Characterization of a host hepatocyte factor that regulates HCV replication

描述(由申请人提供)：慢性丙型肝炎病毒(丙型肝炎病毒)感染发生在所有暴露的个人中的70%-80%，影响世界人口的3%。需要更有效和耐受性更好的丙型肝炎病毒治疗方法。我们的长期目标是定义宿主-丙型肝炎病毒的分子相互作用，并通过这样做，为丙型肝炎病毒治疗寻找新的治疗剂。我们已经使用全基因组siRNA文库完成了用于丙型肝炎病毒复制的宿主辅助因子的功能性基因组筛选。在众多新的宿主辅助因子中，PI4KA是一种磷脂酰肌醇4-激酶(PI-4-Kinase)。我们假设PI4KA是形成丙型肝炎病毒复制所需的改变的宿主膜所必需的。本提案描述了一项为期5年的指导性培训计划，目的是在分子病毒学、肌醇磷脂和细胞贩运以及细胞成像领域获得进一步的技术技能。这些技术将被用于开发新的模型系统，以研究丙型肝炎病毒复制复合体及其相关的宿主膜和蛋白质的形成和组成。本项目的具体目的是(1)确定PI4KA抑制丙型肝炎病毒复制的特异性，(2)确定PI4KA在病毒复制周期中的作用，以及(3)开发系统来研究丙型肝炎病毒复制复合体的组装和组成。虽然具体目标是针对PI4KA，但在获奖期间将开发的系统和技术将应用于丙型肝炎病毒复制的其他宿主辅助因子的研究。这项拟议的研究将直接对公众健康产生好处，因为更好地了解强调丙型肝炎病毒复制复合体组装的机制可能会为丙型肝炎治疗带来新的靶点。更好地了解丙型肝炎病毒与宿主的关系是《美国国立卫生研究院肝病研究行动计划》(目标B2a)的明确目标。我们最近对丙型肝炎病毒复制所需的人类(宿主)蛋白进行了全基因组筛查。这些蛋白质是丙型肝炎病毒新疗法的潜在靶点。这项建议描述了一项为期5年的研究计划，重点是开发新的系统，以研究宿主辅助因子如何指导丙型肝炎病毒复制复合体的组装。

DESCRIPTION (provided by applicant): Chronic hepatitis C virus (HCV) infection develops in 70-80% of all exposed individuals and affects 3% of the world's population. There is a need for more effective and better-tolerated HCV therapies. Our long-term goals are to define host-HCV molecular interactions and in so doing, to identify novel therapeutic agents for HCV therapy. We have completed a functional genomic screen for host cofactors for HCV replication using a whole- genome siRNA library. Among the many novel host cofactors identified is PI4KA, a phosphatidylinositol 4- kinase (PI 4-kinase). We hypothesize that PI4KA is necessary for formation of the altered host membranes required for HCV replication. This proposal describes a 5 year mentored training program to acquire further technical skills in the fields of molecular virology, phosphoinositides and cell trafficking, and cell imaging. These skills will be used to develop novel model systems to study the formation and composition of the HCV replication complex and its associated host membranes and proteins. The specific aims of this project are (1) to characterize the specificity of PI4KA inhibition for suppression of HCV replication, (2) to characterize the role of PI4KA through the viral replication cycle, and (3) to develop systems to study HCV replication complex assembly and composition. Although the specific aims are directed towards PI4KA, the systems and techniques to be developed during the award period will be applied to the study of other host cofactors of HCV replication. The proposed research will have direct public health benefits as a better understanding of the mechanisms that underline HCV replication complex assembly may lead to novel targets for HCV therapies. A better understanding of the HCV-host relationship is a stated goal of the NIH Action Plan for Liver Disease Research (Goal B2a). We have recently performed a whole-genome screen for human (host) proteins that are required by the hepatitis C virus for its replication. Such proteins are potential targets for new therapies for HCV. This proposal describes a 5-year research plan focusing on developing new systems to study how host cofactors direct the assembly of HCV replication complexes.