Characterization and Consequences of HCV Interaction with Host Lipoproteins

HCV 与宿主脂蛋白相互作用的特征和后果

• 批准号: 8298947
• 负责人: Margaret Adele Scull
• 金额: $ 5.22万
• 依托单位: ROCKEFELLER UNIVERSITY
• 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 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8298947
• 关键词: Address; Antibodies; Binding; Biochemical; Biological Assay; CD46 Antigen; Cataloging; Catalogs; Cell Culture Techniques; Cells; Cholesterol; Chronic; Complement; Complement Activation; Complex; Cytolysis; Data; Deposition; Development; Disease; Epitopes; Exhibits; Failure; Glycoproteins; Goals; HIV; Hepatitis C; Hepatitis C Prevalence; Hepatitis C virus; Hepatocyte; Heterogeneity; High Density Lipoproteins; Host Defense Mechanism; Humoral Immunities; Immune system; In Vitro; Individual; Infection; Infection Control; Kinetics; Lead; Licensing; Life Cycle Stages; Link; Lipids; Lipoproteins; Liver Cirrhosis; Mass Spectrum Analysis; Mediating; Membrane Glycoproteins; Molecular; Mutagenesis; Nature; Outcome; Pathway interactions; Patients; Population; Population Heterogeneity; Predisposition; Primary carcinoma of the liver cells; Proteins; Proteomics; Role; SR-BI receptor; Satellite Viruses; Serum; System; Techniques; Technology; Testing; United States; Vaccines; Very low density lipoprotein; Viral; Virion; Virus; Virus Diseases; Work; base; density; design; effective therapy; in vivo; influenzavirus; insight; liver transplantation; neutralizing antibody; particle; population based; prevent; research study; uptake; virus host interaction; virus pathogenesis

DESCRIPTION (provided by applicant): Over 170 million people world-wide are infected with hepatitis C virus (HCV), a major cause of liver cirrhosis and hepatocellular carcinoma. Despite the global prevalence of HCV, little is known about how this virus interacts with the host and evades the immune system to establish persistent infection. HCV circulates in infected individuals as a heterogeneous population based on buoyant density and this heterogeneity is due in part to virus association with lipoproteins - including very low density (VLDL), low density (LDL) and high density (HDL) lipoproteins. HCV association with lipoproteins initiates during virus particle assembly, and release of progeny HCV is closely linked to the VLDL secretory pathway. Entry into susceptible cells is also facilitated by interaction with lipoproteins, specifically HDL, which promotes rapid kinetics of entry thought to facilitate escape from neutralizing antibodies. Together, these data support the interaction between HCV and lipoproteins and underscore their role in HCV pathogenesis, however, the precise nature of how these lipoproteins interact with HCV is unknown and the full consequences of their interaction not fully explored. Our overall hypothesis is that HCV virion association with, or incorporation of, host proteins and lipids reduces virus susceptibility to neutralization. Previous studies indicate that lipoprotein interaction is mediated by the HCV surface glycoproteins, E1 and E2. In Specific Aim 1, we will define the specific molecular requirements for lipoprotein (specifically HDL)-HCV interaction using both biochemical assays to dissect the HDL complex and a glycoprotein mutagenesis strategy to identify critical viral residues for binding. Independent of viral infection, lipoproteins have been shown to have immunoregulatory activities, and HDL has specifically been shown to modulate complement activation. Thus, in Specific Aim 2 we will investigate the impact of HCV association with HDL on virus susceptibility to complement-mediated virolysis. Our final Specific Aim is to perform complete proteomic and lipidomic analyses on purified HCV. These data will not only further elucidate the lipoprotein-virus interaction, but will also enable us to catalog the cellular components of the virus particle. Such analyses will provide valuable insight into the composition of the HCV particle and its life cycle.

描述（由申请人提供）：全世界有超过 1.7 亿人感染丙型肝炎病毒 (HCV)，这是肝硬化和肝细胞癌的主要原因。尽管丙型肝炎病毒在全球流行，但人们对这种病毒如何与宿主相互作用并逃避免疫系统以建立持续感染的情况知之甚少。 HCV 作为基于浮力密度的异质群体在感染个体中循环，这种异质性部分归因于病毒与脂蛋白的关联，包括极低密度 (VLDL)、低密度 (LDL) 和高密度 (HDL) 脂蛋白。 HCV 与脂蛋白的结合在病毒颗粒组装过程中启动，子代 HCV 的释放与 VLDL 分泌途径密切相关。与脂蛋白（特别是高密度脂蛋白）的相互作用也有助于进入易感细胞，这促进了快速进入动力学，被认为有助于逃避中和抗体。总之，这些数据支持 HCV 和脂蛋白之间的相互作用，并强调它们在 HCV 发病机制中的作用，然而，这些脂蛋白如何与 HCV 相互作用的确切性质尚不清楚，而且它们相互作用的全部后果尚未得到充分探索。我们的总体假设是，HCV 病毒粒子与宿主蛋白和脂质的关联或掺入会降低病毒对中和的敏感性。先前的研究表明，脂蛋白相互作用是由 HCV 表面糖蛋白 E1 和 E2 介导的。在具体目标 1 中，我们将使用生化检测来剖析 HDL 复合物和糖蛋白诱变策略来确定脂蛋白（特别是 HDL）-HCV 相互作用的具体分子要求，以识别关键的病毒残基进行结合。脂蛋白已被证明具有免疫调节活性，与病毒感染无关，而 HDL 已被证明可以调节补体激活。因此，在具体目标 2 中，我们将研究 HCV 与 HDL 的关联对病毒对补体介导的病毒溶解的敏感性的影响。我们的最终具体目标是对纯化的 HCV 进行完整的蛋白质组学和脂质组学分析。这些数据不仅将进一步阐明脂蛋白-病毒的相互作用，而且还将使我们能够对病毒颗粒的细胞成分进行分类。此类分析将为了解 HCV 颗粒的组成及其生命周期提供有价值的见解。