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

基本信息

批准号：
10247079
负责人：
Deborah Persaud
金额：
$ 76.99万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
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/10247079
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 base 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 potential biomarker prevent residence response sex single-cell RNA sequencing therapeutic target transcriptome transcriptomics viral rebound virology

项目摘要

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.

全球近 3700 万艾滋病毒感染者中的潜伏艾滋病毒无法治愈，其中 170 万是艾滋病毒感染者孩子们。消除潜伏病毒库 (LR) 对于无抗逆转录病毒治疗 (ART) 的缓解至关重要，其中停止 ART 后不会发生病毒反弹。潜伏期逆转剂 (LRA) 可以治疗靶向 LR 并使其容易被消除。成人 LRA 的临床试验正在进行中并计划进行围产期感染的儿童。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（通过围产期感染），逆转速度较慢且幅度较小与成年时期相比。我们的研究结果表明，这种主要的治疗方法需要进一步的研究体内研究破译围产期感染中 HIV 潜伏期逆转的机制，以指导临床在这一人群中进行上帝抵抗军试验。我们假设 LR 建立的免疫环境并且存在于围产期感染中，使其本质上比成人感染更能抵抗潜伏期逆转。该应用程序的具体目标是： 1) 确定并比较尺寸、成分和围产期和成人感染中潜伏艾滋病毒储存库的可诱导性，并描述它们的差异； 2) 通过 CD4+ 转录组分析确定前病毒再激活易感性的相关性 T 细胞在围产期和成人感染中的作用； 3) 定义调节性 T 细胞 (Treg) 对围产期 HIV 感染中的潜在 HIV 储存库并探索单细胞 RNA-seq 方法的实用性检查 CD4+ T 细胞亚群对潜伏期逆转的差异反应。我们将招募围产期艾滋病毒感染者在美国和乌干达照顾的受感染儿童、青少年和成人，并全面描述并比较潜伏病毒库的大小，通过总和完整的原病毒 DNA（包括一体化）。我们将确定最大 T 细胞激活条件下对潜伏期逆转的敏感性，以及临床相关的潜伏期逆转疗法（TLR-7 激动剂 GS-9620、IL-15 超级激动剂 N-803 或 SMAC 模拟物-AZD5582），并按感染模式、LRA 类别、病毒学持续时间进行分析抑制、原病毒载量和亚型。免疫激活基线状态之间的相关性，以及 CD4+ T 细胞的基线转录组作为感染模式、地理区域/HIV 亚型的函数，以及诱导水库的大小将被确定。我们将进一步研究 Tregs 和非 Tregs 对围产期感染中的 LR，通过单细胞 RNA-seq 的探索性研究来定义基线转录不同 CD4+ 记忆 T 细胞亚群（包括 Tregs）的概况，以及它们对远程监视系统。这里提出的系统特征将为围产期艾滋病毒潜伏期的机制见解提供信息，包括调节性 T 细胞 (Treg) 的贡献，并提供有关 LRA 在围产期应用的关键数据感染，以及用于测量 LRA 在该人群中的功效的最佳生物标志物。