Defining the impact of cannabinoids on the latent HIV reservoir through multi-omic analysis

通过多组学分析确定大麻素对潜在 HIV 储存库的影响

• 批准号: 10219556
• 负责人: Edward P Browne
• 金额: $ 67.15万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10219556
• 关键词: Anti-Inflammatory Agents; Biological Assay; Biological Models; Blood specimen; CD4 Positive T Lymphocytes; CNR2 gene; Cannabinoids; Cell model; Cells; Cellular Assay; Characteristics; Chromatin Structure; Chronic; Clinical; Collection; DNA; Data; Data Set; Gene Expression; Gene Expression Profile; Genetic Transcription; Genomics; Goals; HIV; HIV Infections; Immune; In Vitro; Individual; Investigation; Location; Maintenance; Methods; Modeling; Molecular; Nature; Outcome; Pathway interactions; Peripheral Blood Mononuclear Cell; Phase; Phenotype; Population; Property; Proviruses; Role; Sampling; Shapes; Signal Pathway; Signal Transduction; T-Lymphocyte; Testing; Time; Transcript; Transcription Factor AP-1; Viral; Viral Genes; Viral reservoir; cohort; design; drug of abuse; epigenome; epigenomics; immunoregulation; in vivo; insight; latent HIV reservoir; multiple omics; non-cannabinoid; novel; reactivation from latency; tool; transcriptome; transcriptomics

Project summary The latent reservoir is the primary barrier to curing HIV, but little is known about the nature of this reservoir or the molecular mechanisms that regulate it. Increasing evidence suggests that the size and nature of this reservoir is impacted by drugs of abuse. Cannabinoid (CB) abuse, in particular, is prevalent amongst people with HIV (PWH), but the impact of CBs on the latent HIV reservoir has not been investigated. CBs are known to have immuno-modulatory and anti-inflammatory activities through activation of the CB2 receptor that is widely expressed in immune cells, including CD4 T cells. Our hypothesis is that CB exposure during HIV infection alters the size, location and transcriptomic phenotype of the latent HIV reservoir through CB2-dependent activation of the AP-1 transcription factor in CD4 T cells. Consistent with this hypothesis we have recently discovered that cannabinoids promote reactivation of HIV from latency in a T cell model. Defining the impact of CBs on the latent HIV reservoir will be critical to designing appropriate approaches to clear the reservoir from PWH who use CBs. To achieve this goal we propose to use cutting edge methods from the fields of single cell multi-omic analysis to characterize the effect of CBs on the latent HIV reservoir, and to test the functional role of CB-activated pathways in HIV latency. In the R61 phase, we will use a primary cell HIV latency model to define the impact of CB exposure on the transcriptome and epigenome of latently infected cells. Additionally, we will use a cutting-edge single cell HIV assay to quantify the size and location of the intact HIV reservoir in a cohort of CB-using PWH compared to non CB-using PWH. For the R33 phase, we will combine the results from these studies with a detailed single cell genomic analysis of identify CB-regulated transcripts in the PBMCs and CD4 T cells from the CB-using PWH cohort. By integrating these datasets we aim to identify CB-regulated pathways that regulate the size and subcellular location of the HIV reservoir. Overall these results will advance our understanding of how CBs interact with the latent HIV reservoir at the molecular level.

项目摘要 潜在储层是治愈艾滋病毒的主要障碍，但对此水库的性质知之甚少 调节它的分子机制。越来越多的证据表明该储层的大小和性质 受虐待药物的影响。尤其是大麻素（CB）滥用，在艾滋病毒患者中普遍存在 （PWH），但尚未研究CBS对潜在艾滋病毒储量的影响。哥伦比亚广播公司有 通过广泛的CB2受体激活的免疫调节和抗炎活性 在免疫细胞中表达，包括CD4 T细胞。我们的假设是HIV感染期间的CB暴露 通过CB2依赖性改变潜在的HIV库的大小，位置和转录表表型 CD4 T细胞中AP-1转录因子的激活。与这个假设一致，我们最近有 发现大麻素促进了T细胞模型中潜伏期的HIV重新激活。定义 潜在艾滋病毒水库上的哥伦比亚广播公司对于设计适当的方法以清除水库从 使用CBS的PWH。为了实现此目标，我们建议使用单个单元格的尖端方法 多矩分析以表征CBS对潜在艾滋病毒库的影响，并测试功能 CB激活途径在HIV潜伏期中的作用。在R61阶段，我们将使用原代细胞HIV潜伏期模型 定义CB暴露对潜在感染细胞的转录组和表观基因组的影响。此外， 我们将使用尖端的单细胞HIV分析来量化完整的HIV储存库的大小和位置 与非CB使用PWH相比，CB使用PWH的队列。对于R33阶段，我们将结合 这些研究对PBMC中CB调节的转录本的详细单细胞基因组分析和 来自CB使用PWH队列的CD4 T细胞。通过集成这些数据集，我们旨在识别CB调节 调节HIV储层的大小和亚细胞位置的途径。总的来说这些结果将进步 我们对CBS如何在分子水平与潜在的HIV储藏室相互作用的理解。