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（在全球范围内约有1.8亿）面临患肝硬化，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。当前的标准干扰素治疗，这是一个昂贵且耗时的过程，仅治愈率约50％，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标的有希望，但病毒抗性的迅速出现表明，有效治疗需要其他靶标和组合。我们建议将遗传抑制元件（GSE）与包含碎片的HCV基因组组成的文库分离，该图书馆既可以用作有效的抗HCV治疗剂，又可以用作识别和验证对进一步药物筛查的新目标的探针。 GSE是源自源自基因或基因组的核酸或蛋白质/肽分子，通过多种机制起到特定生物学功能的跨性抑制剂，其中包括与蛋白质活性的结合和阻断必需的相互作用表面。为了从HCV基因组内识别GSE，对HCV发挥抑制活性，将开发和实施一个新型的基于功能的选择系统。简而言之，将碎片的HCV基因组传递到肝癌衍生细胞系，该细胞系对HCV施加的细胞病变效应敏感。含有HCV衍生的遗传片段的细胞将受到外源给予细胞培养的衍生的HCV（HCVCC）感染性颗粒的细胞病变挑战，并且在这种HCV挑战中幸存的细胞将在GSE中富含对HCV的抑制作用。该选择程序的迭代应用将导致鉴定高度有效的GSE，以保护细胞免受HCV感染和细胞毒性的影响。初步研究将评估已鉴定的GSE赋予HCV介导的细胞毒性的保护作用程度，以及在HCV寿命中施加了GSE抗HCV效应的阶段。源自确定的GSE的未来研究预计将揭示抗HCV GSE活性的分子基础，从而为抗HCV药物开发提供新的铅。可以预期，从拟议的研究中鉴定出的GSE以及从中得出的分子模仿物将通过各种机制抑制HCV感染/传播，包括抑制病毒宿主细胞相互作用，从而有助于迫切寻求新的和有效的HCV抗病毒药物。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，会导致许多使肝脏疾病衰弱。当前针对丙型肝炎的治疗方案是耗时，昂贵且通常无效的，这迫切需要新的有效药物。从这项研究中分离出的新型抗肝炎遗传抑制元件将既可以用作丙型肝炎药物，又是为丙型肝炎抗病毒发育研究的新途径打开大门的关键。