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Hepatitis C virus: Morphogenesis, infection and therapy

World Journal of Hepatology 2018年第2期10卷 186-212页

作者： Vladimir Alexei Morozov sylvie lagaye Center for HIV and Retrovirology Department of Infectious Diseases Robert Koch Institute Department of Immunology Institut PasteurINSERM

Hepatitis C virus(HCV) is a major cause of liver diseases including liver cirrhosis and hepatocellular carcinoma. Approximately 3% of the world population is infected with HCV. Thus, HCV infection is considered a public healthy challenge. It is worth mentioning, that the HCV prevalence is dependent on the countries with infection rates around 20% in high endemic countries. The review summarizes recent data on HCV molecular biology, the physiopathology of infection(immune-mediated liver damage, liver fibrosis and lipid metabolism), virus diagnostic and treatment. In addition, currently available in vitro, ex vivo and animal models to study the virus life cycle, virus pathogenesis and therapy are described. Understanding of both host and viral factors may in the future lead to creation of new approaches in generation of an efficient therapeutic vaccine.

关键词： Hepatitis C virus Transmission Molecular biology Pathogenesis In vitro and ex vivo models of hepatitis C virus infection Treatment

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Adult human liver slice cultures:Modelling of liver fibrosis and evaluation of new anti-fibrotic drugs

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World Journal of Hepatology 2021年第2期13卷 187-217页

作者： Daria Kartasheva-Ebertz Jesintha Gaston Loriane Lair-Mehiri Pierre-Philippe Massault Olivier Scatton Jean-Christophe Vaillant Vladimir Alexei Morozov Stanislas Pol sylvie lagaye Institut Pasteur Immunobiologie des Cellules DendritiquesINSERM U1223Paris 75015France BioSPC Universitéde ParisParis 75005France Service de Chirurgie digestive Hépato-biliaire et EndocrinienneAPHPGroupe Hospitalier CochinParis 75014France Service de Chirurgie digestive et Hépato bilio pancréatique AP-HPGroupe Hospitalier La Pitié-SalpétrièreMedecine Sorbonne UniversitéParis 75013France Center for HIV and Retrovirology Department of Infectious DiseasesRobert Koch InstituteBerlin 13353Germany Département d'Hépatologie AP-HPGroupe Hospitalier CochinUniversitéde ParisParis 75014France

BACKGROUND Liver fibrosis can result in end-stage liver failure and death.AIM To examine human liver fibrogenesis and anti-fibrotic therapies,we evaluated the three dimensional ex vivo liver slice(LS)model.METHODS Fibrotic liver samples(F0 to F4 fibrosis stage according to the METAVIR score)were collected from patients after liver resection.Human liver slices(HLS)were cultivated for up to 21 days.Hepatitis C virus(HCV)infection,alcohol(ethanol stimulation)and steatosis(palmitate stimulation)were examined in fibrotic(F2 to F4)liver slices infected(or not)with HCV.F0-F1 HLS were used as controls.At day 0,either ursodeoxycholic acid(choleretic and hepatoprotective properties)and/or α-tocopherol(antioxidant properties)were added to standard of care on HLS and fibrotic liver slices,infected(or not)with HCV.Expression of the biomarkers of fibrosis and the triglyceride production were checked by quantitative reverse transcription polymerase chain reaction and/or enzymelinked immunosorbent assay.RESULTS The cultures were viable in vitro for 21 days allowing to study fibrosis inducers and to estimate the effect of anti-fibrotic drugs.Expression of the biomarkers of fibrosis and the progression to steatosis(estimated by triglycerides production)was increased with the addition of HCV and/or ethanol or palmitate.From day 15 of the follow-up studies,a significant decrease of both transforming growth factorβ-1 and Procol1A1 expression and triglycerides production was observed when a combined anti-fibrotic treatment was applied on HCV infected F2-F4 LS cultures.CONCLUSION These results show that the human three dimensional ex vivo model effectively reflects the in vivo processes in damaged human liver(viral,alcoholic,nonalcoholic steatohepatitis liver diseases)and provides the proof of concept that the LS examined model permits a rapid evaluation of new anti-fibrotic therapies when used alone or in combination.

关键词： Human liver fibrosis Hepatitis C virus Alcoholic liver disease Nonalcoholic steatohepatitis Ex vivo model Drugs

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Anti-hepatitis C virus potency of a new autophagy inhibitor using human liver slices model

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World Journal of Hepatology 2016年第21期8卷 902-914页

作者： sylvie lagaye Sonia Brun Jesintha Gaston Hong Shen Ruzena Stranska Claire Camus Clarisse Dubray Géraldine Rousseau Pierre-Philippe Massault Jerome Courcambeck Firas Bassisi Philippe Halfon Stanislas Pol Institut Pasteur INSERM Institut Cochin INSERM U1016CNRS UMR8104Universite Paris Descartes(UMR S1016) Genoscience Pharma KU Leuven Rega Institute Laboratoire Alphabio APHP Groupe Hospitalier La Pitie Salpe-triereService de Chirurgie digestive et Hepato bilio pancreatique APHP Groupe Hospitalier CochinService de Chirurgie digestiveHepato-biliaire et Endocrinienne Universite Paris Descartes APHP Groupe Hospitalier CochinUnite d’Hepatologie Institut Pasteur Departement de Recherche TranslationnelleINSERM

AIM: To evaluate the antiviral potency of a new antihepatitis C virus(HCV) antiviral agent targeting the cellular autophagy machinery. METHODS: Non-infected liver slices, obtained from human liver resection and cut in 350 μm-thick slices(2.7 × 106 cells per slice) were infected with cell culture-grown HCV Con1b/C3 supernatant(multiplicity of infection = 0.1) cultivated for up to ten days. HCV infected slices were treated at day 4 post-infection with GNS-396 for 6 d at different concentrations. HCV replication was evaluated by strand-specific real-time quantitative reverse transcription- polymerase chain reaction. The infectivity titers of supernatants were evaluated by foci formation upon inoculation into naive Huh-7.5.1 cells. The cytotoxic effect of the drugs was evaluated by lactate dehydrogenase leakage assays. RESULTS: The antiviral efficacy of a new antiviral drug, GNS-396, an autophagy inhibitor, on HCV infection of adult human liver slices was evidenced in a dosedependent manner. At day 6 post-treatment, GNS-396 EC50 was 158 nmol/L without cytotoxic effect(compared to hydroxychloroquine EC50 = 1.17 μmol/L).CONCLUSION: Our results demonstrated that our ex vivo model is efficient for evaluation the potency of autophagy inhibitors, in particular a new quinoline derivative GNS-396 as antiviral could inhibit HCV infection in a dosedependent manner without cytotoxic effect.

关键词： Host antiviral therapy Hepatitis C virus Tissue culture Autophagy Quinoline derivative

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