Elucidating single cell changes in neurogenic brain regions during HIV and cannabinoid exposure

项目摘要(摘要) 将大麻用于娱乐和医疗目的在吸毒者中高得不成比例 据估计，艾滋病毒(PLWH)和近一半使用PLWH的大麻有患大麻使用障碍的风险。然而， 大麻对PLWH的中枢神经系统(CNS)是治疗还是有害仍然存在 有争议的，强调了从特定的数据中产生可重现数据的良好控制研究的必要性 大麻素、脑区和中枢神经系统细胞类型。我们的研究表明，细胞类型特异的表观遗传学 与PLWH中艾滋病毒相关的认知障碍有关的模式，更频繁或最近使用大麻可能 减少PLWH的髓系炎症和影响脑结构，以及我们最近对中枢神经系统的单细胞研究 在PLWH中发现在PLWH和正在进行的HIV病毒中有明显的表达CD204的脑脊液小胶质细胞样细胞 脑脊液细胞中的转录，尽管有抗逆转录病毒治疗。先前的研究表明，神经源性脑区 包括侧脑室和海马室下区与持续性密切相关 艾滋病毒感染和大麻素暴露。然而，在理解神经源性脑区方面存在关键差距 艾滋病毒和大麻素暴露环境中的单细胞水平仍然存在。我们正在利用来自 在非人类灵长类(NHP)HIV模型中建立了大麻二醇和THC的口服剂量模型， 允许同时分析基因表达和开放的新的单细胞技术的应用 脑组织中来自同一细胞的染色质(10X基因组多组体：RNA+ATAC)，一种新的单细胞分析方法 能够测量多种组蛋白修饰，以及当前ART方案和 脑组织中的大麻素水平。我们的中心假设是大麻素的治疗作用 HIV通过促进神经前体细胞的增殖和存活在中枢神经系统中的神经发病机制 幼稚的神经元和减少神经胶质炎症。鉴定细胞类型、表观遗传细胞状态和基因 与神经致病、病毒持久性和大麻素暴露相关的途径，我们正在利用180 口服大麻二醇或Δ9的脑组织样本- 四氢大麻酚(THC)在NHP和单细胞分析中的应用(10X基因组学、单核多核体和一种新的 NYGC开发的单细胞分析方法能够测量全基因组中多个组蛋白的存在 修饰和蛋白质-DNA结合位点)。此外，附带的单元格数据将从 基于HIV状况的40例死后人类脑组织中保守的神经源性脑区 (+/-)和大麻暴露(+/-)。我们还将探索单个细胞在脑内神经源性区域的频率。 通过利用生物信息学管道检测受大麻类药物感染和影响的大脑 病毒转录本和转座酶可访问的前病毒。该项目将产生全面的单细胞 NHP和人类中的数据集，以提高我们对艾滋病毒和大麻类物质之间的相互作用的理解 脑部的神经源性区域，并对SCOCH计划扩展的目标具有较高的计划优先级。

PROJECT SUMMARY (ABSTRACT) The use of cannabis for recreation and medicinal purposes is disproportionately high among people living with HIV (PLWH) and nearly half of cannabis using PLWH are estimated to be at risk for cannabis use disorder. Yet, whether cannabis is therapeutic or detrimental on the central nervous system (CNS) of PLWH remains controversial, highlighting the need for well-controlled studies generating reproducible data from specific cannabinoids, brain regions, and CNS cell types. Our research has shown that cell-type specific epigenetic patterns relate to HIV-associated cognitive impairment in PLWH, more frequent or recent cannabis use may reduce myeloid inflammation and impact brain structure in PLWH, and our recent single cell studies of the CNS in PLWH revealed distinct CSF microglia-like cells expressing CD204 in PLWH and ongoing HIV viral transcription in cerebrospinal fluid cells despite ART. Prior research has shown neurogenic brain regions including the subventricular zone of the lateral ventricles and hippocampus are of high relevance to persistent HIV infection and cannabinoid exposures. However, critical gaps in understanding neurogenic brain regions at single cell level in the setting of HIV and cannabinoid exposures remain. We are leveraging brain tissues from an established oral dosing model of cannabidiol and THC in a nonhuman primate (NHP) model of HIV, application of new single cell technologies permitting the simultaneous profiling of gene expression and open chromatin from the same cell (10X Genomics Multiome: RNA+ATAC) in brain tissues, a new single cell assay capable of measuring multiple histone modifications, and pharmacological profiling of current ART regimens and cannabinoid levels in brain tissues. Our central hypothesis is a therapeutic role of cannabinoids in ameliorating HIV neuropathogenesis in the CNS by enhancing the proliferation and survival of neural progenitor cells and immature neurons and reducing glial inflammation. To identify cell types, epigenetic cell states, and gene pathways relevant to neuropathogenesis, viral persistence, and cannabinoid exposures, we are harnessing 180 brain tissue samples from an established oral administration of either cannabidiol (CBD) or Δ9- tetrahydrocannabinol (THC) in NHP and single cell assays (10X Genomics single nucleus multiome and a new single cell assay developed at the NYGC capable of measuring the genome-wide presence of multiple histone modifications and protein-DNA binding sites). Moreover, accompanying single cell data will be generated from conserved neurogenic brain regions of human postmortem brain tissues from 40 donors based on HIV status (+/-) and cannabis exposure (+/-). We will also explore the frequency of single cells in neurogenic regions of the brain that are infected and impacted by cannabinoids by harnessing a bioinformatics pipeline that detects both viral transcripts and transposase accessible provirus. This project will generate comprehensive single cell datasets in NHP and humans to improve our understanding of the cross talk between HIV and cannabinoids in neurogenic regions of the brain and has high programmatic priority to goals of the SCORCH program expansion.