Single nucleus gene expression in moderate and compulsive opioid self-administration in a rodent model of HIV

摘要 阿片类药物的滥用与治疗不依从、更大的病毒传播风险、 并且HIV疾病的临床进展更快。本项目背后的首要假设是 与药物滥用相关的单细胞水平的神经细胞和神经胶质细胞的分子图谱分析 单核RNA-Seq(SnRNA-Seq)的大脑区域将揭示受 艾滋病毒与阿片类药物滥用的相互作用，导致神经退化和认知障碍。要测试 目前的假设，我们建议使用经过验证的系统生物学策略来重建和 结合人类免疫缺陷病毒SnRNA-Seq的基因组级整合基因调控网络的构建 转基因(TG)大鼠，携带非复制型HIV-1转基因表达慢性低水平的 疾病相关细胞类型中的HIV-1蛋白，以及野生型老鼠。 偶尔但有限的药物使用在临床上不同于依赖药物的使用，后者是 以出现依赖和当获得药物时的负面情绪状态为特征的 防止这种情况会导致负面强化，这是寻求毒品的强大动力来源。因此，我们 将在短时间内使用最先进的自愿静脉注射阿片类药物自我给药范例 (Sa)以非依赖、“娱乐”的吸毒模式为特征的条件，以及长期 进入(LGA)条件，导致依赖药物摄取。在LGA条件下增加的药物摄入量是 与人类物质使用障碍(SUD)高度相关，因为有人建议它模拟所有7个 《精神疾病诊断和统计手册》(DSM)中的药物成瘾标准--第四部分和第七部分 DSM-V中的标准我们发现HIV TG大鼠在这种LGA范式中自我给药羟考酮 自我管理升级会增加神经损伤和认知障碍。 该项目将解决关于在艾滋病毒感染环境中滥用阿片类药物的下列令人烦恼的问题： 导致神经炎症、神经退行性变、病毒表达和 艾滋病毒环境中阿片类药物自我管理和认知障碍的升级？ 总体而言，这一跨学科的协作性提案整合了单细胞水平的转录组学、最新进展- 在HIV TG和野生型大鼠中的ART行为方法以及去卷积的计算策略 单细胞水平的基因调控网络将阐明艾滋病毒影响的关键机制 和阿片类药物滥用及其对神经艾滋病毒进展、病毒表达和 坚持不懈。结果将表明变革性的新的机械论假说可能导致新的 艾滋病毒背景下阿片使用障碍(OUD)的治疗概念，并将为 将通过SCARCH数据协调中心和其他公共机构公开提供神经艾滋病毒领域 储存库。

Summary The abuse of opioid drugs is associated with treatment non-compliance, greater risk of viral transmission, and more rapid clinical progression of HIV disease. The overarching hypothesis behind the present project is that the analysis of molecular profiles of neuronal and glia cells at the single cell level in drug abuse-relevant brain regions by single nucleus RNA-Seq (snRNA-Seq) will reveal key genes that are dysregulated by the interaction of HIV with opioid abuse, resulting in neurodegeneration and cognitive impairment. To test the present hypothesis, we propose to use validated systems biology strategies for the reconstruction and interrogation of a genome-scale integrated gene regulatory network in conjunction with snRNA-Seq from HIV transgenic (Tg) rats, which harbor a non-replicating HIV-1 transgene expressing chronic low-levels of multiple HIV-1 proteins in disease-relevant cell types, and wild-type rats. The occasional but limited use of a drug is clinically distinct from dependent drug use, which is characterized by the emergence of dependence and a negative emotional state when access to the drug is prevented that drives negative reinforcement, a powerful source of motivation for drug seeking. Therefore, we will use a state-of-the-art paradigm of voluntary intravenous opioid self-administration under short access (ShA) conditions, which is characterized by a non-dependent, “recreational” pattern of drug use, and long access (LgA) conditions, which leads to dependent drug intake. Escalated drug intake under LgA conditions is highly relevant to human substance use disorder (SUD) as it has been suggested that it models all 7 of the criteria for drug addiction in the Diagnostic and Statistical Manual of Mental Disorders (DSM)-IV and 7 of the 11 criteria in the DSM-V. We showed that HIV Tg rats self-administering oxycodone in this LgA paradigm of escalated self-administration display increased neural injury and cognitive impairment. The project will address the following vexing question about opioid abuse in the setting of HIV infection: what are the cell types and cell states that drive neuroinflammation, neurodegeneration, virus expression, and escalated (dependent) opioid self-administration and cognitive impairment in the setting of HIV? Overall, this collaborative interdisciplinary proposal integrating single cell level transcriptomics, state-of-the- art behavior methods in HIV Tg and wild-type rats, and computational strategies for the deconvolution of the gene regulatory network at the single cell level will elucidate key mechanisms that underlie the effects of HIV and opioid abuse and their detrimental interactions on neuroHIV progression, virus expression and persistence. The results will indicate transformative new mechanistic hypotheses that may lead to novel therapeutic concepts for opioid use disorder (OUD) in the setting of HIV and will establish key resources for the neuroHIV field to be made publicly available through the SCORCH data coordination center and other public repositories.