HIV-1 Replication Without Integration

HIV-1 复制无需整合

• 批准号: 8432850
• 负责人: DAVID N LEVY
• 金额: $ 34.82万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2016-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8432850
• 关键词: Antiviral Agents; Archives; Binding; Bypass; Cell Culture Techniques; Cell Survival; Cells; Chromosomes; Collaborations; Complement; Complex; DNA; Data; Event; Evolution; Gene Expression; Gene Pool; Genetic Recombination; Genetic Transcription; Genome; HIV-1; Health; Immune system; Immunity; In Vitro; Infection; Junk DNA; Kinetics; Latent Virus; Life; Modeling; Mutation; Natural History; Nuclear Export; Output; Pathogenesis; Pathway interactions; Population; Population Sizes; Production; Proliferating; Proteins; Proviruses; Publishing; RNA; Rest; Retroviridae; Role; T-Lymphocyte; Testing; Transcript; Vaccines; Variant; Viral; Viral Vector; Virion; Virus; Virus Diseases; Virus Replication; Waste Products; fitness; improved; in vivo; macrophage; mathematical model; novel; predictive modeling; superinfection; therapy development; transmission process; vaccine development; virus genetics

DESCRIPTION (provided by applicant): The persistence of HIV-1 infection is the result of many factors, including rapid viral evolution to evade immunity, the establishment of reservoirs of both latent and cryptically replicating virus, and damage to the immune system caused directly or indirectly by virus replication. Our understanding of each of these factors remains inadequate to fully explain HIV-1 persistence and pathogenesis. The studies proposed will describe a previously unknown pathway of HIV-1 replication we have discovered that likely contributes to HIV-1 adaptation, to the establishment of viral reservoirs, and to pathogenesis. As a retrovirus, integration of HIV-1 DNA into the cellular chromosome is necessary for productive infection. Interestingly, 90-99% of HIV-1 DNA in vivo and in vitro remains unintegrated and by itself is unable to generate sufficient RNA and proteins to make new virions. However, our published studies reveal that in a productively infected cell, uDNA is complemented by the integrated provirus and completes its replication cycle. In other words, uDNA contributes to the replicating virus population and magnifies the amount of multiple infection. The result is an increased effective virus population size, abundant interactions among potentially divergent viruses, and enhanced virus evolution through recombination and mutation. This novel mechanism for HIV-1 replication is distinct from pre-integration latency and does not depend on subsequent integration by the uDNA. New preliminary data indicate the HIV-1 can superinfect cells and bypass integration, resulting in accelerated viral replication, an important parameter of viral fitness. We hypothesize that uDNA exerts a strong influence on viral evolution, persistence and pathogenesis. Owing to the high stability of circular forms of uDNA in non-proliferating cells, we hypothesize that uDNA constitutes a long-lived reservoir of virus whose replication is restored by productive reinfection of the host cell. Through a combination of experimental and analytical approaches, the following specific aims will test these hypotheses. Aim 1. What is the contribution of uDNA to the replicating virus population? Aim 2. Test the hypothesis that uDNA can function as a reservoir of latent viruses in T cells and macrophages. Aim 3. Examine the influence of uDNA on HIV-1 replication kinetics. Aim 4. Develop descriptive and predictive mathematical models of uDNA's contribution to HIV-1 replication and diversification to explore our main hypothesis.

描述（申请人提供）：HIV-1感染的持续存在是多种因素共同作用的结果，包括病毒快速进化以逃避免疫、潜伏病毒和隐性复制病毒储存库的建立，以及病毒复制直接或间接对免疫系统造成的损害。我们对这些因素的了解仍然不足以充分解释 HIV-1 的持续存在和发病机制。拟议的研究将描述一种以前未知的 HIV-1 复制途径，我们发现该途径可能有助于 HIV-1 适应、病毒储存库的建立和发病机制。作为一种逆转录病毒，HIV-1 DNA 整合到细胞染色体中对于有效感染是必要的。有趣的是，90-99%的HIV-1 DNA在体内和体外仍未整合，其本身无法产生足够的RNA和蛋白质来制造新病毒体。然而，我们发表的研究表明，在有效感染的细胞中，uDNA 被整合的原病毒补充并完成其复制周期。换句话说，uDNA有助于病毒种群的复制，并放大多重感染的数量。其结果是有效病毒种群规模的增加、潜在分歧病毒之间的丰富相互作用以及通过重组和突变增强病毒进化。这种 HIV-1 复制的新机制与整合前潜伏期不同，并且不依赖于 uDNA 的后续整合。新的初步数据表明，HIV-1 可以重复感染细胞并绕过整合，从而加速病毒复制，这是病毒适应性的一个重要参数。我们假设 uDNA 对病毒进化、持久性和发病机制产生强烈影响。由于uDNA的环状形式在非增殖细胞中具有高度稳定性，我们假设uDNA构成了一个长寿命的病毒库，其复制可通过宿主细胞的生产性再感染而恢复。通过实验和分析方法的结合，以下具体目标将检验这些假设。目标 1. uDNA 对复制病毒群体有何贡献？目标 2. 检验 uDNA 可以作为 T 细胞和巨噬细胞中潜伏病毒库的假设。目标 3. 检查 uDNA 对 HIV-1 复制动力学的影响。目标 4. 开发 uDNA 对 HIV-1 复制和多样化贡献的描述性和预测性数学模型，以探索我们的主要假设。