HIV-1 Replication Without Integration

HIV-1 复制无需整合

基本信息

批准号：
8679470
负责人：
DAVID N LEVY
金额：
$ 22.21万
依托单位：
NEW YORK UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-19 至 2015-08-18
项目状态：
已结题

项目摘要

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.

HIV-1感染的持久性是许多因素的结果，包括快速的病毒进化，逃避免疫力，建立潜在和神秘地复制病毒的储层以及对免疫系统的损害直接或直接由病毒复制引起。我们对这些因素的理解仍然不足以完全解释HIV-1的持久性和发病机理。提出的研究将描述以前未知的HIV-1复制途径，我们发现可能有助于HIV-1适应，建立病毒储存库和发病机理。作为逆转录病毒，将HIV-1 DNA整合到细胞染色体中是生产性感染所必需的。有趣的是，体内和体外的HIV-1 DNA中有90-99％保持不整合，并且本身无法产生足够的RNA和蛋白质来制造新的病毒体。但是，我们已发表的研究表明，在一个有效的感染细胞中，乌德纳（UDNA）与综合病毒相辅相成，并完成了其复制周期。换句话说，UDNA有助于复制的病毒群体，并放大多种感染的量。结果是增加有效的病毒种群大小，可能发散病毒之间的丰富相互作用，并通过重组和突变增强了病毒的进化。 HIV-1复制的这种新型机制与前整合潜伏期不同，不取决于乌德纳的后续整合。新的初步数据表明，HIV-1可以超级感染细胞并绕过整合，从而导致病毒复制加速，这是病毒适应性的重要参数。我们假设UDNA对病毒进化，持久性和发病机理产生了强烈的影响。由于非增殖细胞中乌德纳的圆形形式的高稳定性，我们假设UDNA构成了长期寿命的病毒储存库，其复制是通过对宿主细胞的生产性恢复来恢复的。通过实验和分析方法的结合，以下特定目标将检验这些假设。目标1。乌德纳对复制病毒人群的贡献是什么？ AIM 2。检验udna可以作为T细胞和巨噬细胞中潜在病毒的储层的假设。目的3。检查紫外线对HIV-1复制动力学的影响。目标4。开发乌德纳对HIV-1复制和多样化的贡献的描述性和预测性数学模型，以探讨我们的主要假设。