Overcoming HIV-1 transcriptional latency in unresponsive CD4 T cells

克服无反应 CD4 T 细胞中的 HIV-1 转录潜伏期

• 批准号: 9410387
• 负责人: OLAF KUTSCH
• 金额: $ 66.26万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9410387
• 关键词: AIDS therapy; ATAC-seq; Address; Affect; BACH2 gene; Binding; Biological Markers; CD28 gene; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cell Differentiation process; Cell Line; Cell physiology; Cell surface; Cells; Characteristics; Complex; Data; Development; Devices; Drug Combinations; Drug Compounding; Drug Targeting; Event; Exhibits; Frequencies; Future; Gene Expression; Genetic Transcription; Goals; HIV; HIV Infections; HIV-1; Infection; Intervention; Investigation; Link; Mature T-Lymphocyte; Memory; Molecular; Molecular Biology; Molecular Target; PPM1D gene; Patients; Pattern; Peroxisome Proliferator-Activated Receptors; Pharmacology; Phenotype; Phosphotransferases; Population; Proliferating; Reporting; Research; Resistance; Signal Transduction Pathway; System; Systems Biology; T memory cell; T-Lymphocyte; TCR Activation; Testing; Therapeutic; Viral reservoir; Virus; Virus-Cell Membrane Interaction; base; design; genetic manipulation; in vitro Model; in vivo; innovation; latent infection; molecular phenotype; multicatalytic endopeptidase complex; new therapeutic target; novel; prevent; reconstitution; restoration; small hairpin RNA; specific biomarkers; transcription factor

ABSTRACT Latent HIV-1 infection has been recognized as a major obstacle to the development of a curative HIV-1 therapy, but host cell-virus interactions that control latent infection are still ill defined. Key to this application is the realization that the host cells of latent HIV-1 infection events are actually phenotypically altered in a manner that (i) forces the virus into a latent state and that (ii) renders the host cells unresponsive to stimulation, thereby preventing efficient therapeutic induction of HIV-1 reactivation. These changes go beyond a simple quiescent state that is characteristic for functional memory T cells. This proposal will extend on these findings and seek to address three major roadblocks in the field of HIV latency research. Roadblock #1 (Aim 1) concerns our inability to identify biomarkers that specifically define T cell sub-populations in which latently infected T cells are highly enriched. Such biomarkers would allow us to detail the molecular biology of the host cell state that enables and maintains latent HIV-1 infection. Leading to this application, we found that the intracellular changes allowing latent HIV-1 infected to persist are associated with a unique CD4+CD28– CD9+CD151+ phenotype, a T cell phenotype that also demarcates a small CD4 T cell sub-population that is increased in HIV patients on ART. We already demonstrate that CD151 expression by itself is associated with reservoir capacity. We will now detail the viral reservoir capacity of T cell sub-populations described by CD28, CD151 and CD9 expression. Roadblock #2 (Aim 2) addressed in this application is the question how the intracellular changes observed in host cells of latent HIV infection events act (i) to control HIV transcription and and (ii) to suppress T cell responsiveness. In addition to the proposed research on host-cell factors/networks controlling latent HIV infection, we already have identified several targets/interaction networks that control latent infection and that will be immediately probed. We will further address roadblock #3 (Aim 3), the question why stimulation of ex vivo T cell material from HIV patients seems to only trigger HIV-1 reactivation in a small fraction of the latently HIV-infected cells. Based on our results, we predict that T cell subpopulations described by differential CD28, CD9 and CD151 expression patterns, will exhibit varying levels of unresponsiveness to stimulation, allowing latent HIV infection to persist with different efficacies. Based on the results we have obtained leading to this application and the additional results we will generate, we will immediately begin to rationally design compound-based intervention strategies that would (i) first reconstitute T cell responsiveness in reservoir populations and then (ii) trigger efficient HIV reactivation, a prerequisite for any HIV-1 eradication strategy and the ultimate goal of this application.

抽象的 潜在的HIV-1感染已被认为是治愈HIV-1的发展的主要障碍 治疗，但是仍定义了控制潜在感染的宿主细胞病毒相互作用。此应用程序的关键是 意识到潜在的HIV-1感染事件的宿主细胞实际上以某种方式表型改变了 （i）迫使病毒进入潜在状态，并且（ii）使宿主细胞对刺激无反应， 从而防止HIV-1重新激活的有效治疗性诱导。这些变化超出了简单的 静态状态是功能记忆T细胞的特征。该建议将延长这些发现 并寻求解决艾滋病毒潜伏期研究领域的三个主要障碍。路障＃1（目标1） 涉及我们无法识别专门定义T细胞亚种群的生物标志物 感染的T细胞高度富集。这样的生物标志物将使我们能够详细介绍宿主的分子生物学 启用和维持潜在HIV-1感染的细胞状态。导致该应用程序，我们发现 细胞内变化允许感染潜在的HIV-1持续存在与唯一的CD4+CD28 –有关 CD9+ CD151+表型，一种T细胞表型，它也将小的CD4 T细胞亚种群划分为 HIV患者的ART患者增加。我们已经证明了CD151的表达本身与 水库容量。现在，我们将详细介绍CD28所描述的T细胞亚群的病毒储存能力 CD151和CD9表达。此应用程序中解决的障碍＃2（AIM 2）是一个问题 在潜在艾滋病毒感染事件的宿主细胞中观察到的细胞内变化（i）控制HIV转录和 （ii）抑制T细胞反应性。除了对宿主细胞因子/网络的拟议研究 控制潜在的艾滋病毒感染，我们已经确定了控制的几个目标/交互网络 潜在感染，这将立即调查。我们将进一步解决3号路障（AIM 3），问题 为什么来自HIV患者的刺激的离体T细胞材料似乎仅触发小型HIV-1重新激活 潜在的HIV感染细胞的一部分。根据我们的结果，我们预测T细胞亚群描述了 通过差异CD28，CD9和CD151表达模式，将表现出不同水平的无反应性 刺激，使潜在的艾滋病毒感染能够持续使用不同的有效性。根据结果​​我们有 获得了该应用程序以及我们将产生的其他结果，我们将立即开始 基于合理设计的基于复合的干预策略（i）首次重构T细胞响应能力 在水库种群中，然后（ii）触发有效的艾滋病毒重新激活，这是任何HIV-1消除的先决条件 策略和此应用程序的最终目标。