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: 9980780
• 负责人: OLAF KUTSCH
• 金额: $ 64.79万
• 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 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9980780
• 关键词: AIDS therapy; ATAC-seq; Address; Affect; BACH2 gene; Binding; Biological Markers; CD28 gene; CD4 Positive T Lymphocytes; CRISPR/Cas technology; 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 cell differentiation; 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 HIV reservoir; 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.

摘要 潜伏的HIV-1感染已被认为是开发治愈的HIV-1的主要障碍 治疗，但控制潜伏感染的宿主细胞-病毒相互作用仍然定义不清。此应用程序的关键是 意识到潜伏的HIV-1感染事件的宿主细胞实际上在某种程度上发生了表型改变 (I)迫使病毒进入潜伏状态，以及(Ii)使宿主细胞对刺激无反应， 从而防止有效的治疗性诱导HIV-1重新激活。这些变化超越了简单的 功能记忆T细胞所特有的一种静止状态。这项提案将对这些调查结果进行扩展 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) 担心我们无法识别特定定义T细胞亚群的生物标记物 感染的T细胞高度浓缩。这样的生物标记物将使我们能够详细了解宿主的分子生物学 启用和维持潜在HIV-1感染的细胞状态。在此应用程序中，我们发现 允许潜伏的HIV-1感染持续存在的细胞内变化与独特的CD4+CD28- CD9+CD151+表型，这是一种T细胞表型，也划分了一个小的CD4T细胞亚群 接受抗逆转录病毒治疗的HIV患者增加。我们已经证明CD151的表达本身与 水库容量。我们现在将详细描述CD28所描述的T细胞亚群的病毒储存能力， CD151和CD9的表达。本应用程序中解决的障碍#2(目标2)是如何 在潜伏的HIV感染事件的宿主细胞中观察到的细胞内变化作用(I)控制HIV转录和 和(Ii)抑制T细胞的反应性。除了对宿主细胞因子/网络的拟议研究之外 控制潜在的艾滋病毒感染，我们已经确定了几个目标/互动网络来控制 潜伏感染，并将立即进行调查。我们将进一步解决路障#3(目标3)，即 为什么体外刺激HIV患者的T细胞材料似乎只在很小的程度上触发HIV-1的重新激活 潜伏感染艾滋病毒的细胞的一部分。根据我们的结果，我们预测T细胞亚群描述 通过不同的CD28、CD9和CD151表达模式，将表现出不同程度的对 刺激，使潜伏的艾滋病毒感染以不同的效果持续存在。根据我们已有的结果 所获得的导致这一申请和我们将产生的额外结果，我们将立即开始 合理设计基于化合物的干预策略，首先重建T细胞的反应性 然后(Ii)触发有效的艾滋病毒重新激活，这是根除艾滋病毒-1的先决条件 策略和此应用程序的最终目标。