Single cell transcriptomic and epigenomic changes during chronic HIV infection and cocaine self-administration

慢性艾滋病毒感染和可卡因自我给药期间的单细胞转录组和表观基因组变化

• 批准号: 10454708
• 负责人: Paul J. Kenny
• 金额: $ 355.77万
• 依托单位: ALLEN INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10454708
• 关键词: Abstinence; Archives; Autopsy; Behavior; Behavioral; Biological Assay; Brain; Brain region; Cell Nucleus; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Chromatin; Chromium; Chronic; Cocaine; Cocaine Abuse; Cocaine Dependence; Cocaine use disorder; Consumption; Corpus striatum structure; Coupled; Data; Data Set; Development; Disease; Environment; Epigenetic Process; Evaluation; Fluorescent in Situ Hybridization; Gene Expression; Gene Expression Profiling; Generations; Genes; Genetic Transcription; Goals; HIV; HIV Infections; HIV Seronegativity; HIV-1; Habenula; Human; Human immunodeficiency virus test; Image; Immunocompetent; Individual; Infection; Intake; Intervention; Intravenous; Lateral; Maps; Measures; Modeling; Molecular; Morbidity - disease rate; Mus; National NeuroAids Tissue Consortium; Neurobiology; Neurocognitive Deficit; Neuroglia; Neurology; Neurons; Neurosciences; Opioid; Outcome; Pathologic; Persons; Pharmaceutical Preparations; Positioning Attribute; Prefrontal Cortex; Procedures; Property; Punishment; RNA; Research; Research Personnel; Resistance; Resolution; Resources; Rewards; Risk; Risk Factors; Sampling; Shock; Site; Small Nuclear RNA; Technology; Testing; Tissue Donors; Tissues; Training; Validation; Volition; XCL1 gene; addiction; antiretroviral therapy; base; behavioral response; brain cell; brain tissue; cell type; cocaine exposure; cocaine self-administration; cocaine use; cohort; deep sequencing; epigenome; epigenomics; experience; foot; innovation; insight; methamphetamine exposure; mortality; multidisciplinary; nano; neuropathology; opioid exposure; overdose death; programs; response; single cell sequencing; synergism; transcriptome; transcriptome sequencing; transcriptomics; virology

PROJECT SUMMARY This application is submitted in response to RFA-D-21-019: Single Cell Opioid Responses in the Context of HIV (SCORCH) Program Expansion: CNS Data Generation for Chronic Opioid, Methamphetamine, and/or Cocaine Exposures. We have assembled an outstanding team of investigators with world-class expertise in HIV virology, addiction neurobiology, and single cell transcriptomics. We will leverage state-of-the-art single-nuclei RNA sequencing (snRNA-seq) coupled to single-nuclei epigenetics (snATAC-seq) to define the transcriptional and epigenetic landscape of neuronal and nonneuronal cells in addiction-relevant brain cells of HIV-infected mice that compulsively self-administer cocaine. In this manner we will identify cell types and brain sites in which HIV and cocaine interact to exacerbate the negative impact of HIV infection on the brain and contribute to the persistence of cocaine use disorder in infected individuals. To facilitate this goal, we have developed an extended access intravenous cocaine self-administration procedure for mice that precipitates compulsive-like response for cocaine that is resistant to negative outcome (contingently delivered noxious foot shocks). We will utilize EcoHIV, which is a modified HIV strain that can infect conventional immunocompetent mice. The Chromium Single Cell Multiome ATAC and Gene Expression assay and Multiplexed error-robust fluorescence in situ hybridization (MERFISH) will be used to identify cocaine and/or HIV-responsive cells in the brain with unprecedented cellular and spatial resolution. We will confirm our major findings using postmortem brain tissues from cocaine- experienced HIV-infected individuals. Our goals will be accomplished through the following four specific aims. In Aim 1, we will characterize compulsive-like cocaine intake in control and HIV-infected mice and determine the impact of antiretroviral treatment (ART) on cocaine response in these mice. In Aim 2, we will use snRNA-seq and Mulitome assays to identify those cells that show the most robust transcriptional and epigenetic plasticity to cocaine consumption and HIV infection. In Aim 3, we will use MERFISH to confirm and validate our major snRNA- seq and Multiome assay findings, and generate more refined spatially resolved maps of those cells most impacted by cocaine and/or HIV. In Aim 4, we will confirm our major findings using post-mortem human brain tissues. This highly innovative, collaborative and multidisciplinary program of research promises to yield fundamental new insights into disease-related interactions between HIV infection and cocaine use.

项目摘要 本申请是为了响应RFA-D-21-019：HIV背景下的单细胞阿片类药物反应而提交的 （SCORCH）计划扩展：慢性阿片类药物、甲基苯丙胺和/或可卡因的CNS数据生成 暴露。我们组建了一支优秀的研究人员团队，在艾滋病毒学方面拥有世界一流的专业知识， 成瘾神经生物学和单细胞转录组学。我们将利用最先进的单核RNA 测序（snRNA-seq）与单核表观遗传学（snATAC-seq）偶联，以定义转录和 HIV感染小鼠成瘾相关脑细胞中神经元和非神经元细胞的表观遗传景观 强迫自己服用可卡因通过这种方式，我们将确定艾滋病毒感染的细胞类型和大脑部位。 和可卡因相互作用，加剧了艾滋病毒感染对大脑的负面影响，并有助于 感染者中可卡因使用障碍的持续存在。为了实现这一目标，我们开发了一个扩展的 进入静脉内可卡因自我管理程序的小鼠，沉淀强迫样反应， 对负面结果有抵抗力的可卡因（偶然传递的有害脚电击）。我们将利用EcoHIV， 它是一种改良的HIV病毒株，可以感染常规的免疫活性小鼠。铬单电池 多组ATAC和基因表达测定以及多重错误稳健荧光原位杂交 （MERFISH）将用于识别大脑中的可卡因和/或艾滋病毒反应细胞， 空间分辨率。我们将用可卡因的死后脑组织来证实我们的主要发现- 经验丰富的艾滋病毒感染者。我们将通过以下四个具体目标来实现我们的目标。在 目的1，我们将描述控制组和HIV感染组小鼠的强迫性可卡因摄入，并确定 抗逆转录病毒治疗（ART）对这些小鼠可卡因反应的影响。在目标2中，我们将使用snRNA-seq 和Mulitome测定来鉴定那些显示出最强的转录和表观遗传可塑性的细胞， 可卡因消费和艾滋病毒感染。在目标3中，我们将使用MERFISH来确认和验证我们的主要snRNA- seq和Multiome检测结果，并生成这些细胞的更精确的空间分辨图。 可卡因和/或艾滋病毒的影响。在目标4中，我们将使用死后人脑来证实我们的主要发现 组织中这个高度创新，协作和多学科的研究计划有望产生 对艾滋病毒感染和可卡因使用之间与疾病有关的相互作用的基本新见解。