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 潜伏性HIV-1感染已被认为是开发治愈性HIV-1的主要障碍 治疗，但宿主细胞-病毒的相互作用，控制潜伏感染仍然不明确。此应用程序的关键是 认识到潜伏的HIV-1感染事件的宿主细胞实际上以一种 （i）迫使病毒进入潜伏状态和（ii）使宿主细胞对刺激无反应， 从而阻止HIV-1再活化的有效治疗诱导。这些变化超越了简单的 静止状态是功能性记忆T细胞的特征。这项建议将根据这些调查结果加以扩展 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） 我们无法鉴定特异性定义T细胞亚群的生物标志物， 感染的T细胞高度富集。这些生物标记物将使我们能够详细了解宿主的分子生物学 细胞状态，使和维持潜伏的HIV-1感染。导致这个应用程序，我们发现， 细胞内的变化使潜伏的HIV-1感染持续存在，这与独特的CD4 + CD28- CD9 + CD151+表型，一种T细胞表型，其也划分了一个小的CD4 T细胞亚群， 我们已经证明，CD151表达本身与抗逆转录病毒治疗相关， 水库容量我们现在将详细描述由CD28描述的T细胞亚群的病毒储库能力， CD151和CD9表达。在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（i）控制HIV转录， 和（ii）抑制T细胞应答性。除了对宿主细胞因子/网络的拟议研究外， 控制潜伏的艾滋病毒感染，我们已经确定了几个目标/相互作用网络，控制 潜伏感染，并将立即进行探测。我们将进一步解决障碍#3（目标3）， 为什么刺激HIV患者的离体T细胞物质似乎只会在一个小的细胞内触发HIV-1的重新激活， 潜伏性HIV感染细胞的一部分。基于我们的研究结果，我们预测T细胞亚群描述 不同的CD28，CD9和CD151表达模式，将表现出不同水平的无反应性， 刺激，使潜伏的艾滋病毒感染持续不同的功效。根据我们得到的结果 我们将立即开始， 合理设计基于化合物的干预策略，（i）首先重建T细胞反应性 然后（ii）触发有效的HIV重新激活，这是任何HIV-1根除的先决条件 战略和最终目标的应用。