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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：
8658799
负责人：
Charles M Rice
金额：
$ 54.67万
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/8658799
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 biophysical techniques 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（由申请人提供）：这是2006年首次资助的“参与HCV进入的细胞因子的鉴定和表征”的竞争性更新。本实验室对丙型肝炎病毒（HCV）基因表达和复制机制的研究已有20多年的历史，但由于研究体系不完善，对病毒进入机制的理解受到限制。我们和其他人通过开发第一个研究HCV体外进入的平台- HCV假颗粒（HCVpp）和完全感染性HCV细胞培养系统（HCVpp）克服了这一障碍。随着这些新工具的出现，我们提出了识别和表征HCV摄取所需的因素。这些研究的一个长期目标是利用这些信息来建立急需的HCV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HCV进入因子。我们通过阐明病毒进入小鼠细胞所需的人为因素来建立这一发现，现在已经建立了第一个支持整个HCV生命周期的基因人源化小鼠模型。这一成就为研究HCV进入体内并利用小鼠遗传学的力量对其进行解剖提供了前所未有的机会。我们还在被动和主动疫苗策略中证明了这种动物模型的概念。除了推进体内模型，我们正在寻求更多的生物相关的细胞培养系统的病毒。这些包括原代成人和胎儿肝细胞培养物和三维肝脏类器官。在这种竞争性的更新中，我们采用我们的新的体外和体内系统，以进一步了解HCV进入所需的病毒-宿主相互作用。我们提出了遗传和生物物理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，探测病毒与宿主相关脂质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。