Contributors: Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL); Institut Pasteur de Lille; Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire CHU Lille (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS); Copenhagen Hepatitis C Program Copenhague, Danemark (CO-HEP); Hvidovre Hospital-University of Copenhagen = Københavns Universitet (UCPH); Unité de Virologie clinique et fondamentale (UVCF); Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie; Récepteurs nucléaires, maladies cardiovasculaires et diabète - U 1011 (RNMCD); Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire CHU Lille (CHRU Lille); Centre de recherche sur l'Inflammation (CRI (UMR_S_1149 / ERL_8252 / U1149)); Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS); Service d’Hépatologie Hôpital Beaujon; Hôpital Beaujon AP-HP; Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP); Institut de Recherche sur les Maladies Virales et Hépatiques (IVH); Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM); Institut de biologie de Lille - UMS 3702 (IBL); Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Université de Lille-Centre National de la Recherche Scientifique (CNRS); This work was supported by the French National Agency for Research on AIDS and Viral Hepatitis (ANRS) and the ANR through the ERA-NET Infect-ERA program (grant ANR-13-IFEC-0002-01). In addition, this study was supported by research grants from the Danish Council for Independent Research-Medical Sciences and the Novo Nordisk Foundation. The BioImaging Center Lille Nord-de-France is supported by the Equipex IMAGINEX program.; The Copenhagen Hepatitis C Program (CO-HEP) is a joint venture of the Department of Infectious Diseases and Clinical Research Centre, Hvidovre Hospital, and the Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark. We thank F. L. Cosset, C. Rice, and T. Wakita for providing essential reagents. We also thank S. Ung for his help in preparing the figures and L. Linna for manuscript proofreading. We are very grateful to A. Danneels for her precious help during the revisions. Immunofluorescence and flow cytometry analyses were performed with the help of the imaging core facility of the BioImaging Center Lille Nord-de-France.; ANR-10-EQPX-0004,Imaginex BioMed,Plateau de microscopie de criblage à haut débit et d'analyse à très haute résolution(2010); ANR-13-IFEC-0002,HCV-ASSEMBLY,Identification of host factors involved in Hepatitis C Virus assembly and characterization of their potential role in vivo(2013)
نبذة مختصرة : International audience ; Hepatitis C virus (HCV) infection causes 500,000 deaths annually, in association with end-stage liver diseases. Investigations of the HCV life cycle have widened the knowledge of virology, and here we discovered that two piperazinylbenzenesulfonamides inhibit HCV entry into liver cells. The entry of HCV into host cells is a complex process that is not fully understood but is characterized by multiple spatially and temporally regulated steps involving several known host factors. Through a high-content virus infection screening analysis with a library of 1,120 biologically active chemical compounds, we identified SB258585, an antagonist of serotonin receptor 6 (5-HT6), as a new inhibitor of HCV entry in liver-derived cell lines as well as primary hepatocytes. A functional characterization suggested a role for this compound and the compound SB399885, which share similar structures, as inhibitors of a late HCV entry step, modulating the localization of the coreceptor tight junction protein claudin-1 (CLDN1) in a 5-HT6-independent manner. Both chemical compounds induced an intracellular accumulation of CLDN1, reflecting export impairment. This regulation correlated with the modulation of protein kinase A (PKA) activity. The PKA inhibitor H89 fully reproduced these phenotypes. Furthermore, PKA activation resulted in increased CLDN1 accumulation at the cell surface. Interestingly, an increase of CLDN1 recycling did not correlate with an increased interaction with CD81 or HCV entry. These findings reinforce the hypothesis of a common pathway, shared by several viruses, which involves G-protein-coupled receptor-dependent signaling in late steps of viral entry.
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