Role of the Viral Capsid in HIV-1 Integration

病毒衣壳在 HIV-1 整合中的作用

基本信息

批准号：
10199944
负责人：
Chandravanu Dash
金额：
$ 37万
依托单位：
MEHARRY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-15 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10199944
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

项目摘要

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).

抽象的持续的艾滋病毒大流行已导致美国约 120 万人感染，其中约 3700 万人感染尽管艾滋病毒/艾滋病研究取得了相当大的进展，但抗逆转录病毒疗法（ART）仍然是世界范围内的主要治疗方法。 HIV-1 感染的唯一治疗选择，而 ART 在控制病毒方面非常有效。使艾滋病毒感染成为一种可控制的疾病，昂贵的抗逆转录病毒治疗方案中使用的药物是，原因因此，迫切需要继续开发针对病毒的药物。 HIV-1 衣壳蛋白 (CA) 是一种重要的病毒治疗靶点。目前临床上尚未开发，吉利德公司正在开发一种高效的 CA 靶向抑制剂。然而，这些 CA 特异性抑制剂成功过渡到临床需要明确的条件。了解抗病毒机制并描述 CA 在 HIV-1 感染中的作用。旨在了解 HIV-1 CA 控制病毒整合的机制。 HIV通过病毒膜与细胞膜融合进入靶细胞，释放 HIV-1 变体的功能研究表明，病毒衣壳核心进入靶细胞的细胞质。衣壳核心的正确组装、形态和稳定性对于 HIV-1 的感染性都是至关重要的。众所周知，HIV-1 CA 促进逆转录，最近的数据表明 CA 也是一个关键 HIV-1 进入靶细胞细胞核的能力的决定因素尤其是 CA 是遗传因素和影响因素。通过介导与逆转录原病毒的相互作用，在功能上涉及逆转录原病毒的核进入一个关键的知识缺口是 CA 在病毒整合中的作用，这是病毒进入核后的一个关键步骤。 HIV-1 感染。我们对抗 HIV-1 CA 蛋白直接影响病毒 DNA 整合进行测试。针对这一假设，我们提出了三个具体目标：目标 1. 确定衣壳稳定性对 HIV-1 的影响预整合复合物 (PIC) 活性和病毒 DNA 完整性目的 2. 确定是否具有整合活性。取决于 PIC 中的 CA 水平。目标 3. 确定已知衣壳结合宿主蛋白的作用。定义 PIC 相关的 CA 水平和 PIC 活性以测试 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开发了一种新颖的模型，该方法定量此外，我们还使用 CA 特异性抑制剂作为探针来测量 PIC 相关的整合活性。组建了一支多学科团队，拥有 PIC 生物化学 (Dash)、衣壳生物学 (Aiken) 和因此，拟议的研究将产生关于逆转录病毒整合的新知识。 CA 调节 HIV-1 整合的机制并定义 CA 抑制剂的抗病毒作用，以促进开发基于 CA 的新型抗病毒疗法（NIH 的高优先级 HIV/AIDS 研究领域）。