Single Cell Transcriptomic and Epigenomic Dissection of Opioid and Cocaine Responses in HIV

摘要 物质使用障碍（SUD）是一种常见的和使人衰弱的疾病，其特征是强迫性使用一种 成瘾物质，无法控制这种物质的使用，以及出现戒断症状 在没有物质的情况下。尽管我们对发生在各种环境中的变化的理解有了很大的进步， 在成瘾/SUD的背景下，对细胞类型的多样性的理解有限， 这些人脑区域内细胞的基因表达谱，以及暴露于成瘾物质 如阿片类药物和可卡因影响这些细胞的分子特性和功能。超过50%的艾滋病毒- 受感染的患者经历统称为HIV相关神经认知的神经障碍 障碍（HAND），范围从无症状的神经认知障碍到严重残疾 痴呆艾滋病毒感染者使用成瘾物质与免疫力下降有关。 功能，增加的神经炎症和神经元损伤，以及HAND的恶化。在此，我们寻求 直接剖析SUD/HIV感染/HAND效应对不同细胞的共同和不同分子基础 类型的系统分析，解剖，计算集成和实验验证其 跨个体、脑区域和细胞类型的转录、表观基因组和遗传签名。目标1：我们 使用遗传、表观基因组和转录图谱，在 单细胞（sc）水平使用2,800个样本，每个样本10,000个细胞;这些样本跨越7个大脑区域 （前额叶皮层，背侧核，腹侧被盖区，背侧纹状体，杏仁核， 海马），两种测定（scRNA，scATAC），四种表型组（SUD+/HIV+，SUD+/HIV-，SUD-/HIV+， SUD-/HIV-），共200例受试者（每个表型组50例）。目标2：我们整合这些数据集， 预测驱动基因，调控区域，变体和途径，以及细胞类型和大脑区域， 他们行动。目标3：我们在分子水平上验证高优先级的发现，并在功能上验证最高优先级的发现。 优先靶点调节体内可卡因自我给药小鼠模型中成瘾行为的能力。 由此产生的数据集将通过提供详细的治疗方法， 目标，以及它们预计会发生作用的具体条件。

Abstract Substance use disorder (SUD) is a common and debilitating condition characterized by compulsive use of an addictive substance, inability to control the use of this substance, and the emergence of withdrawal symptoms in the absence of the substance. Despite great advances in our understanding of changes that occur in various brain regions in the context of addiction/SUD, there is a limited understanding of the diversity of cell types and gene expression profiles of cells within these human brain regions, and how exposures to addictive substances such as opioids and cocaine influence the molecular properties and functions of these cells. Over 50% of HIV- infected patients experience neurological disorders collectively termed HIV-Associated Neurocognitive Disorders (HAND), which ranges from the asymptomatic neurocognitive impairment to severely disabling dementia. The use of addictive substances by HIV-infected individuals has been linked to diminished immune function, increased neuroinflammation and neuronal injury, and exacerbation of HAND. Here, we seek to directly dissect the common and distinct molecular bases of SUD/HIV infection/HAND effects on distinct cell types by systematic profiling, dissection, computational integration, and experimental validation of their transcriptional, epigenomic, and genetic signatures across individuals, brain regions, and cell types. Aim 1: We use genetic, epigenomic, and transcriptional profiles, generating a total of ~28 million genome-wide maps at the single-cell (sc) level using 2,800 samples with 10,000 cells per sample; these span 7 brain regions (prefrontal cortex, nucleus accumbens, ventral tegmental area, dorsal striatum, insula, amygdala, hippocampus), two assays (scRNA, scATAC), four phenotypic groups (SUD+/HIV+, SUD+/HIV-, SUD-/HIV+, SUD-/HIV-), and a total of 200 subjects (50 in each phenotypic group). Aim 2: We integrate these datasets to predict driver genes, regulatory regions, variants, and pathways, and the cell types and brain regions where they act. Aim 3: We validate high-priority findings at the molecular level, and functionally validate the highest priority targets’ ability to modulate addictive behaviors in mouse models of in vivo cocaine self-administration. The resulting datasets will help guide the search for new therapeutics, by providing detailed therapeutic targets, and the specific conditions where they are predicted to act.