Deciphering Mechanisms of HIV Latency Reversal in Perinatal Infections

潜伏的艾滋病毒阻碍了全世界近3700万艾滋病毒携带者的治愈，其中170万人是 孩子们。消除潜伏期(LR)对于抗逆转录病毒疗法(ART)的无缓解至关重要，其中 当ART停止时，病毒反弹不会发生。潜伏期反转剂(LRA)可以在治疗上靶向 并使其容易被消除。成人LRA的临床试验正在进行中，并计划于#年进行。 围产期感染的儿童。然而，关键的是，我们最近的体外研究表明，潜伏期的动力学 当LR在婴儿期(通过围产期感染)建立时，逆转较慢且幅度较小 与成年期相比。我们的发现表明，这种主要的治疗方法需要进一步的治疗。 活体研究破译围产期感染HIV潜伏期逆转的机制以指导临床 在这群人中进行LRA试验。我们假设建立LR的免疫环境 并存在于围产期感染中，使其对潜伏期逆转的抵抗力天生强于成人感染。 本申请的具体目的是：1)确定和比较尺寸、组成和 围产期和成人感染中潜伏的艾滋病毒蓄积物的诱导性，并表征它们的差异； 2)通过转录分析确定前病毒重新激活的易感性 T细胞在围产期和成人感染中的作用；以及3)确定调节性T细胞(Tregs)对 围产期HIV感染中潜伏的HIV蓄积及单细胞RNA-SEQ方法的应用探讨 检测CD4T细胞亚群对潜伏期逆转的不同反应。我们将招收围产期艾滋病毒感染者- 受感染的儿童、青少年和成人在美国和乌干达得到照顾，并全面描述 并比较以总的前病毒DNA和完整的前病毒DNA(包括 整合)。我们将确定在最大T细胞激活条件下对潜伏期逆转的敏感性 临床相关潜伏期逆转疗法(TLR-7激动剂GS-9620、IL-15超级激动剂N-803或 Smac模拟物-AZD5582)，当通过感染模式、LRA分类、病毒学持续时间进行分析时 抑制、前病毒负荷和亚型。免疫激活的基线状态之间的相关性，以及 CD4T细胞的基线转录水平与感染模式、地理区域/HIV亚型以及 诱发水库的大小将被确定。我们将进一步研究Tregs和Non-Tregs对 围产期感染中的LR与单细胞RNA-SEQ在确定基线转录水平中的探索性研究 不同的CD4记忆T细胞亚群，包括树突状细胞，以及它们对 LRAS。这里提出的系统描述将为对围产期艾滋病毒潜伏期的机械性洞察提供信息， 包括调节性T细胞(Tregs)的贡献，并提供LRAs在围产期应用的关键数据 感染，以及衡量LRA在这一人群中有效性的最佳生物标记物。

Latent HIV prevents cure for the nearly 37 million persons living with HIV worldwide, of whom 1.7 million are children. Elimination of the latent reservoir (LR) is critical for antiretroviral therapy (ART)-free remission, where viral rebound does not occur when ART is stopped. Latency reversal agents (LRAs) can therapeutically target the LR and render it susceptible to elimination. Clinical trials of LRAs in adults are ongoing and planned for perinatally-infected children. Critically, however, our recent in vitro studies reveal that the kinetics of latency reversal are slower and of lower magnitude when the LR is established in infancy (through perinatal infection) compared with during adulthood. Our findings suggest that this major therapeutic approach requires further ex vivo studies to decipher mechanisms of HIV latency reversal in perinatal infections in order to guide clinical trials of LRAs in this population. We hypothesize that the immune environment in which the LR is established and exists in perinatal infection renders it intrinsically more resistant to latency reversal than in adult infection. The specific aims of this application are: 1) Determine and compare the size, composition, and inducibility of the latent HIV reservoir in perinatal and adult infection, and characterize their differences; 2) Identify correlates of susceptibility to proviral reactivation through transcriptomic analyses of CD4+ T cells in perinatal and adult infections; and 3) Define the contribution of regulatory T cells (Tregs) to the latent HIV reservoir in perinatal HIV infection and explore the utility of single-cell RNA-seq approaches to examine differential responses of CD4+ T cell subsets to latency reversal. We will enroll perinatally HIV- infected children, adolescents, and adults cared for in the US and Uganda, and comprehensively characterize and compare the size of the latent reservoir, as measured by total and intact proviral DNA (including sites of integration). We will determine susceptibility to latency reversal under maximum T cell activating conditions and clinically relevant latency reversal therapeutics (TLR-7 agonist GS-9620, the IL-15 superagonist N-803, or the SMAC mimetic-AZD5582), and when analyzed by mode of infection, LRA class, duration of virologic suppression, proviral load, and subtype. Correlations between baseline states of immune activation, along with baseline transcriptomes of CD4+ T cells as a function of mode of infection, geographic region/HIV subtype, and size of the induced reservoir will be determined. We will further examine contribution of Tregs and non-Tregs to the LR in perinatal infections, with exploratory studies of single-cell RNA-seq in defining baseline transcriptional profiles of the different CD4+ memory T cell subsets, including Tregs, and their differential responses to the LRAS. The systematic characterization proposed here will inform mechanistic insights into perinatal HIV latency, including the contribution of regulatory T cells (Tregs), and provide critical data on the utility of LRAs in perinatal infections, along with optimal biomarkers for measuring efficacy of LRAs in this population.