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

总结 滥用甲基苯丙胺等兴奋剂加剧了艾滋病毒的有害影响 感染在这里，我们将进行单核RNA-Seq，目标是鉴定细胞类型和细胞状态 这是艾滋病毒和慢性甲基苯丙胺（METH）自我管理对关键大脑的影响的关键 与HIV转基因（Tg）大鼠中持续HIV感染和METH使用障碍的影响相关的区域， 其携带非复制型HIV-1转基因并表达慢性低水平的多种HIV-1蛋白。的 偶尔但有限地使用药物在临床上不同于逐渐增加的药物使用，其特征在于 出现慢性强迫性寻药和吸毒。因此，我们将使用一个既定的，最先进的 在长期使用（LgA）条件下自愿静脉内药物自我给药的范例，导致 与短期接触（ShA）条件下自我给药相比，逐步增加（强迫性）METH摄入量， 这导致适度和稳定的甲基苯丙胺摄入模式。药物摄入量增加的范例 LgA条件与人类物质使用障碍（SUD）高度相关，因为它模拟了所有7个标准 精神疾病诊断和统计手册（DSM）-IV和11项标准中的7项 我们发现，HIV Tg大鼠自我管理METH在这个范例显示增加 强迫症、神经炎症和神经损伤。该项目将解决以下令人烦恼的问题， CNS中持续的HIV感染：驱动神经炎症的细胞类型和细胞状态是什么， 神经变性，和强迫性METH滥用在艾滋病毒的设置，可以揭示致病 neuroHIV疾病进展、病毒表达和持久性背后的机制？总体 本项目背后的假设是，在单细胞基因调控网络的探索 水平将阐明艾滋病毒和甲基苯丙胺滥用及其有害影响的关键机制 神经HIV疾病进展，病毒表达和病毒持久性的相互作用，并将表明新的 神经炎症、神经变性和强迫性服用METH的治疗靶点。为了验证这一 假设，我们将使用一个有效的系统生物学策略，重建和询问 全基因组基因调控网络，以确定与影响相关的基因网络失调 艾滋病毒，强制性甲基苯丙胺的使用，以及它们在单细胞分辨率基因分析中的相互作用 RNA测序总的来说，这个跨学科的合作提案整合了单细胞水平的转录组学， 在HIV Tg和野生型大鼠中最先进的行为方法，以及计算策略，预计将 确定可能导致物质变革性新治疗概念的新机制假设 使用障碍（SUD）在艾滋病毒的设置，并将建立关键资源的神经艾滋病毒领域将作出 通过SCORCH数据协调中心公开提供。

Summary The abuse of stimulants such as methamphetamine (METH) exacerbates the deleterious effects of HIV infection. Here, we will carry out single nucleus RNA-Seq with the goal of identifying cell types and cell states that are pivotal in the effects of HIV and chronic methamphetamine (METH) self-administration on key brain regions relevant to the effects of persistent HIV infection and METH use disorder in HIV transgenic (Tg) rats, which harbor a non-replicating HIV-1 transgene and express chronic low-levels of multiple HIV-1 proteins. The occasional but limited use of a drug is clinically distinct from escalated drug use, which is characterized by the emergence of chronic compulsive drug-seeking and taking. Thus, we will use an established, state-of-the-art paradigm of voluntary intravenous drug self-administration under long access (LgA) conditions that leads to escalated (compulsive) METH intake in comparison to self-administration under short access (ShA) conditions, which leads to a moderate and stable pattern of METH intake. The paradigm of escalated drug intake under LgA conditions is highly relevant to the human substance use disorder (SUD) as 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 METH in this paradigm display increased compulsivity, neuroinflammation, and neural injury. The project will address the following vexing question about persistent HIV infection in the CNS: what are the cell types and cell states that drive neuroinflammation, neurodegeneration, and compulsive METH abuse in the setting of HIV that can reveal the pathogenic mechanisms behind neuroHIV disease progression, virus expression and persistence? The overarching hypothesis behind the present project is that the exploration of the gene regulatory network at the single cell level will elucidate key mechanisms that underlie the effects of HIV and METH abuse and their detrimental interactions on neuroHIV disease progression, virus expression, and virus persistence and will indicate novel therapeutic targets for neuroinflammation, neurodegeneration, and compulsivity to take METH. To test this hypothesis, we will use a validated systems biology strategy for the reconstruction and interrogation of genome-wide gene regulatory networks to identify the gene network dysregulations associated with the effects of HIV, compulsive METH use, and their interactions at single cell resolution gene profiling by single nucleus RNA-Seq. 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 is expected to identify novel mechanistic hypotheses that may lead to transformative new therapeutic concepts for substance use disorder (SUD) in the HIV setting, and will establish key resources for the neuroHIV field to be made publicly available through the SCORCH data coordination center.