Role of Lipids in Hepatitis C Virus Maturation

脂质在丙型肝炎病毒成熟中的作用

• 批准号: 8286215
• 负责人: ALEEM SIDDIQUI
• 金额: $ 38.24万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8286215
• 关键词: 1,2-diacylglycerol; Actomyosin; Affect; Affinity; Antiviral Agents; Biochemical; Cell-Free System; Cells; Ceramides; Chronic Hepatitis; Cirrhosis; Complex; Density Gradient Centrifugation; Diglycerides; Fatty Liver; Fractionation; Genome; Glycoproteins; Goals; Golgi Apparatus; Hepatitis C; Hepatitis C virus; Immunofluorescence Immunologic; Infection; Integration Host Factors; Internal Ribosome Entry Site; Investigation; Laser Microscopy; Lipids; Liver diseases; Location; Maintenance; Malignant neoplasm of liver; Membrane Protein Traffic; Minority; Modeling; Morphogenesis; Nucleocapsid; Pathway interactions; Patients; Phosphatidylinositols; Phosphorylation; Phosphotransferases; Play; Population; Primary carcinoma of the liver cells; Procedures; Process; Proteins; Proteomics; Public Health; RNA; RNA Viruses; RNA replication; Regulation; Ribonucleoproteins; Role; Rough endoplasmic reticulum; Secretory Vesicles; Shapes; Stretching; Structure; Sucrose; Translating; Transport Vesicles; Very low density lipoprotein; Vesicle; Viral; Virion; Work; base; design; inhibitor/antagonist; inorganic phosphate; insight; interest; iodixanol; liver transplantation; microsomal triglyceride transfer protein; novel; oxysterol binding protein; particle; protein function; protein kinase D; public health relevance; secretion process; trafficking; viral RNA

DESCRIPTION (provided by applicant): About 3% of world population is infected with Hepatitis C virus (HCV). HCV infection often leads to chronic hepatitis and about one third of patients develop cirrhosis and a minority of those develops hepatocellular carcinoma. The infection is also associated with fatty liver disease. HCV replicates its RNA genome within ribonucleoprotein complexes (RNP) consisting of viral and host cellular factors. One such host factor, termed oxysterol binding protein (OSBP) was identified by an affinity based approach by us. Subsequent studies revealed that OSBP was necessary for HCV secretion/release with no obvious effect on intracellular RNA replication. It belongs to group of proteins referred to as phosphatidyinosito-4 phosphate (PI4P)-interacting proteins. In this application, we focus on the investigation of OSBP and two other PI4P-interacting proteins, CERT and GOLPH3 in HCV maturation/secretion pathway. Role of Golgi kinase PKD in HCV maturation will also investigated. HCV maturation/secretion, which likely occurs in the Golgi compartment, is widely believed to occur in association with very low density lipoprotein (VLDL) particles. We propose to characterize the HCV maturation process through Golgi compartments by confocal laser microscopy and immunofluorescence in the context of OSBP and VLDL secretory pathway. VLDL particles are secreted via specialized VLDL transport vesicles (VTVs). Using an established biochemical fractionation procedure of isolation of VTVs, we propose to determine the association of HCV components with VTVs. Characterization of VTVs in HCV infected cells containing HCV virion components is of fundamental importance in understanding the HCV morphogenesis. Identification of host factors involved in this assembly/secretion process will also open new avenues for designing novel antiviral strategies. These studies will reveal unique insights into the mechanisms of HCV maturation, secretion and budding processes and provide a unique model for other RNA viruses. PUBLIC HEALTH RELEVANCE: HCV infections progresses to cirrhosis, steatosis, and liver cancer and are the leading indications for liver transplants thus representing a major public health burden. About 3% of world population is infected with HCV.

描述（由申请人提供）：世界人口中约 3% 感染丙型肝炎病毒 (HCV)。 HCV 感染通常会导致慢性肝炎，约三分之一的患者会发展为肝硬化，其中少数会发展为肝细胞癌。这种感染还与脂肪肝疾病有关。 HCV 在由病毒和宿主细胞因子组成的核糖核蛋白复合物 (RNP) 内复制其 RNA 基因组。我们通过基于亲和力的方法鉴定了一种这样的宿主因子，称为氧甾醇结合蛋白（OSBP）。随后的研究表明，OSBP是HCV分泌/释放所必需的，对细胞内RNA复制没有明显影响。它属于称为磷脂酰肌醇 4 磷酸 (PI4P) 相互作用蛋白的蛋白质组。在此应用中，我们重点研究 HCV 成熟/分泌途径中的 OSBP 和其他两种 PI4P 相互作用蛋白 CERT 和 GOLPH3。还将研究高尔基体激酶 PKD 在 HCV 成熟中的作用。 HCV 成熟/分泌可能发生在高尔基体中，人们普遍认为与极低密度脂蛋白 (VLDL) 颗粒有关。我们建议在 OSBP 和 VLDL 分泌途径的背景下，通过共聚焦激光显微镜和免疫荧光来表征 HCV 通过高尔基室的成熟过程。 VLDL 颗粒通过专门的 VLDL 转运囊泡 (VTV) 分泌。使用已建立的 VTV 分离生化分馏程序，我们建议确定 HCV 成分与 VTV 的关联。含有 HCV 病毒颗粒成分的 HCV 感染细胞中 VTV 的表征对于理解 HCV 形态发生至关重要。识别这种组装/分泌过程中涉及的宿主因子也将为设计新型抗病毒策略开辟新途径。这些研究将揭示对HCV成熟、分泌和出芽过程机制的独特见解，并为其他RNA病毒提供独特的模型。 公共卫生相关性：HCV 感染会进展为肝硬化、脂肪变性和肝癌，是肝移植的主要适应症，因此构成了重大的公共卫生负担。世界人口中约 3% 感染 HCV。