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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：丙型肝炎病毒 (HCV) 感染是一个主要的公共卫生问题，使感染者（全球约 1.8 亿）面临发展为肝硬化、肝细胞癌和肝衰竭的风险。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过程昂贵且耗时，治愈率仅为 50% 左右，并且尚未批准用于丙型肝炎治疗的抗 HCV 药物。尽管HCV特异性蛋白酶和聚合酶作为药物靶点显示出良好的前景，但病毒耐药性的迅速出现表明，为了有效治疗，需要额外的靶点和抗病毒药物组合。我们建议从包含HCV基因组碎片的文库中分离出遗传抑制元件（GSE），该元件既可以用作有效的抗HCV治疗剂，也可以用作探针来识别和验证进一步药物筛选的新靶点。 GSE 是源自基因或基因组的核酸或蛋白质/肽分子，通过多种机制充当特定生物功能的反显性抑制剂，其中包括结合和阻断蛋白质活性的必需相互作用表面。为了从 HCV 基因组中识别对 HCV 具有抑制活性的 GSE，将开发和实施一种新的基于功能的选择系统。简而言之，片段化的HCV基因组将被递送至对HCV所施加的细胞病变效应敏感的肝癌衍生细胞系。含有HCV衍生的遗传片段的细胞将受到外源施用的细胞培养衍生的HCV (HCVcc)感染性颗粒的细胞病变攻击，并且在该HCV攻击中幸存的细胞将富含对HCV发挥抑制作用的GSE。该选择程序的迭代应用将鉴定出高效的 GSE，可保护细胞免受 HCV 感染和细胞毒性。初步研究将评估已鉴定的GSE对HCV介导的细胞毒性的保护作用的程度，以及GSE在HCV生命周期中发挥抗HCV作用的阶段。未来基于已鉴定的 GSE 的研究有望揭示抗 HCV GSE 活性的分子基础，从而为抗 HCV 药物开发提供新的线索。预计从拟议研究中鉴定出的 GSE 以及由此衍生的分子模拟物将通过多种机制抑制 HCV 感染/传播，包括抑制病毒与宿主细胞的相互作用，从而有助于迫切寻求新型有效的 HCV 抗病毒药物在多个方面。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，会导致多种肝脏疾病。目前的丙型肝炎治疗方案耗时、昂贵且常常无效，因此迫切需要新的有效药物。从这项研究中分离出来的新型抗丙型肝炎基因抑制元件将既可用作丙型肝炎药物，又可作为打开丙型肝炎抗病毒开发研究新途径之门的钥匙。