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 潜伏性 HIV-1 感染已被认为是开发治疗性 HIV-1 药物的主要障碍 治疗，但控制潜伏感染的宿主细胞-病毒相互作用仍然不明确。这个应用程序的关键是 认识到潜伏 HIV-1 感染事件的宿主细胞实际上以某种方式发生表型改变 (i) 迫使病毒进入潜伏状态，并且 (ii) 使宿主细胞对刺激无反应， 从而阻止有效治疗诱导 HIV-1 重新激活。这些变化不仅仅是一个简单的改变 静止状态是功能性记忆 T 细胞的特征。该提案将扩展这些发现 并寻求解决艾滋病毒潜伏期研究领域的三大障碍。路障 #1（目标 1） 涉及我们无法识别特异性定义 T 细胞亚群的生物标志物，其中潜在的 T 细胞亚群 受感染的 T 细胞高度富集。这些生物标志物将使我们能够详细了解宿主的分子生物学 能够实现并维持潜在 HIV-1 感染的细胞状态。在这个应用程序中，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，一种 T 细胞表型，也划分了一个小的 CD4 T 细胞亚群 接受 ART 治疗的 HIV 患者数量有所增加。我们已经证明 CD151 表达本身与 水库容量。我们现在将详细介绍 CD28 描述的 T 细胞亚群的病毒库容量， CD151 和 CD9 表达。本申请中解决的障碍#2（目标 2）是如何 在潜伏 HIV 感染事件的宿主细胞中观察到的细胞内变化起到 (i) 控制 HIV 转录和 (ii) 抑制 T 细胞反应性。除了拟议的宿主细胞因子/网络研究之外 控制潜在的艾滋病毒感染，我们已经确定了几个控制潜在艾滋病毒感染的目标/相互作用网络 潜在感染，将立即进行调查。我们将进一步解决障碍#3（目标 3），即问题 为什么刺激来自 HIV 患者的离体 T 细胞材料似乎只能在小范围内触发 HIV-1 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，我们预测描述的 T 细胞亚群 通过差异化 CD28、CD9 和 CD151 表达模式，将表现出不同程度的无反应性 刺激，使潜伏的艾滋病毒感染持续存在，并具有不同的功效。根据我们得到的结果 获得导致此申请以及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，该策略将 (i) 首先重建 T 细胞反应性 在储存人群中，然后 (ii) 触发有效的 HIV 重新激活，这是根除 HIV-1 的先决条件 该应用程序的策略和最终目标。