Drug development for Vif-APOBEC3G in HIV-1/AIDS

Vif-APOBEC3G 治疗 HIV-1/AIDS 的药物开发

• 批准号: 7152570
• 负责人: David Kabat
• 金额: $ 36.01万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7152570
• 关键词: Acquired Immunodeficiency Syndrome; Affinity; Anti-Retroviral Agents; Antiviral Agents; Binding; Biological Assay; Cell Line; Cells; Cullin 5 Protein; Cytidine Deaminase; Dependency; Development; Enzyme-Linked Immunosorbent Assay; Evaluation; Funding; Goals; HIV-1; Lead; Lymphocyte; Mammalian Cell; Measures; Methods; NIH Program Announcements; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Pichia; Play; Polyubiquitination; Preclinical Drug Evaluation; Process; Proteins; Protocols documentation; Request for Applications; Research Personnel; Resources; Role; Screening procedure; Series; Standards of Weights and Measures; System; T-Lymphocyte; Therapeutic; Transcript; Translating; Viral; Yeasts; acrosome stabilizing factor; assay development; base; cytotoxic; design; drug development; high throughput screening; inhibitor/antagonist; insight; macrophage; novel; paralogous gene; programs; response; small molecule; small molecule libraries; vif Gene Products

DESCRIPTION (provided by applicant): Recent studies by ourselves and others have substantially elucidated the role of the HIV-1 encoded viral infectivity factor (Vif) in neutralizing a potent antiviral system that occurs in lymphocytes and macrophages and some leukemic T cell lines. This antiviral system principally involves the cytidine deaminases APOBEC3G (A3G) and/or ASF, which are incorporated into HIV-1 cores where they lethally hypermutate newly synthesized viral reverse transcripts. Vif binds to A3G and ASF and induces their polyubiquitination and degradation, thereby eliminating them from infected cells and precluding their incorporation into HIV-1 progeny. Although researchers have used one Vif as a standard, HIV-1 Vifs are highly divergent and we have found that natural HIV-1 isolates encode Vifs that bind promiscuously to all APOBEC3 paralogs but have widely distinctive effects on their concentrations. In addition, the APOBECSs are coexpressed in different amounts and proportions in HIV-1 susceptible cells, and they broadly heterooligomerize to form a collaborative and inducible antiviral network. Our results suggest that Vif diversity is partly an adaptive response to the diversity of the APOBEC3 network in different cells and compartments, that it may play a critical role in HIV-1 pathogenesis, and that it should also be considered in anti-Vif drug development. Based on these results and on substantial preliminary evidence, we propose three substantial and synergistic aims: (1) Develop a robust series of screening platforms for identification of small molecules that inhibit diverse Vifs within mammalian cells. (2) We found that HIV-1 Vif is made in yeast as a soluble protein that associates with A3G. Produce substantial amounts of Vif, and its A3G-binding subdomain and A3G in the yeast Pichia pastoris as a resource to analyze inhibitor mechanisms, and to support the high throughput screening system of aim 3. (3) Optimize ELISA assays to screen and analyze compounds that inhibit Vif- A3G binding and to measure affinities of diverse Vifs for A3G and other cytidine deaminases. This program builds on recent insights to develop and to optimize novel inhibitor screening approaches and investigational resources for AIDS, as an essential prelude to high throughput screening for effective anti-Vif therapeutics.

描述（由申请人提供）：我们自己和其他人的最新研究已大大阐明了HIV-1编码的病毒感染因子（VIF）在中和淋巴细胞和巨噬细胞和一些白血病T细胞系中发生有效的抗病毒系统中的作用。该抗病毒系统主要涉及胞苷脱氨酶APOBEC3G（A3G）和/或ASF，它们掺入HIV-1核心中，在这些核心中，它们致命地过度挤压了新合成的病毒逆转录。 VIF与A3G和ASF结合，并诱导它们的多泛素化和降解，从而从感染的细胞中消除它们，并排除其掺入HIV-1后代。尽管研究人员已使用一种VIF作为标准，但HIV-1 VIF具有很高的分歧，我们发现天然HIV-1分离株编码与所有APOBEC3旁系同源物结合但对其浓度具有广泛独特的影响的VIF。此外，Apobecs在HIV-1易感细胞中以不同的量和比例共表达，并且它们广泛地异性构成形成协作和诱导的抗病毒网络。我们的结果表明，VIF多样性部分是对不同细胞和隔室中APOBEC3网络多样性的一种适应性反应，它可能在HIV-1发病机理中起关键作用，并且在抗VIF药物开发中也应考虑。基于这些结果和实质性的初步证据，我们提出了三个实质性和协同的目的：（1）开发一系列可靠的筛选平台，用于鉴定抑制哺乳动物细胞中不同VIF的小分子。 （2）我们发现HIV-1 VIF是在酵母中作为一种与A3G相关的可溶性蛋白。 Produce substantial amounts of Vif, and its A3G-binding subdomain and A3G in the yeast Pichia pastoris as a resource to analyze inhibitor mechanisms, and to support the high throughput screening system of aim 3. (3) Optimize ELISA assays to screen and analyze compounds that inhibit Vif- A3G binding and to measure affinities of diverse Vifs for A3G and other cytidine deaminases.该计划以最新的见解为基础，以开发和优化新型抑制剂筛查方法和艾滋病的研究资源，这是对有效抗VIF治疗剂的高吞吐量筛查的重要序曲。