Latent Viral Reservoirs in HIV 1 Infection

描述(由申请人提供)：HIV-1在静止的CD4+T细胞中的潜在储存库是根除病毒的主要障碍。目前，人们对消除这种储藏库的潜在策略非常感兴趣。在之前的资助期间，我们进行了机制研究，导致了对当前潜伏期模型的重新评估，并表明潜伏期机制在原代细胞系统中得到了最准确的分析，而不是在转化后的T细胞系中。我们开发了一种体外系统，在该系统中可以产生携带潜伏的HIV-1基因组的初级静止CD4+T细胞。在这项提案中，我们描述了使用这一系统来识别和表征能够重新激活潜伏的HIV-1的小分子，以用于根除这种宿主的努力。我们已经确定了一个有希望的热门作品，我们将对其进行机械描述。此外，我们将使用这一系统来探索有关潜在储存库动态的关键悬而未决的问题，重点是对根除战略至关重要的问题。其具体目的是(1)确定已确定的HIT(5-羟基-1，4-萘醌)重新激活潜伏HIV-1的机制，(2)利用原代静止CD4+T细胞中新型的潜伏期模型来筛选其他在不诱导全球T细胞激活的情况下重新激活潜伏的HIV-1的药物，(3)对重新激活潜伏的HIV-1的药物的体外疗效进行比较评估，以及(4)使用原代细胞模型来探索维持HIV-1潜伏期的机制。与公共卫生相关：即使在接受有效抗病毒药物治疗的患者中，艾滋病毒也可能以沉默或潜伏的形式持续存在。治愈艾滋病毒感染将需要新的方法来消除这种潜在的蓄水池。这里提出的研究旨在寻找可以消除这种潜在储存库的新药。

DESCRIPTION (provided by applicant): The latent reservoir for HIV-1 in resting CD4+ T cells is a major barrier to virus eradication. There is great current interest in potential strategies for eliminating this reservoir. During the previous funding period, we have carried out mechanistic studies that have led to a reevaluation of current models of latency and have shown that mechanisms of latency are most accurately analyzed in primary cell systems rather than in transformed T cell lines. We developed an in vitro system in which primary resting CD4+ T cells carrying latent HIV-1 genomes can be generated. In this proposal, we describe the use of this system to identify and characterize small molecules capable of reactivating latent HIV-1 for use in efforts to eradicate this reservoir. We have already identified one promising hit which we will characterize mechanistically. In addition, we will use this system to explore key unanswered questions regarding the dynamics of the latent reservoir, focusing on issues that are important for eradication strategies. The Specific Aims are (1) to determine the mechanism by which the identified hit (5-hydroxy-1,4-naphthoquinone) reactivates latent HIV-1, (2) to use the novel latency model in primary resting CD4+ T cells to screen selected small molecule libraries for other agents that reactivate latent HIV-1 without inducing global T cell activation, (3) to carry out a comparative in vitro evaluation of the efficacy of agents that reactivate latent HIV-1, and (4) to use the primary cell model to explore mechanisms involved in the maintenance of HIV-1 latency. PUBLIC HEALTH RELEVANCE: HIV can persist in a silent or latent form even in patients who are treated with potent antiviral drugs. Curing HIV infection will require new ways to eliminate this latent reservoir. The studies proposed here are designed to find new drugs that can eliminate this latent reservoir.