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（HCV）感染发生在所有暴露个体的70-80%中，影响世界人口的3%。需要更有效和耐受性更好的HCV疗法。我们的长期目标是确定宿主-HCV分子相互作用，并在此过程中，确定用于HCV治疗的新型治疗药物。我们已经完成了一个功能基因组筛选宿主辅因子的HCV复制使用全基因组siRNA文库。在许多新的宿主辅因子中鉴定出的是PI 4KA，一种磷脂酰肌醇4-激酶（PI 4-激酶）。我们假设PI 4KA是形成HCV复制所需的宿主膜改变所必需的。该提案描述了一个为期5年的指导培训计划，以获得分子病毒学，磷酸肌醇和细胞贩运以及细胞成像领域的进一步技术技能。这些技能将用于开发新的模型系统，以研究HCV复制复合物及其相关宿主膜和蛋白质的形成和组成。该项目的具体目的是（1）表征PI 4KA抑制对HCV复制的抑制的特异性，（2）表征PI 4KA在病毒复制周期中的作用，以及（3）开发研究HCV复制复合物组装和组成的系统。虽然具体目标是针对PI 4KA，但在奖励期间开发的系统和技术将应用于HCV复制的其他宿主辅因子的研究。拟议的研究将有直接的公共卫生利益，因为更好地了解HCV复制复合物组装的机制可能会导致HCV治疗的新靶点。更好地了解HCV-宿主关系是NIH肝病研究行动计划的既定目标（目标B2 a）。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选。这些蛋白质是HCV新疗法的潜在靶点。该提案描述了一项为期5年的研究计划，重点是开发新系统，研究宿主辅因子如何指导HCV复制复合物的组装。

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.