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

总结 阿片类药物的滥用与治疗不依从性，病毒传播的风险更大， 艾滋病的临床进展更快。本项目背后的首要假设是 在药物滥用相关的单细胞水平上分析神经元和神经胶质细胞的分子谱， 通过单细胞核RNA-Seq（snRNA-Seq）对大脑区域的研究，将揭示出由细胞核调控失调的关键基因。 艾滋病毒与阿片类药物滥用的相互作用，导致神经变性和认知障碍。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进行重建， 结合snRNA-Seq从HIV中探询基因组规模整合基因调控网络 转基因（Tg）大鼠，其携带表达慢性低水平的多种HIV-1转基因， 疾病相关细胞类型和野生型大鼠中的HIV-1蛋白。 偶尔但有限使用药物在临床上与依赖性药物使用不同，依赖性药物使用是 其特点是出现依赖性和消极的情绪状态时，获得药物是 这会驱动负强化，这是寻求毒品的强大动力来源。所以我们 将使用最先进的自愿静脉注射阿片类药物自我管理模式， (ShA)条件，其特点是非依赖性的，“娱乐性”的药物使用模式， 获得（LGA）条件，导致依赖性药物摄入。LgA条件下的药物摄入量增加 它与人类物质使用障碍（SUD）高度相关，因为它模拟了所有7种 精神障碍诊断和统计手册（DSM）-IV和11个中的7个中的药物成瘾标准 我们发现HIV Tg大鼠在这种LgA范例中自我施用羟考酮， 增加的自我给药显示出增加的神经损伤和认知损害。 该项目将解决以下关于艾滋病毒感染背景下阿片类药物滥用的棘手问题： 驱动神经炎症、神经变性、病毒表达和 在HIV背景下，阿片类药物自我给药和认知障碍的升级？ 总的来说，这个跨学科的合作提案整合了单细胞水平的转录组学，国家的- 在HIV Tg和野生型大鼠中的艺术行为方法，以及用于对 单细胞水平的基因调控网络将阐明HIV作用的关键机制 和阿片类药物滥用及其对神经HIV进展、病毒表达和 持久性。结果将表明变革性的新机制假设，可能会导致新的 在艾滋病毒背景下阿片类药物使用障碍（OUD）的治疗概念，并将为 neuroHIV字段将通过SCORCH数据协调中心和其他公共 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.