Tight Junction: Gate to HCV Tropism, Therapeutics and Pathogenesis

紧密连接：HCV 趋向性、治疗和发病机制的大门

• 批准号: 8101858
• 负责人: TONY WANG
• 金额: $ 31.33万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 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 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8101858
• 关键词: Accounting; Acute Hepatitis; Adverse effects; Affect; Animal Model; Antibodies; Antiviral Agents; Antiviral Therapy; CD81 gene; Cell Culture Techniques; Cell Line; Cell physiology; Cells; Chronic Hepatitis; Cirrhosis; Cleaved cell; Combined Modality Therapy; Complementary DNA; DNA Library; Data; Development; Down-Regulation; Endoplasmic Reticulum; Engineering; Evaluation; Fibroblasts; Future; HIV; Hepatic; Hepatitis C; Hepatitis C virus; Hepatocyte; Human; Immune; Individual; Infection; Infection prevention; Integration Host Factors; Interferons; Laboratories; Lead; Life Cycle Stages; Light; Liver diseases; Maps; Mediating; Molecular; Molecular Biology; Monoclonal Antibodies; Morphology; Mus; NIH 3T3 Cells; Pathogenesis; Pathology; Patients; Peptides; Primary carcinoma of the liver cells; Proteins; Public Health; Recombinants; Reporting; Research; Resistance; Resistance to infection; Ribavirin; Rodent; Screening procedure; Small Interfering RNA; System; Testing; Therapeutic; Tight Junctions; Treatment Protocols; Vaccines; Viral; Virus; Work; base; cellular engineering; claudin-1 protein; env Gene Products; extracellular; global health; hepatitis C virus envelope 2 protein; hepatoma cell; immunoenhancing factor; inhibitor/antagonist; insight; interest; liver biopsy; novel; novel therapeutics; occludin; polyclonal antibody; prevent; public health relevance; receptor; vaccine candidate; virus envelope; virus tropism

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) exacts a heavy toll on global health by causing acute and chronic hepatitis, cirrhosis, as well as hepatocellular carcinoma. Paradoxical to this cold reality, there is no vaccine available to prevent the infection, and combination therapy with pegylated IFN-a and ribavirin is only effective in 40-80% of patients and has severe side effects. Fortunately, with the recently developed infectious clones, we are finally able to dissect the entire viral life cycle at the molecular level. It is conceivable that novel antiviral therapies and treatment regimens relieving HCV associated pathologies will become available within foreseeable future given the accelerated pace of research. As far as what this application concerns, accumulating evidence has highlighted the importance of tight junction (TJ) in HCV entry. Specifically, we and others reported that the tight junction protein occludin (OCLN) is indispensible for HCV envelope protein-dependent entry. Evidence also indicated that OCLN partially accounts for the narrow host range of HCV. However, murine cells expressing all known human (co-)receptors remained resistant to cell culture grown HCV (HCVcc) infection, suggesting either additional human factors are required or dominant inhibitors exist in murine cells. Interestingly, we observed that both OCLN and claudin-1 were downregulated in HCVcc-infected hepatoma cells. Subsequent study suggested that OCLN is increasingly cleaved in HCV infected cells. In light of these findings, we believe novel therapeutics, novel infection systems, and novel insights in understanding HCV-associated pathogenesis may be developed. We hypothesize that antibody against OCLN extracellular loops or OCLN-derived peptide may block the spreading of virus (to be tested in Aim 1); additional host factors are required to support optimal HCV infection in murine cells or restriction factors exist in these cells (to be tested in Aim 2); downregulation of TJ proteins during HCV entry may alter cell physiology (to be tested in Aim 3). We plan to test the hypotheses by (i) assessing the effect of antibodies against OCLN or OCLN-derived peptides on HCV entry. (ii) identifying host factors promoting or restricting HCV infection of murine cells. (iii) analyzing the impact of HCV infection on TJ integrity and hepatocyte functions. PUBLIC HEALTH RELEVANCE: HCV infection poses a serious threat to US public health and there is currently no vaccine available. Completion of the proposed research will shed lights to the development of novel therapeutics, small animal models, and to our understanding of HCV-associated pathogenesis.

描述（由申请人提供）：丙型肝炎病毒（HCV）通过引起急性和慢性肝炎、肝硬化以及肝细胞癌对全球健康造成严重损害。与这一冷酷的现实相反，没有疫苗可用于预防感染，并且聚乙二醇化IFN-α和利巴韦林的联合治疗仅在40-80%的患者中有效，并且具有严重的副作用。幸运的是，随着最近开发的感染性克隆，我们终于能够在分子水平上剖析整个病毒生命周期。可以想象，随着研究步伐的加快，在可预见的未来，新的抗病毒疗法和治疗方案将缓解HCV相关的病理。就本申请所关注的而言，越来越多的证据强调了紧密连接（TJ）在HCV进入中的重要性。具体来说，我们和其他人报道了紧密连接蛋白闭合蛋白（OCLN）是HCV包膜蛋白依赖性进入所必需的。证据还表明，OCLN部分解释了HCV的狭窄宿主范围。然而，表达所有已知人（共）受体的鼠细胞对细胞培养生长的HCV（HCVRNA）感染保持抗性，表明需要额外的人为因素或鼠细胞中存在显性抑制剂。有趣的是，我们观察到OCLN和claudin-1在HCV感染的肝癌细胞中下调。随后的研究表明，OCLN在HCV感染的细胞中越来越多地被切割。根据这些发现，我们相信新的治疗方法，新的感染系统，以及新的见解，在理解HCV相关的发病机制可能会发展。我们假设抗OCLN细胞外环或OCLN衍生肽的抗体可能阻断病毒的传播（在目标1中进行测试）;需要额外的宿主因子来支持鼠细胞中的最佳HCV感染或这些细胞中存在限制因子（在目标2中进行测试）; HCV进入期间TJ蛋白的下调可能改变细胞生理学（在目标3中进行测试）。我们计划通过（i）评估针对OCLN或OCLN衍生肽的抗体对HCV进入的影响来检验假设。(ii)鉴定促进或限制HCV感染鼠细胞的宿主因子。(iii)分析HCV感染对TJ完整性和肝细胞功能的影响。 公共卫生相关性：HCV感染对美国公共卫生构成严重威胁，目前没有可用的疫苗。这项研究的完成将有助于开发新的治疗方法，小动物模型，以及我们对HCV相关发病机制的理解。