Transgenic mouse models of HCV infection and replication

HCV感染和复制的转基因小鼠模型

• 批准号: 8204439
• 负责人: Jieyun Jiang
• 金额: $ 22.28万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-03 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8204439
• 关键词: Affect; Albumins; Alcohols; American; Animal Model; Antiviral Agents; Biochemical Genetics; CD81 gene; Catalytic RNA; Cell Culture System; Cell Culture Techniques; Cell Line; Cells; Chronic Hepatitis; Cirrhosis; Complementary DNA; DNA Polymerase II; Development; Embryo; Fibroblasts; Foundations; Genetic; Genetic Transcription; Genome; Genomics; Genotype; Goals; HCV Vaccine; HIV; HIV Seropositivity; Hepatitis C; Hepatitis C virus; Hepatitis Delta Virus; Hepatocyte; Human; Illicit Drugs; Infection; Infectious hepatitides; Interferons; Knowledge; Laboratories; Length; Life Cycle Stages; Liver; Liver diseases; Luciferases; Measures; Molecular; Mouse Cell Line; Mus; Pathogenesis; Peptide Hydrolases; Persons; Polymerase; Primary carcinoma of the liver cells; Production; Proteins; Proteomics; RNA; RNA Sequences; RNA replication; Replicon; Research; Research Proposals; Rice; Risk Factors; Role; Staging; Structure; System; Transgenic Mice; Transgenic Organisms; Variant; Viral; Viral Proteins; Virion; Virus; Virus Receptors; Virus Replication; Work; anti-hepatitis C; base; carcinogenesis; cell type; drug abuser; drug development; drug discovery; global health; hepatitis C virus NS3 protein; hepatoma cell; in vivo; inhibitor/antagonist; innovation; interdisciplinary approach; liver transplantation; mouse model; novel; occludin; particle; promoter; public health relevance; receptor; vaccine development; viral RNA

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) is the leading cause of chronic hepatitis, cirrhosis, and hepatocellular carcinoma (HCC). HCV chronically infects approximately 4 million people in the U.S. and 170 million people worldwide. HCV co-infection with HIV is very common with overall 25-30% of HIV-positive persons, particularly among drug abusers with up to 70% co-infection (2). HCV infection is a major risk factor for HCC development. HCV- associated end-stage of liver diseases is the leading indication for liver transplantation. Advances on HCV research have been significantly hampered by the lack of a robust cell culture HCV propagation system and reliable small animal models of HCV infection and replication. Recent breakthroughs have been the development of robust cell culture systems for replication of HCV replicon RNAs and production of infectious HCV, which allows genetic studies of the entire HCV life cycle. However, development of small animal models of HCV infection and replication is challenging and that the lack of a reliable small animal model represents a major barrier to HCV research. Recently, we have demonstrated that genotype 2a HCV RNA replicated efficiently in mouse embryonic fibroblasts (MEF) albeit inefficiently in mouse hepatocytes. More importantly, we have demonstrated that the cDNA-derived HCV RNA inside the cell resulted in robust production of infectious HCV. A recent study by Charlie Rice's group has demonstrated that the expression of human CD81 and occludin in mouse hepatocytes is essential and sufficient for infection of HCV pseudotyped particles. These remarkable advances provide a firm foundation to develop novel transgenic mouse models of HCV infection and replication. We hypothesize that cell type-specific cellular proteins are important for efficient HCV RNA replication and that transgenic mice expressing human CD81 and occludin in hepatocytes are susceptible to HCV infection. The overall goal of this exploratory research proposal is to develop novel transgenic HCV mouse models. Specifically, we will 1) identify cellular proteins important for efficient HCV RNA replication in mouse cells using proteomics, biochemical, and genetic complementation approaches (Specific Aim 1); 2) develop novel transgenic mice that either contain full-length cDNAs of luciferase-expressing JFH1 HCV RNAs or express both human CD81 and occludin under the control of a liver-specific promoter (Specific Aim 2); and 3) determine HCV infection and replication in transgenic mice using luciferase-expressing HCV in conjunction with the administration of mouse interferon and HCV NS3 protease- and NS5B polymerase-specific inhibitors (Specific Aim 3). The successful development of transgenic HCV mouse models will represent a paradigm shift in the field of HCV research and will allow us to determine the roles and mechanisms of action of viral and cellular proteins in HCV infection, replication, pathogenesis, and carcinogenesis as well as the effects of illicit drugs and alcohol on HCV replication and pathogenesis. Additionally, transgenic HCV mouse models will facilitate the anti-HCV drug discovery and HCV vaccine development to eventually control HCV infection. PUBLIC HEALTH RELEVANCE: Hepatitis C virus (HCV) infection remains a major global health problem, affecting approximately 170 million people worldwide and 4 million Americans. Our overall goal of this research proposal is to develop novel transgenic HCV mouse models for the determination of the molecular mechanisms of HCV replication, pathogenesis, carcinogenesis, and the effects of illicit drugs and alcohol on HCV replication and pathogenesis. Additionally, transgenic HCV mouse models will facilitate the antiviral drug discovery and vaccine development to eventually control HCV infection.

描述（由申请人提供）：丙型肝炎病毒（HCV）是慢性肝炎、肝硬化和肝细胞癌（HCC）的主要原因。HCV慢性感染美国约400万人，全球约1.7亿人。HCV与HIV合并感染非常常见，总体上有25-30%的HIV阳性者，特别是在药物滥用者中，合并感染率高达70%（2）。HCV感染是HCC发展的主要危险因素。HCV相关的终末期肝病是肝移植的主要适应症。HCV研究的进展一直受到缺乏一个强大的细胞培养HCV繁殖系统和可靠的小动物模型的HCV感染和复制显着阻碍。最近的突破是开发了用于复制HCV复制子RNA和产生感染性HCV的稳健的细胞培养系统，这允许对整个HCV生命周期进行遗传研究。然而，HCV感染和复制的小动物模型的开发是具有挑战性的，并且缺乏可靠的小动物模型是HCV研究的主要障碍。最近，我们已经证明，基因型2a HCV RNA复制有效地在小鼠胚胎成纤维细胞（MEF），虽然效率低下，在小鼠肝细胞。更重要的是，我们已经证明了细胞内的cDNA衍生的HCV RNA导致了感染性HCV的稳健产生。Charlie Rice小组最近的一项研究表明，小鼠肝细胞中人CD 81和闭合蛋白的表达对于HCV假型颗粒的感染是必要的和足够的。这些显著的进展为开发新型HCV感染和复制的转基因小鼠模型提供了坚实的基础。我们假设细胞类型特异性细胞蛋白对于HCV RNA的有效复制是重要的，并且在肝细胞中表达人CD 81和occludin的转基因小鼠对HCV感染易感。这项探索性研究计划的总体目标是开发新型转基因HCV小鼠模型。具体来说，我们将1）确定细胞蛋白质的重要性，有效的丙型肝炎病毒RNA复制在小鼠细胞中使用蛋白质组学，生物化学和遗传互补的方法（具体目标1）; 2）开发新的转基因小鼠，其含有表达JFH 1 HCV RNA的全长cDNA或在肝脏特异性启动子的控制下表达人CD 81和闭合蛋白（具体目的2）;和3）使用表达HCV的转基因小鼠，结合施用小鼠干扰素和HCV NS 3蛋白酶-和NS 5 B聚合酶-特异性抑制剂，测定HCV感染和复制（具体目的3）。转基因HCV小鼠模型的成功开发将代表HCV研究领域的范式转变，并将使我们能够确定病毒和细胞蛋白在HCV感染，复制，发病机制和致癌作用以及非法药物和酒精对HCV复制和发病机制的影响。此外，转基因HCV小鼠模型将有助于抗HCV药物的发现和HCV疫苗的开发，以最终控制HCV感染。 公共卫生关系：丙型肝炎病毒（HCV）感染仍然是一个主要的全球健康问题，影响全球约1.7亿人和400万美国人。我们这项研究的总体目标是开发新的转基因HCV小鼠模型，用于确定HCV复制，发病机制，致癌作用的分子机制，以及非法药物和酒精对HCV复制和发病机制的影响。此外，转基因HCV小鼠模型将有助于抗病毒药物的发现和疫苗的开发，以最终控制HCV感染。

项目成果

Recent advance in antiviral drugs for hepatitis C.

• DOI: 10.3969/j.issn.1672-7347.2011.11.001
• 发表时间: 2011-11-01
• 期刊: Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences
• 作者: Liu, Jia;Shi, Shuang;Luo, Guangxiang
• 通讯作者: Luo, Guangxiang