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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要本申请是为了回应 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 数据生成曝光。我们组建了一支优秀的研究团队，在艾滋病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）相结合，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性地自我注射可卡因。通过这种方式，我们将识别 HIV 感染的细胞类型和大脑部位。和可卡因相互作用，加剧艾滋病毒感染对大脑的负面影响，并导致感染者中持续存在可卡因使用障碍。为了实现这一目标，我们开发了一个扩展对小鼠进行静脉注射可卡因自我给药程序，可引发类似强迫的反应可卡因能够抵抗负面结果（偶然产生的有毒足部电击）。我们将利用 EcoHIV，这是一种经过修饰的艾滋病病毒毒株，可以感染传统免疫功能正常的小鼠。铬单电池多组 ATAC 和基因表达测定以及多重抗误差荧光原位杂交（MERFISH）将用于识别大脑中的可卡因和/或 HIV 反应细胞，具有前所未有的细胞和空间分辨率。我们将使用可卡因死后脑组织来证实我们的主要发现- 有经验的艾滋病毒感染者。我们的目标将通过以下四个具体目标来实现。在目标 1，我们将描述对照小鼠和感染 HIV 的小鼠的强迫性可卡因摄入量，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 和多组分析来识别那些表现出最强大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细的空间解析图受可卡因和/或艾滋病毒的影响。在目标 4 中，我们将使用死后人脑来确认我们的主要发现组织。这个高度创新、协作和多学科的研究计划有望产生关于艾滋病毒感染和可卡因使用之间与疾病相关的相互作用的基本新见解。