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

项目摘要 潜伏的水库是治愈艾滋病毒的主要障碍，但很少有人知道这个水库的性质， 调节它的分子机制。越来越多的证据表明，这个水库的大小和性质， 受到药物滥用的影响。特别是大麻素（CB）滥用在艾滋病毒感染者中普遍存在 (PWH)但CBs对潜伏HIV库的影响尚未研究。据了解，CB具有 免疫调节和抗炎活性，通过激活CB 2受体，广泛 在免疫细胞中表达，包括CD 4 T细胞。我们的假设是，在艾滋病毒感染期间接触CB 通过CB 2依赖性改变潜伏HIV储库的大小、位置和转录组表型， 活化CD 4 T细胞中的AP-1转录因子。与这个假设相一致，我们最近 发现大麻素在T细胞模型中促进HIV从潜伏期重新激活。定义影响 对潜伏的艾滋病毒库的CB将是至关重要的，以设计适当的方法来清除水库， 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，以表征CB对潜伏HIV储库的影响，并测试CB的功能性。 CB激活通路在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的转录组和表观基因组的影响。此外，本发明还 我们将使用最先进的单细胞HIV检测来量化完整HIV库的大小和位置， 使用CB的PWH队列与不使用CB的PWH队列的比较。对于R33阶段，我们将联合收割机的结果与 这些研究通过详细的单细胞基因组分析来鉴定PBMC中CB调节的转录物， 来自使用CB的PWH队列的CD 4 T细胞。通过整合这些数据集，我们的目标是确定CB调节的 调节HIV储库大小和亚细胞位置的途径。总的来说，这些结果将促进 我们对CB如何在分子水平上与潜伏的HIV库相互作用的理解。

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.