Single cell determinants of brain in the context of viral persistence in SIV/cART/cocaine non-human primates

项目摘要 药物滥用和艾滋病毒感染并存，由于艾滋病毒感染者面临的挑战，很难进行研究 （PLWH）滥用药物的人。艾滋病毒和滥用药物的致病相互作用的主要领域之一是 大脑，这带来了额外的研究限制。为了获得关于艾滋病毒和可卡因对 大脑，我们将利用充分研究的非人类灵长类动物模型，猿猴免疫缺陷病毒（SIV）- 感染和未感染的对照恒河猴，在存在或不存在可卡因的情况下。单细胞 测序分析使得能够对特定细胞类型在健康大脑和脑组织中的作用进行细胞类型特异性分析。 功能与疾病由多学科调查员小组提出的这项建议的总体目标是， 揭示了在SIV/cART/可卡因非人类中病毒持续存在的背景下脑的单细胞决定因素 灵长类动物单细胞RNA测序和单细胞转座酶可降解性检测 染色质测序（scMultiome-seq）将在分离自前额叶皮层（PFC）的细胞上进行， 纹状体和海马。将对数据进行处理和分析，以得出细胞类型特异性 调节程序和这些程序的变化，调节灵长类动物大脑中的神经发病机制。 这些研究将为SIV/可卡因/cART在大脑中的相互作用提供独特而有价值的见解， 这也为人类的发现提供了关键的实验验证。成功完成这些 创新的单细胞研究和数据分析方法将产生单细胞转录组， 三个NIDA相关脑区PFC、纹状体和海马的细胞部分列表的表观基因组图谱。 这些结果将阐明CNS的转录组学和表观遗传学景观的新见解， 他们被艾滋病毒和可卡因的使用改变了。

PROJECT SUMMARY Drug abuse and HIV infection co-exist and are difficult to study due to challenges in people living with HIV (PLWH) who abuse drugs. One of the prime areas of pathogenic interactions of HIV and drugs of abuse is the brain, which carries additional study limitations. To obtain rigorous data on the effects of HIV and cocaine on the brain, we will utilize well-studied nonhuman primate animal model, Simian immunodeficiency virus (SIV)- infected and uninfected control rhesus monkeys, in the presence or absence of cocaine. Single cell sequencing assays enable cell type-specific analysis of the role of specific cell types in both healthy brain function and disease. The overall objective of this proposal by multidisciplinary team of investigators is to reveal the single cell determinants of brain in the context of viral persistence in SIV/cART/cocaine non-human primates. Simultaneous single-cell RNA sequencing and single-cell Assay for Transposase- Accessible Chromatin sequencing (scMultiome-seq) will be carried out on cells isolated from the prefrontal cortex (PFC), striatum, and hippocampus from each animal. Data will be processed and analyzed to derive cell type-specific regulatory programs and changes in these programs that regulate neuropathogenesis in the primate brains. These studies will provide unique and valuable insights into SIV/cocaine/cART interactions in the brain while also providing crucial experimental validation of the findings in humans. Successful completion of these innovative single-cell studies and data analyses methods will generate a single cell transcriptome and epigenome atlas of cellular part list for three NIDA-relevant brain regions, PFC, striatum, and hippocampus. These results will illuminate novel insights into transcriptomic and epigenetic landscapes of CNS and how they are altered by HIV and cocaine use.