Mechanisms of HIV persistence in distinct memory CD4+ T cell subsets in blood and

HIV在血液和血液中不同记忆CD4 T细胞亚群中持续存在的机制

• 批准号: 8880560
• 负责人: Nicolas Chomont
• 金额: $ 47.25万
• 依托单位: VGTI FLORIDA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-15 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8880560
• 关键词: Anti-Retroviral Agents; Autologous; Biological Assay; Blood; CD4 Positive T Lymphocytes; Cell Line; Cells; Clinical Trials; Coculture Techniques; Colon; Conflict (Psychology); DNA Methylation; DNA Modification Process; Data; Development; Distal part of ileum; Drug Targeting; Ensure; Epigenetic Process; Exhibits; Florida; Frequencies; Gene Expression; Genes; Genetic Transcription; Goals; HIV; HIV Genome; HIV Infections; Heterogeneity; Histone Acetylation; Histone Deacetylation; In Vitro; Individual; Lead; Life; Linear Regressions; Maintenance; Measures; Memory; Methylation; Molecular; Pathway interactions; Pharmaceutical Preparations; Population; Production; RNA Sequences; Rectum; Regression Analysis; Research Project Grants; Source; T-Lymphocyte Subsets; Testing; Tissues; Transcript; Vial device; Viral; Viral Genes; Viral Genome; Viremia; Virus; Virus Latency; antiretroviral therapy; base; cellular targeting; design; killings; lymph nodes; memory CD4 T lymphocyte; novel; novel therapeutics; public health relevance; research study; terminally differentiated effector memory (TEM) T cells; transcription factor; transcriptomics; viral RNA

DESCRIPTION (provided by applicant): It is now clear that antiretroviral therapy (ART) alone does not eradicate HIV: Even after more than 15 years of intensive and continuous therapy, the spread of the virus resumes within a few weeks upon cessation of ART in all but exceptional cases. A small pool of latently infected cells, usually referred as the "reservoir" of HIV infectio, provides a long-lived source of rebound viremia. Eradication of HIV has been hampered by the lack of information regarding the specific subsets harboring the viral reservoir and the mechanisms associated with the maintenance of latency. We have previously demonstrated that HIV persists in three memory T cell subsets displaying distinct functional and survival capacities, namely central (TCM) transitional (TTM) and effector (TEM) memory CD4+ T cells in the blood of virally suppressed subjects. Interestingly, our preliminary data indicate that these cell subsets also ensure HIV persistence in tissue reservoirs such as the lymph nodes, rectum, colon and terminal ileum. Therefore, there is an urgent need to better characterize these three cellular HIV reservoirs to achieve their eradication. The objective of this project is to identify mechanisms of action that can lead to the reactivation and subsequent elimination of latently infected cells in virally suppressed subjects. We hypothesize that the mechanisms responsible for HIV latency in TCM, TTM and TEM are different, and that these reservoirs should be targeted by different classes of anti-latency agents. We will first identify the molecular mechanisms responsible for HIV latency in TCM, TTM and TEM cells (Specific Aim 1). We will focus on epigenetic modification of the DNA through methylation and histone deacetylation, limited availability of critical transcription factors and inefficient elongation of the nascent vial transcripts. We will also use RNA sequencing to identify novel cellular pathways that correlate with the magnitude of the latent reservoir in the CD4+ T cell subsets from 24 virally suppressed subjects. We will then select the most efficient combination of compounds that will interfere with these mechanisms and allow elimination of all cellular reservoirs (Specific Aim 2). We have selected molecules that have shown promising effects in previous studies and/or that are currently tested in clinical trials. As it is highly likely that targeting a single cellular reservir or a single mechanism involved in viral latency will not be successful, we will test the ability of combinations of lead compounds from each class to disrupt latency in latently infected CD4+ T cells from virally suppressed subjects. Using a novel co-culture assay recently developed at VGTI Florida, we will assess if reactivation with this combination of anti-latency drugs can lead to the elimination of reservoir cells by autologous HIV specific CTL. The deliverable of this research project is a combination of 3-5 clinically tolerable agents that can reverse latency in authentic latently infected cells from virally suppressed subjects. The results from these studies will constitute the basis for the design of a clinical trial aimed at eradicating HIV.

描述（由申请人提供）：现在很明显，单靠抗逆转录病毒疗法（ART）并不能根除艾滋病毒：即使经过15年以上的强化和持续治疗，除了例外情况外，在停止ART后的几周内，病毒的传播仍会恢复。一小群潜伏感染的细胞，通常被称为HIV感染的“储存库”，是反弹性病毒血症的长期来源。艾滋病毒的根除受到阻碍，因为缺乏关于携带病毒库的特定亚群和与维持潜伏期有关的机制的信息。我们以前已经证明，HIV持续存在于三个记忆T细胞亚群中，表现出不同的功能和生存能力，即病毒抑制受试者血液中的中央（TCM）过渡（TTM）和效应（TEM）记忆CD 4 + T细胞。有趣的是，我们的初步数据表明，这些细胞亚群也确保了HIV在组织储库（如淋巴结、直肠、结肠和末端回肠）中的持久性。因此，迫切需要更好地描述这三种细胞HIV宿主，以实现其根除。该项目的目的是确定可以导致病毒抑制受试者中潜伏感染细胞的再激活和随后消除的作用机制。我们假设，负责艾滋病毒潜伏在中医，TTM和TEM的机制是不同的，这些水库应针对不同类别的抗潜伏剂。我们将首先确定负责TCM，TTM和TEM细胞中HIV潜伏期的分子机制（具体目标1）。我们将重点关注通过甲基化和组蛋白去乙酰化对DNA进行的表观遗传修饰，关键转录因子的有限可用性和新生小瓶转录物的低效延伸。我们还将使用RNA测序来鉴定与来自24名病毒抑制受试者的CD 4 + T细胞亚群中潜伏库的大小相关的新细胞途径。然后，我们将选择最有效的化合物组合，这些化合物将干扰这些机制，并允许消除所有细胞储库（具体目标2）。我们选择了在以前的研究中显示出有希望的效果和/或目前在临床试验中测试的分子。由于靶向单个细胞reservir或涉及病毒潜伏期的单个机制很可能不会成功，我们将测试每类先导化合物的组合破坏来自病毒抑制受试者的潜伏感染的CD 4 + T细胞中的潜伏期的能力。使用最近在VGTI佛罗里达开发的一种新的共培养试验，我们将评估这种抗潜伏药物组合的再激活是否可以通过自体HIV特异性CTL消除储库细胞。该研究项目的可交付成果是3-5种临床耐受药物的组合，这些药物可以逆转来自病毒抑制受试者的真实潜伏感染细胞的潜伏期。这些研究的结果将构成旨在根除艾滋病毒的临床试验设计的基础。