Role of the Viral Capsid in HIV-1 Integration

病毒衣壳在 HIV-1 整合中的作用

• 批准号: 9977928
• 负责人: Chandravanu Dash
• 金额: $ 36.55万
• 依托单位: MEHARRY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9977928
• 关键词: AIDS/HIV problem; Antiviral Agents; Antiviral Therapy; Area; Binding; Biochemical; Biochemistry; Biological Assay; Biology; Capsid; Capsid Proteins; Cell Nucleus; Cell fusion; Cells; Cellular Membrane; Clinic; Clinical; Complex; Coupling; Cultured Cells; Cytoplasm; DNA Integration; Data; Dependence; Development; Disease Management; Drug Targeting; Drug usage; Environment; Exhibits; Face; Fractionation; Goals; HIV; HIV Infections; HIV-1; In Vitro; Infection; Integrase; Integration Host Factors; Knowledge; Life Cycle Stages; Link; Measures; Mediating; Membrane; Modeling; Morphology; Nuclear; Nuclear Pore Complex; Pathway interactions; Play; Proteins; Proviruses; Reaction; Regimen; Reporting; Research; Resistance; Reverse Transcription; Role; Sucrose; Testing; United States National Institutes of Health; Variant; Viral; Virus; Virus Integration; antiretroviral therapy; base; design; drug development; inhibitor/antagonist; multidisciplinary; mutant; next generation; novel; pandemic disease; side effect; skills; small molecule; small molecule inhibitor; therapeutic target; viral DNA; viral resistance

ABSTRACT The ongoing HIV pandemic has resulted in ~1.2 million infections in the US and ~37 million infections worldwide. In spite of considerable progress in HIV/AIDS research, anti-retroviral therapy (ART) remains the only treatment option for HIV-1 infection. While ART has been highly effective in controlling the virus and making HIV infection a manageable disease, the drugs used in the ART regimen are expensive, cause side effects, and face viral resistance. Thus, there is an urgent need for continued development of drugs against novel cellular and viral targets. HIV-1 capsid protein (CA) is an important viral therapeutic target that is currently clinically unexploited. Recently, Gilead developing a highly potent CA-targeting inhibitor as next generation of antivirals. However, successful transition of these CA-specific inhibitors to the clinic require clear understanding of the anti-viral mechanisms and the delineating CA’s role in HIV-1 infection. Our studies are designed to understand the mechanism by which HIV-1 CA controls viral integration. HIV enters the target cell by fusion of the viral membrane with the cellular membrane, releasing the viral capsid core into the cytoplasm of the target cell. Functional studies of HIV-1 variants indicate that the proper assembly, morphology, and stability of the capsid core are all essential for HIV-1 infectivity. While it is well established that HIV-1 CA facilitates reverse transcription, recent data shows that CA is also a key determinant of the ability of HIV-1 to enter the nucleus of the target cell. In particular, CA is genetically and functionally implicated in nuclear entry of the reverse transcribed provirus, by mediating interactions with cellular factors. A key knowledge gap is the role of CA in viral integration-a critical post-nuclear entry step of HIV-1 infection. We hypothesize that HIV-1 CA protein directly influences viral DNA integration. To test this hypothesis we propose three specific aims: Aim 1. To define the effects of capsid stability on HIV-1 preintegration complex (PIC) activity and viral DNA integrity. Aim 2. To determine whether integration activity is dependent on CA levels in the PICs. Aim 3. To determine the role of known capsid-binding host proteins in defining PIC-associated CA levels and PIC activity. To test a direct link between HIV-1 CA and viral DNA integration, we have developed a novel model by coupling a biochemical approach that quantitatively measures PIC-associated integration activity to the use of the CA-specific inhibitor as probes. In addition, we have assembled a multidisciplinary team with expertise in PIC biochemistry (Dash), capsid biology (Aiken) and retroviral integration (Engelman). Therefore, the proposed studies will generate new knowledge on the mechanism by which CA regulates HIV-1 integration and define the antiviral effects of CA-inhibitors to facilitate the development of novel CA-based anti-viral therapies (a High-Priority HIV/AIDS research area of the NIH).

抽象的 正在进行的艾滋病毒大流行导致美国约120万次感染和约3700万感染 全世界。尽管在艾滋病毒/艾滋病研究中考虑了进展，但抗逆转录病毒疗法（ART）仍然是 HIV-1感染的唯一治疗选择。虽然艺术在控制病毒方面非常有效 使艾滋病毒感染成为可控制的疾病，艺术方案中使用的药物很昂贵，原因是 效果，面对病毒抗性。那迫切需要继续开发针对的毒品 新型细胞和病毒靶标。 HIV-1衣壳蛋白（CA）是重要的病毒治疗靶点 目前临床意外。最近，吉利德（Gilead 产生抗病毒药。但是，这些CA特异性抑制剂成功过渡到诊所需要清晰 了解抗病毒机制和描述CA在HIV-1感染中的作用。我们的研究是 旨在了解HIV-1 CA控制病毒整合的机制。 HIV通过融合病毒膜与细胞膜进入靶细胞，释放 病毒式衣壳核进入靶细胞的细胞质。 HIV-1变体的功能研究表明 衣壳核的适当组装，形态和稳定性对于HIV-1感染都是必不可少的。虽然是 良好确定HIV-1 CA的最爱逆转录，最近的数据表明CA也是关键 HIV-1进入靶细胞核的能力的决定因素。特别是，CA是遗传性的， 通过调解与 细胞因子。关键知识差距是CA在病毒整合中的作用 - 关键的核后进入步骤 HIV-1感染。我们假设HIV-1 CA蛋白直接影响病毒DNA整合。测试 我们提出了这三个特定目的：目的1。定义衣壳稳定性对HIV-1的影响 预一体化复合物（PIC）活性和病毒DNA完整性。目标2。确定是否集成活动 取决于图片中的CA水平。目的3。确定已知的胶囊结合宿主蛋白在 定义与PIC相关的CA水平和PIC活性。测试HIV-1 CA和病毒DNA之间的直接联系 整合，我们通过耦合生化方法来开发一种新型模型 衡量与PIC相关的整合活性与CA特异性抑制剂的使用。另外，我们 已经组建了一个具有PIC生物化学专业知识（DASH），CAPSID生物学（Aiken）和 逆转录病毒整合（恩格曼）。因此，拟议的研究将产生有关 CA调节HIV-1整合并定义CA抑制剂的抗病毒作用以促进HIV-1的机制 新型基于CA的抗病毒疗法的发展（NIH的高优先艾滋病毒/艾滋病研究领域）。