Deciphering Mechanisms of HIV Latency Reversal in Perinatal Infections

破译围产期感染中 HIV 潜伏期逆转的机制

• 批准号: 10686028
• 负责人: Deborah Persaud
• 金额: $ 75.04万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10686028
• 关键词: Adolescent; Adult; Agonist; Biological Markers; CD4 Positive T Lymphocytes; Caring; Cell Compartmentation; Cells; Child; Childhood; Clinical Trials; Clinical Trials Design; Clonal Expansion; DNA; Data; Disease remission; Enrollment; Environment; Exposure to; Gene Expression Profile; Gene Silencing; Genes; Genetic; Genomics; Geographic Locations; HIV; HIV Infections; HLA-DR Antigens; IL2RA gene; Immune; In Vitro; Infection; Interleukin-15; Kinetics; Knowledge; Location; Measures; Participant; Perinatal; Perinatal Infection; Persons; Population; Predisposition; Regulation; Regulatory T-Lymphocyte; Resistance; Resolution; Rest; Sampling; Stimulus; T-Cell Activation; T-Lymphocyte; T-Lymphocyte Subsets; Therapeutic; Uganda; Viral; Virus; antiretroviral therapy; biomarker discovery; clinically relevant; cytokine; defined contribution; exhaustion; immune activation; infancy; insight; integration site; latent HIV reservoir; memory CD4 T lymphocyte; mimetics; novel; perinatal HIV; permissiveness; potential biomarker; prevent; residence; response; sex; single-cell RNA sequencing; therapeutic target; transcriptome; transcriptomics; viral rebound

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.

潜在的艾滋病毒可防止近3700万艾滋病毒的人治愈，其中170万 孩子们。消除潜在储层（LR）对于抗逆转录病毒疗法（ART） - 无缓解至关重要，其中 停止艺术时不会发生病毒反弹。潜伏期逆转剂（LRAS）可以治疗靶向 LR和使其容易消除。成人LRA的临床试验正在进行中，并计划 围产期感染的儿童。然而，至关重要的是，我们最近的体外研究表明，潜伏期的动力学 当LR在婴儿期建立时，逆转较慢，幅度较低（通过围产期感染） 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 为了引导临床 LRA在该人群中的试验。我们假设建立LR的免疫环境 并且存在于围产期感染中，其本质上比在成人感染中更能抵抗潜伏期逆转。 本应用程序的具体目的是：1）确定和比较大小，组成和 潜在的HIV储藏在围产期和成人感染中的诱导性，并表征其差异； 2）通过CD4+的转录组分析确定对病毒重新激活的敏感性的相关性 围产期和成人感染的T细胞； 3）定义调节T细胞（Treg）对 围产期HIV感染中潜在的HIV水库并探索单细胞RNA-SEQ方法的实用性 检查CD4+ T细胞子集对潜伏期逆转的差异响应。我们将在周围注册HIV- 受感染的儿童，青少年和在美国和乌干达受到照顾的成年人，并全面地描述了 并比较通过总和病毒DNA测量的潜在储层的大小（包括 一体化）。我们将在最大T细胞激活条件下确定对潜伏期逆转的敏感性和 临床上相关的潜伏期逆转疗法（TLR-7激动剂GS-9620，IL-15超级机动器N-803或 Smac Mimetic-Azd5582），当通过感染模式分析时，LRA类，病毒学持续时间 抑制，前病毒载荷和亚型。免疫激活基线状态之间的相关性以及 CD4+ T细胞的基线转录组是感染模式，地理区域/HIV亚型和 将确定诱导储层的大小。我们将进一步研究Treg和非Tregs对 围产期感染中的LR，对单细胞RNA-Seq的探索性研究定义基线转录 不同CD4+存储T细胞子集的概况，包括Treg及其对差异响应 lras。这里提出的系统表征将为围产期艾滋病毒潜伏期的机理见解提供信息， 包括调节T细胞（TREG）的贡献，并提供有关LRA在围产期效用的关键数据 感染，以及用于测量LRA在该人群中的功效的最佳生物标志物。