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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要潜伏病毒库是治愈艾滋病毒的主要障碍，但人们对这种病毒库的性质知之甚少。调节它的分子机制。越来越多的证据表明，该水库的规模和性质受到滥用药物的影响。大麻素 (CB) 滥用在艾滋病毒感染者中尤其普遍（PWH），但 CB 对潜在 HIV 储存库的影响尚未得到研究。众所周知，CB 有通过激活广泛应用的 CB2 受体发挥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 依赖性改变潜伏 HIV 病毒库的大小、位置和转录组表型 CD4 T 细胞中 AP-1 转录因子的激活。与这一假设相一致的是，我们最近发现大麻素可促进 T 细胞模型中潜伏期的 HIV 重新激活。定义影响潜在 HIV 储存库上的 CB 对于设计适当的方法来清除储存库至关重要。使用 CB 的 PWH。为了实现这一目标，我们建议使用单细胞领域的尖端方法多组学分析来表征 CB 对潜在 HIV 储存库的影响，并测试其功能 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潜伏期模型确定CB暴露对潜伏感染细胞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 检测来量化一个完整的 HIV 储存库的大小和位置。使用 CB 的 PWH 队列与不使用 CB 的 PWH 进行比较。对于 R33 阶段，我们将结合以下结果这些研究通过详细的单细胞基因组分析来识别 PBMC 中 CB 调节的转录物，来自使用 CB 的 PWH 队列的 CD4 T 细胞。通过整合这些数据集，我们的目标是识别 CB 监管的调节 HIV 储存库大小和亚细胞位置的途径。总体而言，这些结果将向前推进我们对CB如何在分子水平上与潜在的HIV病毒库相互作用的理解。