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

项目概要（摘要） 在患有精神分裂症的人中，将大麻用于娱乐和医疗目的的比例高得不成比例。 艾滋病毒（PLWH）和近一半的大麻使用PLWH估计有大麻使用障碍的风险。然而， 大麻是否对PLWH的中枢神经系统（CNS）有治疗作用或有害， 有争议的，强调需要良好的对照研究，从特定的 大麻素、大脑区域和CNS细胞类型。我们的研究表明，细胞类型特异性表观遗传 模式与艾滋病毒相关的认知障碍PLWH，更频繁或最近的大麻使用可能 减少骨髓炎症和影响PLWH的脑结构，以及我们最近对CNS的单细胞研究 在PLWH中显示了不同的CSF小胶质细胞样细胞表达CD204，在PLWH和正在进行的HIV病毒感染中， 先前的研究表明，神经源性脑区 包括侧脑室和海马的脑室下区与持续性 艾滋病毒感染和大麻素暴露。然而，在理解神经原性大脑区域方面的关键差距， 单细胞水平的艾滋病毒和大麻素暴露的设置仍然存在。我们利用脑组织 在HIV的非人灵长类动物（NHP）模型中建立的大麻二酚和THC的口服给药模型， 新的单细胞技术的应用，允许同时分析基因表达和开放 脑组织中来自同一细胞的染色质（10X Genomics Multiome：RNA + ATAC），一种新的单细胞测定法 能够测量多种组蛋白修饰和当前ART方案的药理学特征， 大脑组织中的大麻素水平。我们的中心假设是大麻素在改善 通过增强神经祖细胞的增殖和存活， 不成熟的神经元和减少神经胶质炎症。识别细胞类型、表观遗传细胞状态和基因 与神经发病机制、病毒持续存在和大麻素暴露相关的途径，我们正在利用180 来自大麻二酚（CBD）或Δ 9- 四氢大麻酚（THC）在NHP和单细胞测定（10X Genomics单核多组和新的 在NYGC开发的单细胞试验能够测量多个组蛋白的全基因组存在 修饰和蛋白质-DNA结合位点）。此外，伴随的单细胞数据将从 基于HIV状态的来自40个供体的人类死后脑组织的保守神经原性脑区域 （+/-）和大麻暴露（+/-）。我们还将探讨在神经源性区域的单细胞的频率， 通过利用生物信息学管道来检测大麻素对大脑的感染和影响， 病毒转录物和转座酶可接近的前病毒。该项目将产生全面的单细胞 NHP和人类的数据集，以提高我们对HIV和大麻素之间的相互作用的理解， 大脑的神经原性区域，并对SCORCH计划扩展的目标具有高度的计划优先级。

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.