Role of Lipids in Hepatitis C Virus Maturation

描述（由申请人提供）：慢性丙型肝炎病毒感染影响全球约1.7亿人，是纤维化、脂肪变性和肝细胞癌（HCC）的重要风险因素。丙型肝炎病毒感染是肝移植的主要指标。在这里，我们提出的特点的作用，脂质在病毒形态发生主要集中在成熟和分泌从感染的肝细胞。在以前的资助中，我们已经确定并表征了几种脂质和脂质相互作用蛋白在影响HCV分泌中的作用。 在这里，我们建议继续这些研究，并专注于磷脂酰肌醇-4磷酸（PI 4P）相互作用的蛋白质，包括神经酰胺转移蛋白（CERT），Nir-2，和Arfaptin在影响跨高尔基体网络的HCV病毒体的分泌运输的作用。HCV通过高尔基体网络的成熟/分泌被认为与极低密度脂蛋白（VLDL）组装和分泌相关。VLDL转运发生在专门的VLDL转运囊泡（VTV）中。我们建议使用一个已建立的生化分离程序隔离VTV跨高尔基体网络的HCV相关VTV的运输的特点。在含有HCV病毒体成分的HCV感染细胞中，VTV的表征对于理解HCV形态发生具有根本重要性。我们还建议，以确定功能的重要性，影响HCV成熟的VTV的因素。这些研究将揭示对HCV成熟、分泌和排出机制的独特见解，并将作为其他RNA病毒的模型。本研究的结果将为影响HCV分泌途径的细胞靶点的治疗设计开辟新的途径。

DESCRIPTION (provided by applicant): Chronic Hepatitis C virus infections affect about 170 million people worldwide and constitute a significant risk factor for fibrosis, steatosis and hepatocellular carcinoma (HCC). HCV infections are a major indicator for liver transplantation. Here, we propose to characterize the role of lipids in virion morphogenesis mostly focusing on maturation and secretion from infected hepatocytes. In the previous grant we had identified and characterized the role of several lipids and lipid interacting proteins in affecting HCV secretion. Here, we propose to continue these studies and focus on the role of phosphatidylinosito-4 phosphate (PI4P)-interacting proteins including ceramide transfer protein (CERT), Nir-2, and Arfaptin in affecting the secretory transport of HCV virions across the Golgi network. HCV maturation/secretions through the Golgi network is believed to occur in association with very low-density lipoprotein (VLDL) assembly and secretion. VLDL transport occurs in specialized VLDL transport vesicles (VTVs). We propose to characterize the transport of HCV-associated VTVs across the Golgi network using an established biochemical fractionation procedure for isolating VTVs. Characterization of VTVs in HCV infected cells containing HCV virions components is of fundamental importance to the understating of HCV morphogenesis. We also propose to determine the functional importance of factors that affect VTVs in HCV maturation. These studies will reveal unique insight into the mechanisms of HCV maturation, secretion and egress and will serve as a model for other RNA viruses. The results of this study will open new avenues for therapeutic design of cellular targets affecting the HCV secretory pathway.