Modeling GBV-B Replication in Immortalized Cell Lines

性状（由申请方提供）：尚无可复制丙型肝炎病毒（HCV）的稳健细胞培养系统。因此，HCV复制的替代模型经常用于评价抗病毒化合物。GB病毒B（GBV-B）是与HCV在遗传学上最密切相关的病毒，是HCV药物开发工作的有吸引力的替代模型。不幸的是，GBV-B迄今只能在取自小型非人灵长类动物的原代肝细胞中生长。这使得目前的GBV-B抗病毒测定难以进行，昂贵，并且不适合高通量（HTP）筛选。本申请旨在利用永生化细胞系开发GBV-B复制的稳健细胞培养模型。该项目的具体目标是使GBV-B易感动物的原代肝细胞永生化，鉴定这些细胞系和其他已建立的细胞系是否可以支持GBV-B体外复制，并标准化和验证用于评价抗病毒药物的模型系统。该项目将使GBV-B抗病毒检测广泛可用，负担得起，并适合HTP应用。这将有助于发现和开发新的抗HCV药物，从而改善HCV感染患者的生活质量。

DESCRIPTION (provided by applicant): There is no robust cell culture system in which the hepatitis C virus (HCV) can replicate. As such, surrogate models of HCV replication are frequently used for the evaluation of antiviral compounds. The GB virus B (GBV-B) is the closest related virus phylogenetically with HCV and is an attractive surrogate model for HCV drug development efforts. Unfortunately, GBV-B can so far only be grown in primary hepatocytes taken from small non-human primates. This makes the current GBV-B antiviral assay difficult to perform, expensive, and not amenable to high-throughput (HTP) screening. This application aims to develop a robust cell culture model of GBV-B replication utilizing immortalized cell lines. The specific aims of this project are to immortalize the primary hepatocytes of GBV-B-susceptible animals, identify if these and other established cell lines can support GBV-B replication in vitro, and to standardize and validate the model system produced for use in the evaluation of antiviral agents. This project will make the GBV-B antiviral assay widely available, affordable, and appropriate for HTP applications. This will facilitate the discovery and development of new anti-HCV drugs that may improve the quality of life for HCV-infected patients.