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 消除整合的，复制能力的艾滋病毒前病毒从宿主基因组持续存在，尽管 抑制性抗逆转录病毒疗法（ART）是功能性治愈的主要障碍。细胞携带这些 前病毒类型产生边缘水平的病毒产物， 监督机制。这种缺乏检测的免疫系统，除了其增加的增长 由于稳态增殖和克隆扩增，潜在的延长潜伏感染细胞的寿命 从而产生持久储集层。人们对精确治疗的潜力抱有极大的热情 在临床环境中瞄准潜在的储库。为了实现这一重大的生物医学目标，我们必须首先发现 细胞内在和/或外在“介质”的水库持久性和潜伏期重新激活开关之前，我们可以 利用这些知识进行临床干预。为了填补这一知识空白，我们实施了一项增益- 在含有潜伏原病毒的CD 4 + T细胞系中进行功能筛选，发现了几种新的调节因子， 潜伏激活开关在这个探索性和发展性的R21赠款提案中，我们将重点关注我们的 其中一种激活剂ADAP 1（Arf-GAP with dual PH domain-containing protein-1）。 重要的是，ADAP 1以前没有与免疫细胞生物学或HIV前病毒的控制相关， 基因调控和潜伏期的关系，从而使我们的研究具有概念上的创新。 鉴于这些发现，本研究建议的主要目的是更好地了解 ADAP 1控制HIV潜伏期和再激活的分子机制。为了做到这一点，我们 将在一组全面的CD 4 + T细胞效应子亚群中利用主要潜伏期模型的能力， 了解复杂和异质性的免疫细胞辅因子-HIV前病毒基因组相互作用， 潜伏期和再激活。该建议的中心假设是ADAP 1是激活细胞所必需的。 CD 4 + T细胞亚群中的信号传导-转录调节程序，而静息细胞暴露于免疫 细胞刺激/共刺激。具体来说，我们将探索ADAP 1是否能从HIV感染者中重新激活潜伏的HIV。 各种CD 4 + T细胞亚群的潜伏期模型（Aim 1），并询问ADAP 1调节的细胞信号传导- 转录程序导致潜伏的HIV再活化（Aim 2）。这些研究可能会带来革命性的 影响我们对HIV潜伏生物学的理解，并具有治疗意义。通过阐明如何 CD 4 + T细胞亚群塑造感染过程，以及HIV如何为感染者选择宿主资源（如ADAP 1）。 潜伏期-再激活开关，我们将深入了解基本过程以及可以靶向的途径 帮助实现NIAID结束艾滋病流行的使命。