Exploring the mechanism of ADAP1 control of HIV latency and reactivation in CD4 T cells

探索 ADAP1 控制 CD4 T 细胞中 HIV 潜伏期和再激活的机制

• 批准号: 10082399
• 负责人: Ivan D'Orso
• 金额: $ 24.51万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10082399
• 关键词: Agonist; Applications Grants; Attention; Binding; Biology; CD4 Positive T Lymphocytes; Cell Line; Cell Shape; Cell membrane; Cells; Cellular biology; Clinical; Clonal Expansion; Complex; Detection; Development; Disease Progression; Effector Cell; Epidemic; Event; Exposure to; GTPase-Activating Proteins; Gene Expression Regulation; Genetic Screening; Genetic Transcription; Genome; Goals; Growth; HIV; Immune; Immune system; Immunization; Immunologic Surveillance; Impairment; Infection; Interphase Cell; Intervention; KRAS2 gene; Knowledge; Lead; Link; Longevity; Lymphocyte; Maintenance; Mediating; Mediator of activation protein; Mission; Modeling; Molecular; Names; National Institute of Allergy and Infectious Disease; Neuraxis; PH Domain; Pathway interactions; Patients; Phosphotransferases; Physiological; Precision therapeutics; Process; Proteins; Proviruses; Refractory; Regulation; Regulatory T-Lymphocyte; Research Proposals; Resources; Rest; Sampling; Shapes; Signal Transduction; System; T memory cell; T-Cell Activation; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Therapeutic; Transcriptional Activation; Viral; Work; antiretroviral therapy; clinically relevant; effector T cell; gain of function; innovation; insight; memory CD4 T lymphocyte; novel; programs; promoter; ras GTPase-Activating Proteins; reactivation from latency; response; targeted treatment; therapeutic target; transcription factor; virtual

PROJECT SUMMARY Elimination of integrated, replication-competent HIV proviruses from host genomes persisting despite suppressive anti-retroviral therapy (ART) is the major roadblock to a functional cure. Cells harboring these types of proviruses produce marginal levels of viral products thereby becoming refractory to immune surveillance mechanisms. This lack of detection by the immune system, in addition to its increased growth potential, due to homeostatic proliferation and clonal expansion, extend the lifespan of latently infected cells generating a persistent reservoir. There is enormous enthusiasm for the potential of precision therapies targeting the latent reservoir in clinical settings. To achieve this major biomedical goal, we must first discover cell-intrinsic and/or -extrinsic “mediators” of reservoir persistence and latency-reactivation switch before we can leverage this knowledge for clinical intervention. To start filling this gap in knowledge, we implemented a gain- of-function screen in a CD4+ T cell line containing a latent provirus and discovered several novel regulators of the latency-reactivation switch. In this exploratory and developmental R21 grant proposal, we will focus our attention on one of those activators named ADAP1 (Arf-GAP with dual PH domain-containing protein-1). Importantly, ADAP1 has not been previously linked to immune cell biology nor to the control of HIV proviral gene regulation and latency, thus our studies hold conceptual innovation. Given these findings, the major objective of this research proposal is to achieve a better understanding of the molecular mechanisms underlying ADAP1 control of HIV latency and reactivation. To accomplish this, we will leverage the power of primary models of latency in a comprehensive set of CD4+ T cell effector subsets to understand complex and heterogeneous immune cell co-factor–HIV proviral genome interactions during latency and reactivation. The central hypothesis of this proposal is that ADAP1 is required to activate cell signaling-transcription regulatory programs in CD4+ T cell subsets while resting cells are exposed to immune cell stimulation/co-stimulation. Specifically, we will explore if ADAP1 functions to reactivate latent HIV from latency models in various CD4+ T cell subsets (Aim 1), and interrogate the ADAP1-regulated cell signaling- transcriptional programs leading to latent HIV reactivation (Aim 2). These studies could have a revolutionary impact on our understanding of HIV latency biology and have therapeutic implications. By elucidating how CD4+ T cell subsets shape the course of infection and how HIV co-opts host resources (like ADAP1) for the latency-reactivation switch, we will gain insights into basic processes as well as pathways that can be targeted therapeutically to help achieve NIAID mission’s of ending the HIV epidemic.

项目摘要 消除从宿主基因组中持续存在的综合，复制能力的HIV Proviruse 抑制性抗逆转录病毒疗法（ART）是功能治愈的主要障碍。带有这些的细胞 预科病毒的类型产生了边缘水平的病毒产品，从而变得难治性 监视机制。免疫系统缺乏检测，除了增长的增长 由于稳态增殖和克隆膨胀，潜力延长了潜在感染细胞的寿命 产生持续的水库。对精确疗法的潜力充满热情 针对临床环境中的潜在储层。为了实现这一主要的生物医学目标，我们必须首先发现 在我们可以之前 利用这些知识进行临床干预。为了开始填补这一知识的差距，我们实施了收益 - CD4+ T细胞系中包含潜在病毒的功能屏幕，发现了几个新的调节剂 延迟反应开关。在这个探索性和发展性R21赠款提案中，我们将集中精力 注意一个名为ADAP1的激活剂（具有双pH结构域蛋白1）的激活剂。 重要的是，ADAP1先前尚未与免疫细胞生物学相关，也不与HIV病毒的控制有关 基因调节和潜伏期，因此我们的研究具有概念创新。 鉴于这些发现，该研究建议的主要目标是更好地理解 ADAP1控制HIV潜伏期和重新激活的分子机制。为此，我们 将在一组CD4+ T细胞效应子集中利用潜伏期的主要模型的力量 了解复合物和异构免疫核物共同因素 - HIV前病毒基因组相互作用 潜伏期和重新激活。该提案的中心假设是激活细胞需要ADAP1 CD4+ T细胞子集中的信号传导转录调节程序，而静止细胞暴露于免疫 细胞刺激/共刺激。具体来说，我们将探讨ADAP1是否功能使潜在的HIV重新激活 各种CD4+ T细胞子集中的延迟模型（AIM 1），并询问ADAP1调节的细胞信号 - 转录程序导致潜在的HIV重新激活（AIM 2）。这些研究可能具有革命性 影响我们对艾滋病毒潜伏生物学的理解并具有治疗意义。通过阐明如何 CD4+ T细胞子集塑造感染过程以及HIV合作如何托管资源（如ADAP1） 延迟反应开关，我们将获得对基本过程以及可以针对的途径的见解 在治疗上，有助于实现NIAID任务结束艾滋病毒流行。