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

项目总结 潜伏的蓄水池是治愈艾滋病毒的主要障碍，但人们对这种蓄水池的性质或 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 受到滥用药物的影响。特别是在艾滋病毒携带者中，滥用大麻素(CB)很普遍。 (PWH)，但CBS对潜伏的艾滋病毒宿主的影响尚未调查。众所周知，哥伦比亚广播公司拥有 通过激活广泛存在的CB2受体发挥免疫调节和抗炎作用 在免疫细胞中表达，包括CD4T细胞。我们的假设是，在HIV感染期间接触CB 通过依赖CB2改变潜伏的HIV储备库的大小、位置和转录表型 CD4T细胞中AP-1转录因子的激活。与我们最近得到的这一假设一致 发现在T细胞模型中，大麻类物质促进艾滋病毒从潜伏期重新激活。定义以下方面的影响 CBS关于潜伏的艾滋病毒宿主的研究将对设计适当的方法来清除该宿主至关重要 使用CBS的PWH。为了实现这一目标，我们建议使用来自单细胞领域的尖端方法 多组学分析表征CBS对HIV潜伏宿主的作用，并检验其功能 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 明确CB暴露对潜伏感染细胞的转录组和表观基因组的影响。另外， 我们将使用尖端的单细胞艾滋病毒检测来量化完整的艾滋病毒储存库的大小和位置 CB-使用PWH与非CB-使用PWH的队列比较。对于R33阶段，我们将结合以下结果 这些研究用详细的单细胞基因组分析确定了PBMCs和PBMC中CB调节的转录本 CD4T细胞来自CB-使用PWH队列。通过整合这些数据集，我们的目标是识别受CB监管的 调节艾滋病毒储存库大小和亚细胞位置的途径。总体而言，这些结果将取得进展 我们对CBS如何在分子水平上与潜在的艾滋病毒宿主相互作用的理解。

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.