Single Cell Transcriptomics of the Cocaine Use Disorder in the Context of HIV

项目总结 HIV感染者的系统免疫激活被认为是导致较高发病率的原因 慢性炎症性疾病，包括艾滋病毒相关的神经认知障碍(手)。急性HIV感染 在中枢神经系统中，被认为启动了一连串的促炎事件，导致炎症诱导 神经元损伤和相关的神经认知障碍，即使在目前的组合中也是明显的 抗逆转录病毒治疗(CART)时代。使用精神刺激剂(如可卡因和甲基苯丙胺)和 研究表明，酒精会破坏血脑屏障的完整性。血脑屏障受损可能会增加免疫细胞渗透到 并促进神经胶质细胞的激活，增加炎症和神经毒性。有趣的是，渗透性增加 血脑屏障的异常与HIV神经功能障碍的进展有关。因此，联合作用的结果是 可卡因的使用和艾滋病毒感染可能会对血脑屏障的破坏产生附加作用，并进一步影响与艾滋病毒相关的 神经认知障碍。然而，在全基因组水平上进行的分子水平的研究还很少。 了解BBB在物质使用障碍和HIV感染/手中的完整性。拟议的研究将 解决这个重要的问题。我们的中心假设是，滥用可卡因会加剧艾滋病毒的致病作用 在中枢神经系统中，通过破坏血脑屏障和失调大脑中的神经胶质细胞数量。我们的整体 目的是利用患者脑中单个核水平的细胞类型特异性转录信息。 表征可卡因使用障碍对中枢神经系统神经元和神经胶质细胞、HIV感染和 举起手来。我们将确定单核基因表达的特征，并识别失控的基因调控网络 在每个与艾滋病毒感染者和/或滥用可卡因有关的神经元和神经胶质细胞群体中 用手。我们还将进行计算分析，以确定神经元和神经胶质细胞的调节网络 因滥用可卡因而改变。在验证和功能表征组件中，我们将描述TOP 基因在3D脑器官模型中，将以CRISPR基因敲除和过表达为特征。 这些目标的成功完成将产生重大的研究和临床影响，1)阐明 可卡因滥用改变了中枢神经系统中HIV/Hand的发病机制，2)发现候选分子进行调节 艾滋病毒感染或持续在中枢神经系统滥用可卡因的情况。

PROJECT SUMMARY Systemic immune activation in people living with HIV has been hypothesized to account for higher incidence of chronic inflammatory diseases, including HIV-associated neurocognitive disorders (HAND). Acute HIV infection in the CNS is thought to initiate a cascade of pro-inflammatory events that result in inflammation-induced neuronal injury and associated neurocognitive disorders that are evident even in the present combination antiretroviral therapy (cART) era. The use of psychostimulants (such as cocaine and methamphetamine) and alcohol has been shown to disrupt BBB integrity. Disrupted BBB may increase immune cell infiltrating into the CNS and promote glial activation, increased inflammation and neurotoxicity. Interestingly, increased permeability of BBB has been implicated in the progression of HIV neurological dysfunction. Thus, the combined effect of cocaine usage and HIV infection can cause an additive effect on BBB disruption and further impact HIV-related neurocognitive impairments. However, not much genome-wide molecular level study has been done in understanding BBB integrity in substance use disorder and in HIV infection/HAND. The proposed study will address this important question. Our central hypothesis is that cocaine misuse exacerbates HIV pathogenesis in the CNS by disrupting blood-brain barrier and dysregulating the glial population in the brain. Our overall objective is to exploit cell type specific transcriptomic information at the single nuclei level from patient brain samples to characterize the effects of cocaine use disorder on CNS neuronal and glial cells, HIV infection and HANDs. We will characterize single nuclei gene expression and identify dysregulated gene regulatory networks in each of the neuronal and glial populations associated with cocaine misuse in HIV infected individuals and/or with HANDs. We will also perform computational analysis to identify neuronal and glial cell regulatory networks altered by cocaine misuse. In the validation and functional characterization component, we will characterize top genes in 3D brain organoid model and will characterize with CRISPR knockout and overexpression of the gene. Successful completion of these aims will have significant research and clinical impact by 1) elucidating how cocaine misuse alters HIV/HAND pathogenesis in the CNS, and 2) discovering candidate molecules to regulate HIV infection or persistence in the CNS in the context of cocaine misuse.