VIF Anatagonists: Antiviral Evaluation in Vitro, Inhibitor resistance and Mole

VIF 拮抗剂：体外抗病毒评价、抑制剂耐药性和葡萄胎

• 批准号: 7597047
• 负责人: Mario Stevenson
• 金额: $ 21.5万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7597047
• 关键词: Antiviral Agents; Attenuated; Biochemistry; Biological Assay; Cell Line; Cell Survival; Cells; Class; Complementary DNA; Cytidine Deaminase; Defective Viruses; Encephalitis; Evaluation; Exhibits; Experimental Models; Exposure to; Fluorescence; Genetic Polymorphism; Goals; Green Fluorescent Proteins; HIV; HIV-1; In Vitro; Individual; Infection; Laboratories; Lead; Lymphoid; Macaca; Mediating; Melanocytic nevus; Modeling; Mole the mammal; Molecular; Monkeys; Mutation; Numbers; Pathogenesis; Pathogenicity; Personal Satisfaction; Pharmacology; Phenotype; Physiological; Plasma; Primate Lentiviruses; Proteins; RNA; Rana; Reporting; Resistance; SIV; Screening procedure; Side; Specificity; Viral; Virion; Virus; analog; base; cellular targeting; design; in vivo; inhibitor/antagonist; insight; macrophage; mutant; vif Genes; viral RNA

Vif is essential to the replication of primate lentiviruses such as HIV-1 and SIV in vivo. In the last two years, a considerable amount of insight has been gained into the mechanism by which Vif supports viral replication. Vif counteracts the antiviral effects of the cytidine deaminases apobec 3G and 3F. To inhibit viral replication, these cytidine deaminases must be packaged within virions where they edit the minus strand of nascent viral cDNA in the target cell. Vif counteracts the antiviral activity of apobec 3G and 3F by promoting their proteasomal degradation to the extent that there are insufficient levels for packaging into virions. The overall goal of this project is to identify and develop Vif antagonists that can be used to assess the antiviral impact of Vif antagonism in vivo. Project 2 will prioritize Vif antagonists that emerge from the inhibitor screen of Project 1 on the basis of antiviral activity and specificity. Since the most favorable Vif antagonists will be evaluated in the macaque model of SIV pathogenesis and SIV-induced encephalitis, antiviral activity will be evaluated in cells that are relevant to CNS infection namely, primary macrophages. Project 2 will also assess mechanisms of inhibitor escape in vitro and in vivo. The major activities of Project 2 include: Evaluation of antiviral activity and specificity in cells that express apobec (non-permissive cells) and cells that do not express apobec (permissive cells). Evaluation of antiviral activity in CNS-relevant target cells in vitro, namely, primary macrophages. Identification of the mechanism of Vif antagonism by assessing impact of Vif antagonists on virion encapsidation of apobec and on apobec-mediated editing of viral cDNA. Identification of mutations in Vif that confer inhibitor resistance in vitro and in vivo. Collectively, these studies will allow us to identify the most promising lead Vif antagonists and prioritize the selection of inhibitors that will be evaluated in an experimental model of SIV replication and SIV-induced encephalitis in macaques.

Vif 对于灵长类慢病毒（例如 HIV-1 和 SIV）的体内复制至关重要。在过去的两年里， 人们对 Vif 支持病毒复制的机制有了相当多的了解。 Vif 抵消胞苷脱氨酶 apobec 3G 和 3F 的抗病毒作用。为了抑制病毒复制， 这些胞苷脱氨酶必须包装在病毒颗粒内，在那里它们编辑新生病毒的负链 靶细胞中的 cDNA。 Vif 通过促进 apobec 3G 和 3F 的抗病毒活性来抵消它们 蛋白酶体降解程度不足以包装成病毒体。 该项目的总体目标是识别和开发可用于评估 Vif 拮抗剂 Vif 拮抗体内抗病毒作用。项目 2 将优先考虑从 根据抗病毒活性和特异性对项目1进行抑制剂筛选。自从最优惠的Vif 将在 SIV 发病机制和 SIV 诱发的脑炎的猕猴模型中评估拮抗剂， 将在与中枢神经系统感染相关的细胞（即初级巨噬细胞）中评估抗病毒活性。 项目 2 还将评估体外和体内抑制剂逃逸机制。项目主要活动 2 包括： 评估表达 apobec 的细胞（非许可细胞）的抗病毒活性和特异性 不表达 apobec 的细胞（许可细胞）。 体外评估中枢神经系统相关靶细胞（即初级巨噬细胞）的抗病毒活性。 通过评估 Vif 拮抗剂对病毒粒子的影响来鉴定 Vif 拮抗机制 apobec 的衣壳化和 apobec 介导的病毒 cDNA 编辑。 鉴定在体外和体内赋予抑制剂抗性的 Vif 突变。 总的来说，这些研究将使我们能够确定最有希望的先导 Vif 拮抗剂，并优先考虑 选择将在 SIV 复制和 SIV 诱导的实验模型中进行评估的抑制剂 猕猴脑炎。