Identifying determinants of Hepatitis C Virus (HCV) tropism

确定丙型肝炎病毒 (HCV) 趋向性的决定因素

• 批准号: 7201634
• 负责人: TATJANA DRAGIC
• 金额: $ 35.63万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7201634
• 关键词: Alleles; Biochemical; Biological Assay; Biology; CD209 gene; CD81 gene; Cell Communication; Cell Line; Cell Surface Receptors; Cell surface; Cells; Cellular Tropism; Chronic Hepatitis; Cirrhosis; Clinical; Cloning; Complementary DNA; Complex; Development; Disease Progression; Endoplasmic Reticulum; Exhibits; Experimental Models; Foundations; Gene Transfer; Genetic Polymorphism; Glycoproteins; Goals; Hepatitis C; Hepatitis C virus; Hepatocyte; Human; Human Cell Line; In Vitro; Individual; Infection; L Forms; Licensing; Liver; Mediating; Membrane Fusion; Molecular; Natural History; Organ; Pathogenesis; Peripheral Blood Mononuclear Cell; Play; Predisposition; Prevention therapy; Primary carcinoma of the liver cells; Production; Property; Proteins; Public Health; Range; Reporter; Research; Retroviridae; Risk; Role; Serum; Structure; System; Techniques; Testing; Toxic effect; Tropism; Vaccines; Viral; Virion; Virus; Virus Diseases; Virus Replication; Work; cell type; in vivo; novel; receptor; response; tissue tropism; virus culture; virus envelope; virus tropism

DESCRIPTION (provided by applicant): It is estimated that 170 million people worldwide are infected with the Hepatitis C virus (HCV) and are at risk of developing chronic hepatitis or cirrhosis, the latter often leading to hepatocellular carcinoma. There is currently no vaccine and licensed therapies are associated with modest efficacies and significant toxicities. Despite the urgency of this worldwide public health problem, our basic understanding of HCV replication and pathogenesis remains poor due to a lack of key experimental models. For example, difficulties in culturing the virus in vitro and expressing native, fusogenic envelope glycoproteins have greatly limited studies of HCV tropism and entry. These are critical aspects of viral biology because the host range and pathogenesis of enveloped virus infection is largely determined by the selective interaction of viral envelope glycoproteins with cell-surface receptors. A major goal in HCV research is to understand how HCV targets the liver and by what mechanism it enters host cells. Recently, a major breakthrough in the field has been the development of retroviruses pseudotyped with HCV envelope glycoproteins that specifically mediate infection of primary hepatocytes, as well as certain other human cells. We will use this new experimental system to study HCV entry into target cells. Alterations in naturally occurring HCV envelope glycoproteins may predicate differences in receptor usage and target cell tropism in vivo. To investigate the range of HCV cellular tropism, pseudotypes incorporating envelope glycoproteins from clinical HCV isolates will be tested for their ability to enter relevant primary cells and cell lines. A functional cDNA cloning approach will be used to identify cell-surface receptors that specifically mediate HCV entry into different target cells. However, these receptors may be ubiquitously expressed and HCV targeting to different cell types may be determined by another mechanism. We recently demonstrated that L-SIGN and DC-SIGN are specific HCV-capture receptors and we will explore whether they mediate infection of target cells in trans, thereby determining HCV tropism. The major objective of our work is to identify the basic protein interactions that mediate HCV tropism, which will serve as a foundation for detailed structure/function analyses of HCV receptors and envelope glycoproteins.

描述(申请人提供)：据估计，全球有1.7亿人感染丙型肝炎病毒(丙型肝炎病毒)，并面临发展为慢性肝炎或肝硬变的风险，后者往往导致肝细胞癌。目前还没有疫苗，获得许可的治疗方法与适度的疗效和严重的毒性有关。尽管这一全球公共卫生问题迫在眉睫，但由于缺乏关键的实验模型，我们对丙型肝炎病毒复制和发病机制的基本了解仍然很差。例如，在体外培养病毒和表达天然融合的包膜糖蛋白的困难极大地限制了对丙型肝炎病毒趋向性和进入的研究。这些都是病毒生物学的关键方面，因为包膜病毒感染的宿主范围和致病机制在很大程度上取决于病毒包膜糖蛋白与细胞表面受体的选择性相互作用。丙型肝炎病毒研究的一个主要目标是了解丙型肝炎病毒是如何攻击肝脏的，以及它是通过什么机制进入宿主细胞的。最近，该领域的一项重大突破是发展了以丙型肝炎病毒包膜糖蛋白为假型的逆转录病毒，这种逆转录病毒特异性地介导原代肝细胞以及某些其他人类细胞的感染。我们将使用这个新的实验系统来研究丙型肝炎病毒进入靶细胞的情况。自然产生的丙型肝炎病毒包膜糖蛋白的变化可能预示着体内受体使用和靶细胞趋向性的差异。为了研究丙型肝炎病毒细胞嗜性的范围，将测试从临床分离的丙型肝炎病毒分离株中含有包膜糖蛋白的伪型，以了解它们进入相关原代细胞和细胞系的能力。一种功能性的cDNA克隆方法将被用来识别细胞表面受体，这些受体特异性地介导丙型肝炎病毒进入不同的靶细胞。然而，这些受体可能普遍表达，丙型肝炎病毒针对不同细胞类型的靶向可能由另一种机制决定。我们最近证实了L-SIGN和DC-SIGN是特异性的丙型肝炎病毒捕获受体，我们将探讨它们是否介导反式靶细胞的感染，从而确定丙型肝炎病毒的趋向性。我们工作的主要目标是确定介导丙型肝炎病毒趋向性的基本蛋白质相互作用，这将作为详细分析丙型肝炎病毒受体和包膜糖蛋白结构/功能的基础。