High Throughput in vitro Hepatitis C Virus Screening

高通量体外丙型肝炎病毒筛查

• 批准号: 7276043
• 负责人: Susan L. Uprichard
• 金额: $ 27.24万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7276043
• 关键词: Adverse effects; Animal Model; Antiviral Agents; Area; Binding; Biological Assay; Biology; Cells; Development; HCV screening; Hepatitis; Hepatitis C; Hepatitis C virus; In Vitro; Infection; Infection prevention; Investigation; Life Cycle Stages; Light; Liver Cirrhosis; Liver diseases; Patients; Primary carcinoma of the liver cells; Public Health; Replicon; Research; Screening procedure; Specificity; Staging; System; Toxic effect; United States National Institutes of Health; Vaccines; Viral; Virus; Virus Diseases; base; drug discovery; hepatitis C virus NS3 protein; high throughput screening; novel; tissue culture; tool

DESCRIPTION (provided by applicant): With more than 170 million people infected worldwide, Hepatitis C Virus (HCV) has emerged as a significant public health burden. Unfortunately, there is no vaccine available to prevent this infection, and the only approved treatment has significant toxic side effects and is only effective in a subset of patients. Notably research efforts to understand HCV infection have been hindered by the lack of robust tissue culture infection systems and small animal models. Recently however, we and others have developed a robust in vitro HCV infection system, which should finally enable investigation of all stages of viral infection. In light of the immediate need to elucidate the biology of this virus and identify alternative HCV treatment option, the objective of this application is to develop our in vitro HCV infection system for high throughput screening (HTS) of potential antiviral compounds. The cell-based infection assay proposed would provide several advantages over the currently available HTS approaches being used for HCV drug discovery because it recapitulates the entire viral life cycle and does not restrict screening to any specific predetermined viral target(s). Thus, this HTS infection assay would have the potential to not only provide novel leads for drug discovery efforts, but also to identify new compounds with unique mechanisms of action that can be used as investigational tools to study HCV infection. Accordingly, this study has 3 Specific Aims: 1) Implement, optimize, and validate a FRET-based NS3 protease HCV infection assay amenable to HTS. 2) Implement, optimize, and validate secondary screens that can be used to prioritize initial hits based on specificity, efficacy, and toxicity. 3) Verify the high throughput capacity of the system by performing preliminary, proof-of-principle screens and analyzing hits to target and/or mechanism of action.

描述(申请人提供)：全球有超过1.7亿人感染丙型肝炎病毒(丙型肝炎病毒)，已成为重大的公共卫生负担。不幸的是，目前还没有预防这种感染的疫苗，唯一被批准的治疗方法具有显著的毒副作用，而且只对一小部分患者有效。值得注意的是，由于缺乏强大的组织培养感染系统和小型动物模型，了解丙型肝炎病毒感染的研究工作一直受到阻碍。然而，最近，我们和其他人开发了一个强大的体外丙型肝炎病毒感染系统，该系统最终应该能够研究病毒感染的所有阶段。 鉴于迫切需要阐明这种病毒的生物学特性并确定替代的丙型肝炎病毒治疗方案，这项应用的目标是开发我们的体外丙型肝炎病毒感染系统，用于高通量筛选潜在的抗病毒化合物(HTS)。与目前用于丙型肝炎病毒药物发现的HTS方法相比，建议的基于细胞的感染分析方法将提供几个优点，因为它概括了整个病毒生命周期，并且不限制对任何特定的预定病毒靶点进行筛选(S)。因此，这种HTS感染分析不仅有可能为药物发现工作提供新的线索，而且还有可能识别具有独特作用机制的新化合物，这些化合物可以用作研究丙型肝炎病毒感染的研究工具。 因此，这项研究有三个具体目标： 1)实现、优化和验证一种基于FRET的、符合HTS的NS3蛋白酶-丙型肝炎病毒感染检测方法。 2)实施、优化和验证可用于根据特异性、有效性和毒性对初始命中进行优先排序的二级筛查。 3)通过执行初步的原则性筛选并分析对目标的命中和/或作用机制，验证系统的高吞吐能力。