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长期感染了美国约400万人，在全球范围内感染了1.7亿人。 HCV与HIV的共同感染非常普遍，总体25-30％的HIV阳性患者，尤其是在共同感染70％的毒品滥用者中（2）。 HCV感染是HCC开发的主要危险因素。肝病相关的HCV相关末期是肝移植的主要指示。由于缺乏强大的细胞培养HCV传播系统以及可靠的HCV感染和复制的小型动物模型，HCV研究的进步受到了显着阻碍。最近的突破是开发可靠的细胞培养系统，用于复制HCV复制子RNA和感染性HCV的产生，这允许对整个HCV生命周期的遗传研究。但是，HCV感染和复制的小动物模型的发展是具有挑战性的，缺乏可靠的小动物模型代表了HCV研究的主要障碍。最近，我们证明了基因型2a HCV RNA在小鼠胚胎成纤维细胞（MEF）中有效复制，尽管在小鼠肝细胞中效率低下。更重要的是，我们已经证明了细胞内的cDNA衍生的HCV RNA导致感染性HCV的强大产生。查理·赖斯（Charlie Rice）小组最近的一项研究表明，小鼠肝细胞中人CD81和闭合蛋白的表达是必不可少的，足以感染HCV假型颗粒。这些显着的进步为开发新型HCV感染和复制的新型转基因小鼠模型提供了坚定的基础。我们假设细胞类型特异性细胞蛋白对于有效的HCV RNA复制很重要，并且在肝细胞中表达人CD81和occludin的转基因小鼠易受到HCV感染的影响。该探索性研究建议的总体目标是开发新型的转基因HCV小鼠模型。具体而言，我们将使用蛋白质组学，生化和遗传互补方法确定对小鼠细胞中有效HCV RNA复制重要的细胞蛋白（特定目标1）； 2）开发新的转基因小鼠，该小鼠要么包含表达荧光素酶的JFH1 HCV RNA的全长cDNA，要么在肝特异性启动子的控制下表达人CD81和occludin（特定的目标2）； 3）与小鼠干扰素和HCV NS3蛋白酶 - 和NS5B聚合酶特异性抑制剂的施用结合使用荧光素酶的HCV，确定转基因小鼠的HCV感染和复制（特定的AIM 3）。转基因HCV小鼠模型的成功发展将代表HCV研究领域的范式转移，并使我们能够确定病毒和细胞蛋白在HCV感染，复制，发病机理以及癌变以及非法药物和酒精对HCVENISES的影响HCV和酒精的作用。此外，转基因HCV小鼠模型将促进抗HCV药物发现和HCV疫苗发育，最终控制HCV感染。 公共卫生相关性：丙型肝炎病毒（HCV）感染仍然是一个主要的全球健康问题，影响了全球约1.7亿人和400万美国人。我们的研究建议的总体目标是开发新型的转基因HCV小鼠模型，以确定HCV复制，发病机理，致癌作用以及非法药物和酒精对HCV复制和发病机理的影响。此外，转基因HCV小鼠模型将促进抗病毒药物发现和疫苗发育，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