Enhanced latency reversal and reservoir clearance in macaques

项目摘要 目前对艾滋病毒感染的治疗需要严格遵守每日抗逆转录病毒治疗（ART）。艺术 大大降低了死亡率和发病率，但不能治愈。治愈的主要障碍是潜在的 感染的CD 4 + T细胞携带整合的HIV前病毒，能够重新激活并引起 如果停止抗逆转录病毒疗法，病毒血症复发。一个主要的战略，以消除这一持久性水库被称为 “休克和杀死”或“踢和杀死”，并包括两个干预与持续的艺术：第一，潜伏期 逆转剂（LRA），以重新激活潜伏病毒和第二，一种方法，以提高感染细胞的清除。 在这个建议中，我们将建立在我们的初步数据表明，激活的非典型（nc）NF-κ B-κ B的， κB信号通路与第二种来源的半胱天冬酶激活剂（SMACm）的模拟物 导致动物模型中的全身潜伏期逆转。使用高度相关的SIV/RM/ART模型，我们将 回答了关于SMACm治疗的生物学效应的三个重要问题。首先（目标1），我们将 确定ART期间SMACm治疗后产生的病毒血症的解剖学和细胞起源 与ART中断后病毒血症反弹的关系。在这项研究中，我们将利用条形码 SIVmac 239 M在来自血液和组织的分选的CD 4 + T细胞亚群中广泛测序SIV RNA， 与血浆病毒相比。我们还将应用单细胞转录组学方法来回答这个问题 问题第二（目标2），我们测试了一种新的“踢和杀”的方法，结合延迟逆转与SMACm 加上维奈托克，一种选择性BCL-2抑制剂，已被证明可引起HIV-1细胞的优先凋亡。 表达细胞。我们将确定这种联合治疗是否减少总的，完整的，和复制 在来自多种组织的记忆性CD 4 + T细胞中有能力的储库和/或调节病毒复发后 ART中断。第三（目标3），我们将对预测病毒再活化的免疫病毒学因素进行建模， ART联合SMACm治疗以及在ncNF-κB治疗靶向后ART中断期间， BCL-2通路。该模型将进一步揭示这种治疗方法的潜力，并为设计提供信息。 未来的“踢死”研究。 这项工作将使我们能够阐明靶向ncNF-κB通路的潜伏期逆转的全部潜力， 评估减少病毒持久性的创新策略。我们希望这些结果将有助于治愈 艾滋病病毒感染。

PROJECT SUMMARY Current treatment for HIV infection requires strict adherence to daily antiretroviral therapy (ART). ART has greatly reduced mortality and morbidity but is not a cure. The main barrier to a cure is the pool of latently infected CD4+ T cells that carry integrated HIV provirus and are capable of reactivating and causing recrudescence of viremia if ART is stopped. A leading strategy to eliminate this persistent reservoir is termed “shock and kill” or “kick and kill” and consists of two interventions used with continued ART: first, a latency reversal agent (LRA) to reactivate latent virus and second, an approach to enhance clearance of infected cells. In this proposal, we will build upon our preliminary data indicating that activation of the non-canonical (nc) NF- κB signaling pathway with a mimetic of the second mitochondrial-derived activator of caspases (SMACm) results in systemic latency reversal in animal models. Using the highly relevant SIV/RM/ART model, we will answer three important questions regarding the biologic effects of SMACm treatment. First (Aim 1), we will determine the anatomic and cellular origin of the viremia produced during ART upon treatment with SMACm and its relationship with rebound viremia after ART interruption. In this study, we will utilize barcoded SIVmac239M to extensively sequence SIV RNA in sorted CD4+ T cell subsets from blood and tissues in comparison to plasma virus. We will also apply single cell transcriptomic methodologies to answer this question. Second (Aim 2), we test a novel “kick and kill” approach that combines latency reversal with SMACm plus venetoclax, a selective BCL-2 inhibitor, that has been shown to cause preferential apoptosis of HIV- expressing cells. We will determine whether this combined treatment reduces total, intact, and replication competent reservoirs in memory CD4+ T cells from multiple tissues and/or modulates viral recrudescence after ART interruption. Third (Aim 3), we will model the immunovirologic factors predictive of virus reactivation on ART with SMACm treatment and during ART interruption following therapeutic targeting of the ncNF-κB and BCL-2 pathways. This model will further reveal the potential of this cure approach and inform the design of future “kick and kill” studies. This work will allow us to elucidate the full potential of targeting the ncNF-κB pathway for latency reversal and to evaluate an innovative strategy to reduce virus persistence. We hope these results will contribute to a cure for HIV infection.