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

摘要 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，目的是鉴定细胞类型和细胞状态 它们在艾滋病毒和慢性甲基苯丙胺(冰毒)自我给药对关键大脑的影响中起关键作用 HIV转基因(TG)大鼠中HIV持续感染和冰毒使用障碍影响的相关区域， 它携带非复制的HIV-1转基因，并表达慢性低水平的多种HIV-1蛋白。这个 偶尔但有限地使用药物在临床上不同于逐步增加的药物使用，后者的特征是 慢性强迫性寻药和吸毒现象的出现。因此，我们将使用一个成熟的、最先进的 长通道(LGA)条件下自愿静脉注射药物自我给药的范例 与短期接触(SHA)条件下的自我给药相比，增加(强制)冰毒摄入量， 这导致了适度和稳定的冰毒摄入模式。中国毒品摄入量上升的范式 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)中的药物成瘾--11项标准中的第四项和第七项 在DSM-V中。我们发现，在这一范例中，HIV TG大鼠自我给予冰毒的表现增加 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： 中枢神经系统持续的HIV感染：导致神经炎症的细胞类型和细胞状态是什么， 神经退行性变，以及强迫性冰毒滥用在HIV的背景下可以揭示其致病机制 神经HIV疾病进展、病毒表达和持久性背后的机制？最重要的是 本项目背后的假设是，对单细胞基因调控网络的探索 将阐明艾滋病毒和冰毒滥用的影响及其有害的关键机制 神经HIV疾病进展、病毒表达和病毒持久性的相互作用将预示新的 治疗神经炎、神经退行性变和服用冰毒的强迫症。为了测试这一点 假设，我们将使用经过验证的系统生物学策略来重建和审问 全基因组基因调控网络识别与基因网络失调相关的影响 HIV、强迫性冰毒的使用及其在单核单细胞分辨基因图谱中的相互作用 RNA-Seq.总体而言，这一跨学科的协作提案整合了单细胞水平的转录组学， 在HIV、TG和野生型大鼠中的最新行为方法和计算策略有望 确定可能导致变革性新的物质治疗概念的新的机械假说 在艾滋病毒环境中使用障碍(SUD)，并将为将要建立的神经艾滋病毒领域建立关键资源 可通过SCOCH数据协调中心公开获取。

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.