cytokines ( Il15 ) that are critical for normal thymopoiesis. In the adult thymus, IL-7 hi cells localize to the cortico-medullary junction and display traits of both cortical and medullary TECs. Interestingly, the frequency of IL-7 hi cells decreases with age, suggesting a mechanism for the age-related thymic involution that is associated with declining IL-7 levels. Our temporal-spatial analysis of IL-7-producing cells in the thymus in vivo suggests that thymic IL-7 levels are dynamically regulated under distinct physiological conditions. This IL-7 reporter mouse provides a valuable tool to further dissect the mechanisms that govern thymic IL-7 expression in vivo.
Published: 2009
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24. In vivo equilibrium of proinflammatory IL-17+ and regulatory IL-10+ Foxp3+ RORγt+ T cells

Author: Lucie Peduto, Marie Cherrier, Matthias Lochner, Francina Langa, Shinichiro Sawa, Mustapha Si-Tahar, James P. Di Santo, Gérard Eberl, Dieter Riethmacher, Rosa Varona, Hugot, Bérengère, Développement des Tissus Lymphoïdes, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), Department of Immunology and Oncology, Universidad Autónoma de Madrid (UAM)-Centro Nacional de Biotecnología [Madrid] (CNB-CSIC), Consejo Superior de Investigaciones Científicas [Madrid] (CSIC)-Consejo Superior de Investigaciones Científicas [Madrid] (CSIC), Development and Regeneration, Human Genetics Division, University of Southampton, Défense innée et inflammation, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Cytokines et Développement Lymphoïde, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], Universidad Autonoma de Madrid (UAM)-Centro Nacional de Biotecnologia, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris], and Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: MESH: Nuclear Receptor Subfamily 1, Group F, Member 3, MESH: Inflammation, MESH: Interleukin-10, Receptors, Retinoic Acid, MESH: Interleukin-17, T-Lymphocytes, MESH: Flow Cytometry, T-Lymphocytes, Regulatory, Mice, 0302 clinical medicine, MESH: Receptors, Interleukin-12, RAR-related orphan receptor gamma, Genes, Reporter, Immunology and Allergy, MESH: Animals, MESH: Genetic Variation, 0303 health sciences, education.field_of_study, Receptors, Thyroid Hormone, Interleukin-17, Receptors, Interleukin-12, FOXP3, hemic and immune systems, Forkhead Transcription Factors, Articles, Nuclear Receptor Subfamily 1, Group F, Member 3, Flow Cytometry, Interleukin-10, Interleukin 10, [SDV.IMM]Life Sciences [q-bio]/Immunology, Interleukin 17, medicine.medical_specialty, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH: Mice, Transgenic, Immunology, Population, chemical and pharmacologic phenomena, Mice, Transgenic, Biology, Article, Proinflammatory cytokine, 03 medical and health sciences, MESH: Forkhead Transcription Factors, Internal medicine, medicine, Animals, MESH: Receptors, Thyroid Hormone, education, MESH: Mice, 030304 developmental biology, MESH: Receptors, Retinoic Acid, Inflammation, Interleukin-6, MESH: T-Lymphocytes, Regulatory, T-cell receptor, MESH: Genes, Reporter, Genetic Variation, Molecular biology, MESH: Interleukin-6, CCL20, Endocrinology, MESH: T-Lymphocytes, 030215 immunology
Abstract: The nuclear hormone receptor retinoic acid receptor-related orphan receptor gamma t (RORgamma t) is required for the generation of T helper 17 cells expressing the proinflammatory cytokine interleukin (IL)-17. In vivo, however, less than half of RORgamma t(+) T cells express IL-17. We report here that RORgamma t(+) T alphabeta cells include Foxp3(+) cells that coexist with IL-17-producing RORgamma t(+) T alphabeta cells in all tissues examined. The Foxp3(+) RORgamma t(+) T alphabeta express IL-10 and CCL20, and function as regulatory T cells. Furthermore, the ratio of Foxp3(+) to IL-17-producing RORgamma t(+) T alphabeta cells remains remarkably constant in mice enduring infection and inflammation. This equilibrium is tuned in favor of IL-10 production by Foxp3 and CCL20, and in favor of IL-17 production by IL-6 and IL-23. In the lung and skin, the largest population of RORgamma t(+) T cells express the gammadelta T cell receptor and produce the highest levels of IL-17 independently of IL-6. Thus, potentially antagonistic proinflammatory IL-17-producing and regulatory Foxp3(+) RORgamma t(+) T cells coexist and are tightly controlled, suggesting that a perturbed equilibrium in RORgamma t(+) T cells might lead to decreased immunoreactivity or, in contrast, to pathological inflammation.
Published: 2008
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25. Conditional knock-out reveals that zygotic vezatin-null mouse embryos die at implantation

Author: Vincent Hyenne, Michel Cohen-Tannoudji, Francina Langa, Celine Souilhol, Christine Petit, Bernard Maro, Silvia Cereghini, Marie-Christine Simmler, Laboratoire de Biologie du Développement (LBD), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-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)-Centre National de la Recherche Scientifique (CNRS), Génétique Fonctionnelle de la Souris, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Génétique des Déficits Sensoriels, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Chaire Génétique et physiologie cellulaire, Collège de France (CdF (institution)), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], Sackler Faculty of Medicine, Tel Aviv University [Tel Aviv], Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), This work was supported by research Grants R0375/38 and R0475/100 from the Ligue Contre le Cancer (BM). Vincent Hyenne was recipient of a fellowship from the Ministère de l’Education et de la Recherche Technologique (MENRT) and from the Fondation pour la Recherche Médicale (FRM). Céline Souilhol is recipient of a fellowship from the Centre National de la Recherche Scientfique (CNRS). We thank Aude Jobart (IJM/CNRS UMR 7592) for her help with confocal microscopy. We are grateful to Rémi Beau and Françoise Thouron (Centre Ingéniérie Génétique Murine/Institut Pasteur) and Mélanie Fabre (Organogenèse Précoce chez la Souris et Maladies Génétiques Associées/CNRS UMR 7622) for excellent technical support., We sincerely thank Drs. Thierry Galli and Sophie Louvet-Vallée for critical review. We are grateful to Drs. Isabelle Roux, Yvan Lallemand and Shahragim Tajbakhsh for generous support and encouragement, 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)-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), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Collège de France - Chaire Génétique et physiologie cellulaire, Institut Pasteur [Paris] (IP), Tel Aviv University (TAU), and Cohen-Tannoudji, Michel
Subjects: Embryology, Mouse, Zygote, Pgk-1-cre, Peri-implantation lethality, Mutant, MESH: Amino Acid Sequence, MESH: Mice, Knockout, Mice, MESH: Embryo Implantation, Vezatin, 0302 clinical medicine, Embryonic Structure, MESH: Animals, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Mice, Knockout, 0303 health sciences, Embryo, Adherens Junctions, Cell biology, medicine.anatomical_structure, Essential gene, MESH: Membrane Proteins, Molecular Sequence Data, Intercellular adhesion, MESH: Carrier Proteins, Biology, Adherens junction, 03 medical and health sciences, [SDV.BDD] Life Sciences [q-bio]/Development Biology, medicine, Animals, Amino Acid Sequence, Embryo Implantation, Blastocyst, MESH: Mice, 030304 developmental biology, MESH: Molecular Sequence Data, MESH: Embryo, Mammalian, Membrane Proteins, Embryo, Mammalian, Embryonic stem cell, Molecular biology, Conditional vezatin knock-out, MESH: Adherens Junctions, Genes, Lethal, MESH: Zygote, MESH: Genes, Lethal, Carrier Proteins, 030217 neurology & neurosurgery, Developmental Biology
Abstract: International audience; Vezatin, a protein associated to adherens junctions in epithelial cells, is already expressed in mouse oocytes and during pre-implantation development. Using a floxed strategy to generate a vezatin-null allele, we show that the lack of zygotic vezatin is embryonic lethal, indicating that vezatin is an essential gene. Homozygous null embryos are able to elicit a decidual response but as early as day 6.0 post-coitum mutant implantation sites are devoid of embryonic structures. Mutant blastocysts are morphologically normal, but only half of them are able to hatch upon in vitro culture and the blastocyst outgrowths formed after 3.5 days in culture exhibit severe abnormalities, in particular disrupted intercellular adhesion and clear signs of cellular degeneration. Notably, the junctional proteins E-cadherin and beta-catenin are delocalized and not observed at the plasma membrane anymore. These in vitro observations reinforce the idea that homozygous vezatin-null mutants die at the time of implantation because of a defect in intercellular adhesion. Together these results indicate that the absence of zygotic vezatin is deleterious for the implantation process, most likely because cadherin-dependent intercellular adhesion is impaired in late blastocysts when the maternal vezatin is lost.
Published: 2007
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26. Protective role of LGP2 in influenza virus pathogenesis

Author: Pierre Charneau, Mustapha Si-Tahar, Viviane Balloy, Laurence Fiette, Michel Chignard, Julien Pothlichet, Damien Chopy, Monique Lafon, Fany Blanc, Laetitia Furio, Francina Langa, Défense innée et inflammation, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Histotechnologie et Pathologie, Institut Pasteur [Paris], Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Virologie moléculaire et Vectorologie, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], This work was supported by the Institut Pasteur (grant PTR 186), the Agence Nationale de la Recherche (grants FluProParand NKTVir to F. B. and L. F.), and the Ministère de l’Enseignement Supérieur et de la Recherche (fellowship to D. C.), ANR-07-MIME-0010,FLUPROPAR,Etude de la modulation de l'infection grippale par les protéases et les antiprotéases de l'hôte(2007), ANR-08-MIEN-0021,NKTVIR,Etude des mécanismes d'activation et rôle des lymphocytes T Natural Killer lors des infections virales(2008), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)
Subjects: MESH: Signal Transduction, [SDV]Life Sciences [q-bio], Gene Expression, medicine.disease_cause, [SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, DEAD-box RNA Helicases, Mice, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Leukocytes, Influenza A virus, Immunology and Allergy, MESH: Animals, MESH: Orthomyxoviridae Infections, 0303 health sciences, MESH: RNA Helicases, Pattern recognition receptor, 3. Good health, Infectious Diseases, MESH: Survival Analysis, Host-Pathogen Interactions, DEAD Box Protein 58, [SDV.IMM]Life Sciences [q-bio]/Immunology, Inflammation Mediators, Signal transduction, medicine.symptom, RNA Helicases, Signal Transduction, Genetically modified mouse, MESH: Gene Expression, MESH: Mice, Transgenic, Orthomyxoviridae, MESH: Inflammation Mediators, MESH: Influenza A virus, Mice, Transgenic, Inflammation, Biology, MESH: Leukocytes, 03 medical and health sciences, Immune system, Orthomyxoviridae Infections, In vivo, MESH: Mice, Inbred C57BL, medicine, Animals, MESH: DEAD-box RNA Helicases, MESH: Mice, 030304 developmental biology, MESH: Host-Pathogen Interactions, biology.organism_classification, Survival Analysis, Mice, Inbred C57BL, Disease Models, Animal, Immunology, MESH: Disease Models, Animal, 030215 immunology
Abstract: International audience; Influenza A virus triggers a contagious respiratory disease that can cause considerable morbidity and mortality. Using an in vitro approach, we previously demonstrated that the pattern recognition receptor retinoic acid-inducible gene I (RIG-I) plays a key role in influenza A virus-mediated immune response. However, the importance of RIG-I signaling in vivo has not been thoroughly examined, because of the lack of an appropriate mouse models. To circumvent this issue, we generated a new transgenic mouse overexpressing LGP2 (hereafter, "LGP2 TG mice"), a major regulator of the RIG-I signaling pathway. The time course of several parameters was compared in infected wild-type and LGP2 TG mice. We found that LGP2 TG mice displayed significantly reduced inflammatory mediators and a lower leukocyte infiltration into the bronchoalveolar airspace. More importantly, LGP2 TG mice had a significant survival advantage. Hence, our in vivo study reveals that LGP2 is a major downregulator of the influenza A virus-triggered detrimental inflammatory response.
Published: 2014
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27. Cellular and Behavioral Outcomes of Dorsal Striatonigral Neuron Ablation: New Insights into Striatal Functions

Author: Dorian Chabbert, Marianne Amalric, Hélène Marie, Pascal Salin, Lydia Kerkerian-Le Goff, Lena Concetta, Christophe Melon, Elisiana Tafi, Francina Langa, Corinne Beurrier, Delphine Révy, Florence Jaouen, Institut de Biologie du Développement de Marseille (IBDM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), European Brain Research Institute [Rome, Italy] (EBRI), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], Laboratoire de Neurosciences Cognitives [Marseille] (LNC), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Aix Marseille Université (AMU)-Collège de France (CdF (institution))-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)
Subjects: Dyskinesia, Drug-Induced, MESH: Dopamine Uptake Inhibitors, MESH: Antiparkinson Agents, Striatum, MESH: Movement, Anxiety, MESH: Corpus Striatum, Antiparkinson Agents, Levodopa, Cocaine, Dopamine Uptake Inhibitors, Basal ganglia, Neural Pathways, Direct pathway of movement, MESH: Animals, Diphtheria Toxin, Promoter Regions, Genetic, MESH: Levodopa, MESH: Diphtheria Toxin, MESH: Interneurons, Cholinergic Neurons, Substantia Nigra, Psychiatry and Mental health, medicine.anatomical_structure, MESH: Monosaccharide Transport Proteins, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Original Article, Heparin-binding EGF-like Growth Factor, MESH: Cholinergic Neurons, Monosaccharide Transport Proteins, MESH: Heparin-binding EGF-like Growth Factor, MESH: Mice, Transgenic, Movement, Neurotoxins, MESH: Substantia Nigra, Substantia nigra, Mice, Transgenic, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Nucleus accumbens, Biology, Medium spiny neuron, MESH: Cocaine, Interneurons, MESH: Promoter Regions, Genetic, medicine, Animals, Humans, Cholinergic neuron, MESH: Neurotoxins, Pharmacology, MESH: Humans, MESH: Dyskinesia, Drug-Induced, MESH: Anxiety, MESH: Neural Pathways, Ventral striatum, Corpus Striatum, nervous system, Neuroscience
Abstract: International audience; The striatum is the input structure of the basal ganglia network that contains heterogeneous neuronal populations, including two populations of projecting neurons called the medium spiny neurons (MSNs), and different types of interneurons. We developed a transgenic mouse model enabling inducible ablation of the striatonigral MSNs constituting the direct pathway by expressing the human diphtheria toxin (DT) receptor under the control of the Slc35d3 gene promoter, a gene enriched in striatonigral MSNs. DT injection into the striatum triggered selective elimination of the majority of striatonigral MSNs. DT-mediated ablation of striatonigral MSNs caused selective loss of cholinergic interneurons in the dorsal striatum but not in the ventral striatum (nucleus accumbens), suggesting a region-specific critical role of the direct pathway in striatal cholinergic neuron homeostasis. Mice with DT injection into the dorsal striatum showed altered basal and cocaine-induced locomotion and dramatic reduction of L-DOPA-induced dyskinesia in the parkinsonian condition. In addition, these mice exhibited reduced anxiety, revealing a role of the dorsal striatum in the modulation of behaviors involving an emotional component, behaviors generally associated with limbic structures. Altogether, these results highlight the implication of the direct striatonigral pathway in the regulation of heterogeneous functions from cell survival to regulation of motor and emotion-associated behaviors.
Published: 2014
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28. A Novel Nodal Enhancer Dependent on Pluripotency Factors and Smad2/3 Signaling Conditions a Regulatory Switch During Epiblast Maturation

Author: Anne Camus, Jérôme Collignon, Délara Sabéran-Djoneidi, Ataaillah Benhaddou, Alice Jouneau, Francina Langa, Valérie Mezger, Sascha Ott, Aitana Perea-Gomez, Costis Papanayotou, Université Paris Diderot, Sorbonne Paris Cité, Paris, France, Université Paris Diderot - Paris 7 (UPD7), Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Biologie du développement et reproduction (BDR), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Centre épigénétique et destin cellulaire (EDC), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), University College of London [London] (UCL), FRM, ANR, StemPole-IdF, Ile-de-France committees of Ligue Nationale contre le Cancer, Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Centre épigénétique et destin cellulaire (EDC (UMR_7216)), Institut Pasteur [Paris], Laboratory of Functional Genomics, Wolfson Institute for Biomedical Research, University College London, Wolfson Institute for Biomedical Research, and Centre National de la Recherche Scientifique (CNRS)-École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)
Subjects: Embryology, Cellular differentiation, [SDV]Life Sciences [q-bio], Nodal signaling, Smad2 Protein, souris, Cell Fate Determination, Mice, Cell Signaling, Animal Cells, Molecular Cell Biology, Biologie de la reproduction, Biology (General), Induced pluripotent stem cell, development, pluripotence, enhancer, epiblast, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Inhibin-beta Subunits, Genetics, Reproductive Biology, culture cellulaire, Stem Cells, General Neuroscience, Biologie du développement, Gene Expression Regulation, Developmental, virus diseases, Cell Differentiation, Nanog Homeobox Protein, Development Biology, épiblaste, 3. Good health, Cell biology, Enhancer Elements, Genetic, embryonic structures, Cellular Types, General Agricultural and Biological Sciences, Germ Layers, Research Article, Signal Transduction, Pluripotent Stem Cells, animal structures, Nodal Protein, QH301-705.5, DNA transcription, Mice, Transgenic, Germ layer, Biology, General Biochemistry, Genetics and Molecular Biology, Cell Line, cellule souche pluripotente, Molecular Genetics, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Smad3 Protein, Enhancer, QH426, Embryonic Stem Cells, Homeodomain Proteins, QL, différenciation cellulaire, [SDV.GEN]Life Sciences [q-bio]/Genetics, Biology and life sciences, General Immunology and Microbiology, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Cell Biology, Molecular Development, QP, Embryonic stem cell, Gene regulation, [SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics, [SDV.BDD.EO]Life Sciences [q-bio]/Development Biology/Embryology and Organogenesis, Epiblast, embryon, Gene expression, NODAL, Octamer Transcription Factor-3, Developmental Biology
Abstract: HBE, a newly discovered enhancer element, mediates the influence of pluripotency factors and Activin/Nodal signaling on early Nodal expression in the mouse embryo, and controls the activation of later-acting Nodal enhancers., During early development, modulations in the expression of Nodal, a TGFβ family member, determine the specification of embryonic and extra-embryonic cell identities. Nodal has been extensively studied in the mouse, but aspects of its early expression remain unaccounted for. We identified a conserved hotspot for the binding of pluripotency factors at the Nodal locus and called this sequence “highly bound element” (HBE). Luciferase-based assays, the analysis of fluorescent HBE reporter transgenes, and a conditional mutation of HBE allowed us to establish that HBE behaves as an enhancer, is activated ahead of other Nodal enhancers in the epiblast, and is essential to Nodal expression in embryonic stem cells (ESCs) and in the mouse embryo. We also showed that HBE enhancer activity is critically dependent on its interaction with the pluripotency factor Oct4 and on Activin/Nodal signaling. Use of an in vitro model of epiblast maturation, relying on the differentiation of ESCs into epiblast stem cells (EpiSCs), revealed that this process entails a shift in the regulation of Nodal expression from an HBE-driven phase to an ASE-driven phase, ASE being another autoregulatory Nodal enhancer. Deletion of HBE in ESCs or in EpiSCs allowed us to show that HBE, although not necessary for Nodal expression in EpiSCs, is required in differentiating ESCs to activate the differentiation-promoting ASE and therefore controls this regulatory shift. Our findings clarify how early Nodal expression is regulated and suggest how this regulation can promote the specification of extra-embryonic precusors without inducing premature differentiation of epiblast cells. More generally, they open new perspectives on how pluripotency factors achieve their function., Author Summary In the early mouse embryo, Nodal, a member of the TGFbeta superfamily of signalling proteins, promotes the differentiation of extra-embryonic tissues, as well as tissues within the developing embryo itself. Characterising the regulation of Nodal gene expression is essential to understand how Nodal signals in diverse tissue types and at different stages of embryonic development. Four distinct enhancer sequences have been shown to regulate Nodal expression, although none could account for it in the preimplantation epiblast or in embryonic stem cells. We identified a novel enhancer, HBE, responsible for the earliest aspects of Nodal expression. We show that activation of HBE depends on its interaction with a well-known pluripotency factor called Oct4. HBE itself also controls the activation of at least one other Nodal enhancer. Our findings clarify how early Nodal expression is regulated and reveal how pluripotency factors may control the onset of differentiation in embryonic tissues.
Published: 2014
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29. Lineage tracing and genetic ablation of ADAM12(+) perivascular cells identify a major source of profibrotic cells during acute tissue injury

Author: Francina Langa, Lucie Peduto, Gérard Eberl, Sophie Dulauroy, Selene Di Carlo, Développement des Tissus Lymphoïdes, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP), This work was funded by the Institut Pasteur, grants from the Mairie de Paris, the Agence Nationale de la Recherche and an Excellence grant from the European Commission. S.E.D.C. is funded by La Ligue contre le Cancer, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris]
Subjects: MESH: Leg Injuries, Pathology, Receptor, Platelet-Derived Growth Factor alpha, medicine.medical_treatment, MESH: Receptor, Platelet-Derived Growth Factor alpha, [SDV]Life Sciences [q-bio], Freund's Adjuvant, ADAM12 Protein, Cobra Cardiotoxin Proteins, MESH: Ear, External, Mice, 0302 clinical medicine, Fibrosis, Genes, Reporter, MESH: Collagen, Adipocytes, MESH: Freund's Adjuvant, MESH: Animals, Ear, External, Myofibroblasts, 0303 health sciences, MESH: Muscle, Skeletal, General Medicine, Dermis, MESH: ADAM12 Protein, MESH: Crosses, Genetic, Cell biology, Specific Pathogen-Free Organisms, MESH: Cobra Cardiotoxin Proteins, medicine.anatomical_structure, 030220 oncology & carcinogenesis, Gene Knockdown Techniques, Acute Disease, MESH: Fibrosis, MESH: Acute Disease, [SDV.IMM]Life Sciences [q-bio]/Immunology, Collagen, MESH: Parabiosis, MESH: ADAM Proteins, medicine.medical_specialty, Stromal cell, MESH: Myofibroblasts, MESH: Mice, Transgenic, Parabiosis, Mice, Transgenic, Biology, General Biochemistry, Genetics and Molecular Biology, Proinflammatory cytokine, 03 medical and health sciences, Cicatrix, Fate mapping, medicine, Animals, Cell Lineage, Progenitor cell, Muscle, Skeletal, MESH: Mice, Crosses, Genetic, MESH: Adipocytes, 030304 developmental biology, Wound Healing, MESH: Cicatrix, Growth factor, MESH: Genes, Reporter, MESH: Blood Vessels, MESH: Cell Lineage, medicine.disease, MESH: Dermis, MESH: Gene Knockdown Techniques, MESH: Specific Pathogen-Free Organisms, ADAM Proteins, MESH: Wound Healing, Blood Vessels, Stromal Cells, MESH: Stromal Cells, Wound healing, Leg Injuries
Abstract: International audience; Profibrotic cells that develop upon injury generate permanent scar tissue and impair organ recovery, though their origin and fate are unclear. Here we show that transient expression of ADAM12 (a disintegrin and metalloprotease 12) identifies a distinct proinflammatory subset of platelet-derived growth factor receptor-α-positive stromal cells that are activated upon acute injury in the muscle and dermis. By inducible genetic fate mapping, we demonstrate in vivo that injury-induced ADAM12(+) cells are specific progenitors of a major fraction of collagen-overproducing cells generated during scarring, which are progressively eliminated during healing. Genetic ablation of ADAM12(+) cells, or knockdown of ADAM12, is sufficient to limit generation of profibrotic cells and interstitial collagen accumulation. ADAM12(+) cells induced upon injury are developmentally distinct from muscle and skin lineage cells and are derived from fetal ADAM12(+) cells programmed during vascular wall development. Thus, our data identify injury-activated profibrotic progenitors residing in the perivascular space that can be targeted through ADAM12 to limit tissue scarring.
Published: 2012
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30. I-SceI-mediated double-strand break does not increase the frequency of homologous recombination at the Dct locus in mouse embryonic stem cells.: Homologous recombination at the Dct locus

Author: Fenina, Myriam, Simon-Chazottes, Dominique, Vandormael-Pournin, Sandrine, Soueid, Jihane, Langa, Francina, Cohen-Tannoudji, Michel, Bernard, Bruno A, Panthier, Jean-Jacques, Génétique Fonctionnelle de la Souris, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), L'OREAL, Research & Innovation, Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], This work was supported by grants from the Association Nationale de la Recherche et de la Technologie (ANRT), research contract C080052, and L’Oréal Recherche. MF was awarded fellowships from the ANRT, L’Oréal Recherche and Fondation pour la Recherche Médicale. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript., Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)
Subjects: MESH: DNA Damage, MESH: DNA Primers, [SDV.GEN]Life Sciences [q-bio]/Genetics, MESH: Alleles, [SDV]Life Sciences [q-bio], MESH: Polymerase Chain Reaction, MESH: Base Sequence, MESH: Gene Targeting, MESH: Intramolecular Oxidoreductases, body regions, MESH: Homologous Recombination, MESH: Animals, MESH: Embryonic Stem Cells, MESH: Mice, MESH: Deoxyribonucleases, Type II Site-Specific
Abstract: International audience; Targeted induction of double-strand breaks (DSBs) at natural endogenous loci was shown to increase the rate of gene replacement by homologous recombination in mouse embryonic stem cells. The gene encoding dopachrome tautomerase (Dct) is specifically expressed in melanocytes and their precursors. To construct a genetic tool allowing the replacement of Dct gene by any gene of interest, we generated an embryonic stem cell line carrying the recognition site for the yeast I-SceI meganuclease embedded in the Dct genomic segment. The embryonic stem cell line was electroporated with an I-SceI expression plasmid, and a template for the DSB-repair process that carried sequence homologies to the Dct target. The I-SceI meganuclease was indeed able to introduce a DSB at the Dct locus in live embryonic stem cells. However, the level of gene targeting was not improved by the DSB induction, indicating a limited capacity of I-SceI to mediate homologous recombination at the Dct locus. These data suggest that homologous recombination by meganuclease-induced DSB may be locus dependent in mammalian cells.
Published: 2012
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31. Loss of central and peripheral CD8+ T-cell tolerance to HFE in mouse models of human familial hemochromatosis

Author: Boucherma, Rachid, Kridane-Miledi, Hédia, Vives, Francina Langa, Vauchy, Charline, Borg, Christophe, Kleinclauss, François, Fiette, Laurence, Tiberghien, Pierre, Lemonnier, François, Rohrlich, Pierre, Huetz, François, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté]), Centre d'Investigation Clinique de Besançon (Inserm CIC 1431), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté]), Immunité Cellulaire Antivirale, Institut Pasteur [Paris], Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Service d'Oncologie Médicale [CHRU Besançon], Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Service d'urologie, andrologie et transplantation rénale, Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Hôpital Saint-Jacques, Histopathologie humaine et Modèles animaux, Département de pédiatrie, Saas, Philippe, Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Institut Pasteur [Paris] (IP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Université de Franche-Comté (UFC), and Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Université de Franche-Comté (UFC)
Subjects: MESH: Hemochromatosis, MESH: Genetic Diseases, Inborn, MESH: Immune Tolerance, [SDV.IMM] Life Sciences [q-bio]/Immunology, Mutation, Missense, Graft vs Host Disease, MESH: Graft vs Host Disease, Thymus Gland, CD8-Positive T-Lymphocytes, MESH: Mice, Knockout, MESH: Histocompatibility Antigens Class I, Mice, MESH: Skin, Immune Tolerance, MESH: Transplantation, Homologous, Animals, Humans, Transplantation, Homologous, MESH: Animals, Hemochromatosis Protein, MESH: Mice, Skin, Mice, Knockout, MESH: Mutation, Missense, MESH: Humans, MESH: Mice, Inbred DBA, Histocompatibility Antigens Class I, Genetic Diseases, Inborn, Membrane Proteins, MESH: Thymus Gland, MESH: Amino Acid Substitution, MESH: CD8-Positive T-Lymphocytes, Disease Models, Animal, Amino Acid Substitution, Mice, Inbred DBA, [SDV.IMM]Life Sciences [q-bio]/Immunology, Hemochromatosis, MESH: Membrane Proteins, MESH: Disease Models, Animal
Abstract: International audience; HFE, an MHC class Ib molecule that controls iron metabolism, can be directly targeted by cytotoxic TCR αβ T lymphocytes. Transgenic DBA/2 mice expressing, in a Rag 2 KO context, an αβ TCR that directly recognizes mouse HFE (mHFE) were created to further explore the interface of HFE with the immune system. TCR-transgenic mHfe WT mice deleted mHFE-reactive T cells in the thymus, but a fraction of reprogrammed cells were able to escape deletion. In contrast, TCR-transgenic mice deprived of mHFE molecules (mHfe KO mice) or expressing a C282→Y mutated mHFE molecule - the most frequent mutation associated with human hereditary hemochromatosis - positively selected mHFE-reactive CD8(+) T lymphocytes and were not tolerant toward mHFE. By engrafting these mice with DBA/2 WT (mHFE(+)) skin, it was established, as suspected on the basis of similar engraftments performed on DBA/2 mHfe KO mice, that mHFE behaves as an autonomous skin-associated histocompatibility antigen, even for mHFE-C282→Y mutated mice. By contrast, infusion of DBA/2 mHFE(+) mice with naïve mHFE-reactive transgenic CD8(+) T lymphocytes did not induce GVHD. Thus, tolerance toward HFE in mHfe WT mice can be acquired at either thymic or peripheral levels but is disrupted in mice reproducing human familial hemochromatosis.
Published: 2012
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32. Field evaluation of rapid diagnostic tests for meningococcal meningitis in Niger

Author: Pascal Boisier, Suzanne Chanteau, Saacou Djibo, Farida Nato, A. Amadou Hamidou, A. Elhaj Mahamane, Fati Sidikou, Centre de Recherche Médicale et Sanitaire (Niamey, Niger) (CERMES), Réseau International des Instituts Pasteur (RIIP), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris], Institut Pasteur de Nouvelle-Calédonie, and Institut Pasteur [Paris] (IP)
Subjects: medicine.medical_specialty, Time Factors, Concordance, 030231 tropical medicine, Reference laboratory, Meningitis, Meningococcal, Neisseria meningitidis, medicine.disease_cause, Polymerase Chain Reaction, Sensitivity and Specificity, MESH: Neisseria meningitidis, 03 medical and health sciences, 0302 clinical medicine, Internal medicine, parasitic diseases, medicine, MESH: Reagent Strips, Humans, 030212 general & internal medicine, Serotyping, health care economics and organizations, Reagent Strips, Rapid diagnostic test, Antigens, Bacterial, MESH: Humans, business.industry, MESH: Time Factors, Public Health, Environmental and Occupational Health, MESH: Meningitis, Meningococcal, Diagnostic test, MESH: Serotyping, MESH: Polymerase Chain Reaction, Dipstick, medicine.disease, equipment and supplies, MESH: Sensitivity and Specificity, 3. Good health, Surgery, Infectious Diseases, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Acute Disease, Meningococcal meningitis, MESH: Acute Disease, Parasitology, business, Meningitis, MESH: Antigens, Bacterial
Abstract: Summary Objective To evaluate dipstick rapid diagnostic tests (RDTs) for meningococcal meningitis in basic health facilities. Methods Health facility staff received a one-day training. During the meningitis season, they performed RDTs on cerebrospinal fluid (CSF) specimens from suspected cases of meningitis. A frozen aliquot of CSF was later tested using polymerase chain reaction (PCR) to establish the reference diagnosis. RDTs used in health facilities were archived to allow checking the concordance between reported diagnosis and observed results. Reported diagnosis was also compared to PCR diagnosis. A second RDT was performed on each CSF specimen at the reference laboratory. Results Using RDTs, health facilities reported 382 negative results (73.9%), 114 NmA (22.1%), 12 NmW135 (2.3%) and nine uninterpretable results (1.7%), the latter corresponding to the misuse of a reagent by three agents. The agreement between reported diagnosis and archived dipsticks was excellent (kappa = 0.98). The agreement between PCR diagnosis and reported RDTs results was strong (kappa = 0.82). In health facilities, the sensitivity of RDTs for N. meningitidis A was Se = 0.91. The kappa coefficient measuring the agreement between RDTs operated in the reference laboratory and RDTs operated in health facilities was κ = 0.78. Conclusion We confirmed that dipstick RDTs to identify N. meningitidis serogroups A, C, W135 and Y can be reliably operated by non-specialized staff in basic health facilities. RDTs proved very useful to recommend vaccination in NmA epidemics, and also to avoid vaccination in epidemics due to serogroups not included in vaccines (NmX).
Published: 2008
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32 results on '"Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM)"'

1. Mice Humanized for MHC and hACE2 with High Permissiveness to SARS-CoV-2 Omicron Replication

2. Clonal hematopoiesis driven by chromosome 1q/MDM4 trisomy defines a canonical route toward leukemia in Fanconi anemia

3. SPICE‐Met : profiling and imaging energy metabolism at the single‐cell level using a fluorescent reporter mouse

4. Rab35-regulated lipid turnover by myotubularins represses mTORC1 activity and controls myelin growth

5. An intranasal lentiviral booster reinforces the waning mRNA vaccine-induced SARS-CoV-2 immunity that it targets to lung mucosa

6. De novo germline mutation in the dual specificity phosphatase 10 gene accelerates autoimmune diabetes

7. Brain Cross‐Protection against SARS‐CoV‐2 Variants by a Lentiviral Vaccine in New Transgenic Mice

8. The antitumoral effects of chemerin are independent from leukocyte recruitment and mediated by inhibition of neoangiogenesis

9. Dynamics of myogenic differentiation using a novel Myogenin knock-in reporter mouse

10. Dissection of the Fgf8 regulatory landscape by in vivo CRISPR-editing reveals extensive intra- and inter-enhancer redundancy

11. Lysosomal Acid Lipase Drives Adipocyte Cholesterol Homeostasis and Modulates Lipid Storage in Obesity, Independent of Autophagy

12. Corrigendum: Expression of Bioactive Chemerin by Keratinocytes Inhibits Late Stages of Tumor Development in a Chemical Model of Skin Carcinogenesis

13. Mitochondrial AIF Loss Causes Metabolic Reprogramming, Caspase-Independent Cell Death Blockade, Embryonic Lethality, and Perinatal Hydrocephalus

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

15. A distinct cardiopharyngeal mesoderm genetic hierarchy establishes antero-posterior patterning of esophagus striated muscle

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

17. A Human Polymorphism in CHRNA5 Is Linked to Relapse to Nicotine Seeking in Transgenic Rats

18. Reduced recruitment of 53BP1 during interstrand crosslink repair is associated with genetically inherited attenuation of mitomycin C sensitivity in a family with Fanconi anemia

19. Penicillin Resistance Compromises Nod1-Dependent Proinflammatory Activity and Virulence Fitness of Neisseria meningitidis

20. MITF – A controls branching morphogenesis and nephron endowment

21. Generation of a Double KO Mouse by Simultaneous Targeting of the Neighboring Genes Tmem176a and Tmem176b Using CRISPR/Cas9: Key Steps from Design to Genotyping

22. Lineage Relationship Analysis of RORγt + Innate Lymphoid Cells

23. Characterization of the thymic IL-7 niche in vivo

24. In vivo equilibrium of proinflammatory IL-17+ and regulatory IL-10+ Foxp3+ RORγt+ T cells

25. Conditional knock-out reveals that zygotic vezatin-null mouse embryos die at implantation

26. Protective role of LGP2 in influenza virus pathogenesis

27. Cellular and Behavioral Outcomes of Dorsal Striatonigral Neuron Ablation: New Insights into Striatal Functions

28. A Novel Nodal Enhancer Dependent on Pluripotency Factors and Smad2/3 Signaling Conditions a Regulatory Switch During Epiblast Maturation

29. Lineage tracing and genetic ablation of ADAM12(+) perivascular cells identify a major source of profibrotic cells during acute tissue injury

30. I-SceI-mediated double-strand break does not increase the frequency of homologous recombination at the Dct locus in mouse embryonic stem cells.: Homologous recombination at the Dct locus

31. Loss of central and peripheral CD8+ T-cell tolerance to HFE in mouse models of human familial hemochromatosis

32. Field evaluation of rapid diagnostic tests for meningococcal meningitis in Niger

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32 results on '"Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM)"'

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