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Identification and characterization of cellular factors involved in HCV entry

参与 HCV 进入的细胞因子的鉴定和表征

基本信息

批准号：
8463102
负责人：
Charles M Rice
金额：
$ 51.39万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-12-15 至 2017-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8463102
关键词：
Achievement Adult Animal Model Apolipoproteins Applications Grants Binding CD81 gene Cell Culture System Cell surface Cells Cirrhosis Coxsackie B Viruses Cytoskeleton Engineering Event Fibrosis Funding Gene Expression Genetic Goals Grant Growth HCV Animal Models Hand Hepatitis C Hepatitis C virus Hepatocyte Human Immunocompetent Immunology In Vitro Individual Infection Infectious hepatitides Integration Host Factors Kinetics Laboratories Life Cycle Stages Light Lipids Lipoproteins Liver Low Density Lipoprotein Receptor Maps Mediating Metabolism Modeling Molecular Biology Monitor Mus Nature Organoids Parasites Pathogenesis Pathway interactions Plasmodium falciparum Play Population Preparation Primary carcinoma of the liver cells Process Proteins Proteomics Protocols documentation Reporting Resolution Role SR-BI receptor Scheme Signal Transduction Structure Subfamily lentivirinae System Testing Tight Junctions Time Vaccines Variant Viral Virion Virus Virus Diseases Virus Receptors Work base claudin-1 protein drug testing fetal improved in vivo in vivo Model inhibitor/antagonist insight laser tweezer lipid metabolism monolayer mouse model novel occludin particle pathogen receptor tool uptake vaccine evaluation virus culture virus host interaction

项目摘要

DESCRIPTION (provided by applicant): This is a competing renewal of a grant "Identification and characterization of cellular factors involved in HCV entry" first funded in 2006. Our laboratory has been studying the mechanisms of hepatitis C virus (HCV) gene expression and replication for over twenty years, but our understanding of viral entry was limited by inadequate study systems. We and others overcame this hurdle by developing the first platforms for studying HCV entry in vitro - HCV pseudoparticles (HCVpp) and the fully-infectious HCV cell culture system (HCVcc). With the advent of these new tools, we proposed to identify and characterize factors required for HCV uptake. A long- term goal of these studies was to use this information to create much needed small animal models for HCV. Using lentivirus-based cyclic repackaging schemes, we succeeded in identifying two tight junction proteins, claudin-1 and occludin, as critical HCV entry factors. We built on this finding by elucidating the human factors required for viral entry into mouse cells, and have now gone on to create the first genetically humanized mouse model supporting the entire HCV lifecycle. This achievement opens unprecedented opportunities to study HCV entry in vivo and to dissect it using the power of mouse genetics. We have also shown proof-of-concept for this animal model in passive and active vaccine strategies. In addition to advancing in vivo models, we are pursuing more biologically relevant cell culture systems for the virus. These include primary adult and fetal hepatocyte cultures and three-dimensional liver organoids. In this competing renewal, we employ our novel in vitro and in vivo systems to further understand the virus-host interactions required for HCV entry. We propose genetic and biophysical approaches to uncover the mechanisms of uptake. Our studies will, for the first time, define the composition and structure of the infecting particle, probe the interplay between virus and host- associated lipid metabolism factors, and map the molecular interaction network active during viral internalization.

描述(由申请者提供)：这是2006年首次资助的“识别和表征进入丙型肝炎病毒所涉及的细胞因素”资助的竞争性续期。本实验室对丙型肝炎病毒基因表达和复制机制的研究已有二十多年的历史，但由于研究系统的不完善，我们对病毒进入的认识受到限制。我们和其他人克服了这一障碍，开发了研究丙型肝炎病毒体外进入的第一个平台-丙型肝炎病毒假颗粒(HCVpp)和完全感染性丙型肝炎病毒细胞培养系统(HCVcc)。随着这些新工具的出现，我们建议识别和表征丙型肝炎病毒感染所需的因素。这些研究的长期目标是利用这些信息来创建急需的丙型肝炎病毒小动物模型。使用基于慢病毒的循环重新包装方案，我们成功地确定了两个紧密连接蛋白，claudin-1和occludin，作为关键的丙型肝炎病毒进入因子。在这一发现的基础上，我们阐明了病毒进入小鼠细胞所需的人类因素，现在已经创造了第一个支持整个丙型肝炎病毒生命周期的遗传人化小鼠模型。这一成就为研究丙型肝炎病毒在体内的进入并利用小鼠遗传学的力量剖析它打开了前所未有的机会。我们还展示了被动和主动疫苗策略对该动物模型的概念验证。除了推进体内模型外，我们还在为病毒寻求更具生物学意义的细胞培养系统。这些包括成人和胎儿肝细胞的原代培养和三维肝脏器官培养。在这一竞争性更新中，我们使用我们的新型体外和体内系统来进一步了解丙型肝炎病毒进入所需的病毒-宿主相互作用。我们建议用遗传学和生物物理学的方法来揭示摄取的机制。我们的研究将首次确定感染颗粒的组成和结构，探索病毒和宿主相关脂代谢因子之间的相互作用，并绘制病毒内化过程中活跃的分子相互作用网络。