Isolation of Genetic Suppressor Elements Against Hepatitis C Virus

抗丙型肝炎病毒基因抑制元件的分离

• 批准号: 7708619
• 负责人: Zhilei Chen
• 金额: $ 22.48万
• 依托单位: TEXAS ENGINEERING EXPERIMENT STATION
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7708619
• 关键词: Address; Antiviral Agents; Be++ element; Beryllium; Binding; Biological Process; Cell Communication; Cell Culture Techniques; Cell Line; Cell Survival; Cells; Cessation of life; Chronic Hepatitis C; Cirrhosis; Classification; Clinical Trials; Complex; Complex Mixtures; Cultured Cells; Development; Drug Delivery Systems; Drug Metabolic Detoxication; Effectiveness; Elements; Escape Mutant; Future; Genes; Genetic; Genome; Genomic Library; Goals; Harvest; Health; Hepatitis C; Hepatitis C Antiviral; Hepatitis C Therapy; Hepatitis C virus; Individual; Interferons; Lead; Left; Lentivirus Vector; Libraries; Life Cycle Stages; Liver; Liver Failure; Mediating; Membrane Proteins; Methodology; Molecular; Nonstructural Protein; Nucleic Acids; Peptide Hydrolases; Peptide Leader Sequences; Peptides; Pharmaceutical Preparations; Polymerase; Preclinical Drug Evaluation; Primary carcinoma of the liver cells; Procedures; Process; Proteins; Public Health; Research; Ribavirin; Risk; Severities; Staging; System; Taxes; Therapeutic Agents; Time; Viral; Virus; Virus Inhibitors; Virus Replication; Work; anti-hepatitis C; base; cost; cytotoxicity; design; drug candidate; drug development; effective therapy; frontier; genetic element; hepatoma cell; inhibitor/antagonist; innovation; interest; liver transplantation; member; mimetics; novel; particle; programs; protective effect; public health relevance; response; small hairpin RNA; viral resistance; virus genetics; virus host interaction

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) infection is a major public health problem, putting infected individuals (~180 million worldwide) at risk of developing cirrhosis, hepatocellular carcinoma, and liver failure. Chronic hepatitis C is the leading cause for liver transplantation. Current standard interferon-based therapy, a costly and time-consuming process, has only a ~50% cure rate, and no anti-HCV drugs have yet been approved for hepatitis C therapy. Despite the promise shown by HCV-specific proteases and polymerases as drug targets, the rapid emergence of viral resistance indicates that additional targets and combinations of antivirals will be necessary for effective treatment. We propose to isolate genetic suppressor elements (GSEs) from a library comprising a fragmented HCV genome which could be used both as potent anti-HCV therapeutic agents and as probes to identify and validate new targets for further drug screening. GSEs are nucleic acid or protein/peptide molecules derived from a gene or genome that act as transdominant inhibitors of a particular biological function through a variety of mechanisms, which includes binding to and blocking essential interaction surfaces for protein activity. In order to identify GSEs from within the HCV genome that exert inhibitory activity against HCV, a novel function-based selection system will be developed and implemented. Briefly, a fragmented HCV genome will be delivered to a hepatoma derivative cell line that is sensitive to a cytopathic effect exerted by HCV. The cells containing the HCV-derived genetic fragments will be subjected to a cytopathic challenge by exogenously administered cell culture-derived HCV (HCVcc) infectious particles, and cells surviving this HCV challenge will be enriched in GSEs that exert an inhibitory effect against HCV. Iterative application of this selection procedure will result in the identification of highly potent GSEs that protect cells against HCV infection and cytotoxicity. Preliminary studies will evaluate the degree of the protective effect conferred by the identified GSEs against HCV-mediated cytotoxicity, and the stage in the HCV life cycle at which the anti-HCV effect of the GSEs is exerted. Future studies originating from the identified GSEs are expected to reveal the molecular basis of the anti-HCV GSE activity, thus providing new leads for anti-HCV drug development. It is expected that the GSEs identified from the proposed research, and molecular mimetics derived therefrom, will inhibit HCV infection/propagation through diverse mechanisms, including the inhibition of virus-host cell interactions, thus contributing to the urgent quest for new and effective HCV antivirals on multiple fronts. PUBLIC HEALTH RELEVANCE: Infection by hepatitis C virus is a serious global health problem that causes numerous debilitating liver conditions. The current treatment regime for hepatitis C is time-consuming, expensive, and often ineffective, creating an urgent need for new and effective drugs. Novel anti-hepatitis C genetic suppressor elements isolated from this research will serve both as hepatitis C drugs, and as keys to open doors to new avenues of research in hepatitis C antiviral development.

描述(申请人提供)：丙型肝炎病毒(丙型肝炎病毒)感染是一个主要的公共卫生问题，使感染者(全球约1.8亿人)面临发展为肝硬变、肝细胞癌和肝功能衰竭的风险。慢性丙型肝炎是肝移植的主要原因。目前基于干扰素的标准治疗是一个昂贵和耗时的过程，治愈率只有50%左右，而且还没有抗丙型肝炎药物被批准用于丙型肝炎的治疗。尽管丙型肝炎病毒特异的蛋白酶和聚合酶有望成为药物靶点，但病毒耐药性的迅速出现表明，有效的治疗需要更多的靶点和抗病毒药物的组合。我们建议从包含一个片段的丙型肝炎病毒基因组的文库中分离出遗传抑制元件(GSE)，该文库既可以作为有效的抗丙型肝炎病毒治疗药物，也可以作为探针来识别和验证进一步药物筛选的新靶点。GSE是来源于基因或基因组的核酸或蛋白质/多肽分子，通过各种机制作为特定生物功能的跨显性抑制物，包括结合和阻断蛋白质活性的基本相互作用面。为了从丙型肝炎病毒基因组中识别对丙型肝炎病毒具有抑制活性的GSE，将开发和实现一种新的基于功能的选择系统。简而言之，一个片段的丙型肝炎病毒基因组将被传递给一个对丙型肝炎病毒所产生的细胞病变效应敏感的肝癌衍生细胞系。含有丙型肝炎病毒基因片段的细胞将受到外源性细胞培养衍生丙型肝炎病毒(HCVcc)感染颗粒的细胞病变攻击，而在这种丙型肝炎病毒挑战中存活的细胞将富含对丙型肝炎病毒起抑制作用的GSE。反复应用这一选择程序将导致高度有效的GSE的鉴定，以保护细胞免受丙型肝炎病毒感染和细胞毒性。初步研究将评估已鉴定的GSE对丙型肝炎病毒介导的细胞毒性的保护作用程度，以及在丙型肝炎病毒生命周期中发挥抗丙型肝炎病毒作用的阶段。来自已鉴定的GSE的未来研究有望揭示抗丙型肝炎病毒GSE活性的分子基础，从而为抗丙型肝炎药物的开发提供新的线索。预计从拟议的研究中确定的GSE及其衍生的分子模拟物将通过不同的机制(包括抑制病毒与宿主细胞的相互作用)抑制丙型肝炎病毒的感染/繁殖，从而在多个方面促进对新的有效的丙型肝炎病毒抗病毒药物的迫切探索。公共卫生相关性：丙型肝炎病毒感染是一个严重的全球健康问题，会导致许多肝脏虚弱。目前丙型肝炎的治疗方案耗时、昂贵，而且往往无效，因此迫切需要新的有效药物。从这项研究中分离出的新型抗丙型肝炎基因抑制因子将既用作丙型肝炎药物，也将成为打开丙型肝炎抗病毒开发研究新途径的钥匙。