title: In vitro efficacy of approved and experimental antivirals against novel genotype 3 hepatitis C virus subgenomic replicons.
abstract: Infection with genotype 3 hepatitis C virus (HCV) is common throughout the world, however no direct-acting antiviral (DAA) has been approved to treat this genotype. We therefore attempted to develop novel genotype 3 replicons to facilitate the discovery and development of new HCV therapies. A novel Huh-7-derived cell line 1C but not Lunet cells enabled the selection of a few stable colonies of a genotype 3a subgenomic replicon (strain S52). Genotypic analysis revealed a mutation of P89L in the viral NS3 protease domain, which was confirmed to enhance genotype 3a RNA replication and enable the establishment of highly replicating luciferase-encoding replicons. Secondary adaptive mutations that further enhanced RNA replication were identified in the viral NS3 and NS4A proteins. In addition, cell lines that were cured of genotype 3a replicons demonstrated higher permissiveness specifically to genotype 3a HCV replication. These novel replicons and cell lines were then used to study the activity of approved and experimental HCV inhibitors. NS3 protease and non-nucleoside NS5B polymerase inhibitors often demonstrated substantially less antiviral activity against genotype 3a compared to genotype 1b. In contrast, nucleoside analog NS5B inhibitors and host-targeting HCV inhibitors showed comparable antiviral activity between genotypes 3a and 1b. Overall, the establishment of this novel genotype 3a replicon system, in conjunction with those derived from other genotypes, will aid the development of treatment regimens for all genotypes of HCV.
authors.affiliation: Biology Department, Gilead Sciences, 333 Lakeside Drive, Foster City, CA 94404, USA.
authors.forename: Mei
authors.initials: M
authors.lastname: Yu
authors.forename: Amoreena C
authors.initials: AC
authors.lastname: Corsa
authors.forename: Simin
authors.initials: S
authors.lastname: Xu
authors.forename: Betty
authors.initials: B
authors.lastname: Peng
authors.forename: Ruoyu
authors.initials: R
authors.lastname: Gong
authors.forename: Yu-Jen
authors.initials: YJ
authors.lastname: Lee
authors.forename: Katie
authors.initials: K
authors.lastname: Chan
authors.forename: Hongmei
authors.initials: H
authors.lastname: Mo
authors.forename: William
authors.initials: W
authors.lastname: Delaney
authors.forename: Guofeng
authors.initials: G
authors.lastname: Cheng
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D017930:Genes, Reporter; D005838:Genotype; D016174:Hepacivirus / Q000145:classification / Q000187:drug effects* / Q000235:genetics; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D008826:Microbial Sensitivity Tests / Q000379:methods; D012093:Replicon / Q000187:drug effects*; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D008156:Luciferases
keywords: 2-C-methyl adenosine; 2-CMeA; 50% effective inhibitory concentration; CsA; DAA; Direct-acting antivirals; EC(50); G418; GT; Genotype 3 HCV; HCV; Huh-7 1C cells; NS; P89L mutation in NS3; Pan-genotype; RLU; Renilla luciferase; Rluc; cyclosporin A; direct-acting antivirals; geneticin; genotype; hepatic C virus; neo; neomycin transferase gene; non-structural; relative light units