Single cell brain transcriptome changes during chronic HIV infection and opiate use in conventional mice

传统小鼠慢性艾滋病毒感染和阿片类药物使用期间单细胞脑转录组的变化

基本信息

批准号：
10220584
负责人：
Paul J. Kenny
金额：
$ 265.47万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-15 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10220584
关键词：
AIDS dementia Abstinence Behavior Behavioral Bone Diseases Brain Brain region CD4 Positive T Lymphocytes CRISPR/Cas technology Candidate Disease Gene Cells Centers for Disease Control and Prevention (U.S.)Cessation of life Chronic Cleaved cell Clustered Regularly Interspaced Short Palindromic Repeats Cognition Cognitive Cognitive deficits Consequences of HIV Consumption DNA Data Data Set Development Diabetes Mellitus Disease Drug usage Emotions Gene Expression General Population Generations Genes Genetic Transcription Goals HIV HIV Infections HIV-1 Helper-Inducer T-Lymphocyte Heroin Human Immunocompetent Immunohistochemistry In Situ Hybridization Individual Infection Inflammation Intravenous Investigation Kidney Diseases Laboratories Liver diseases Memory impairment Microglia Molecular Morbidity - disease rate Morphine Motivation Mus Neurocognitive Deficit Neuroglia Neuronal Dysfunction Neurons Opiate Addiction Opioid Overdose Patients Pattern Pharmaceutical Preparations Population Procedures Recording of previous events Research Resolution Rewards Risk Risk Factors Self Administration Severities Site System T-Lymphocyte Technology Testing Transcription Process Viral Proteins Virus Volition addiction antiretroviral therapy brain cell cardiovascular disorder risk cell type cocaine exposure cognitive enhancement comorbidity drug of abuse experience heroin use in vivo innovation insight latent infection macrophage methamphetamine exposure mortality neuroAIDS neuron loss opioid abuse opioid exposure opioid injection opioid mortality opioid overdose opioid use opioid use disorder overdose death programs response single cell sequencing transcriptome

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. Human immunodeficiency virus (HIV) can infect nonneuronal cells in the brain, particularly microglia, with these cells acting as a reservoir of latent infection. HIV infection has deletions effects on the function of nonneuronal and neuronal cells, including those cells located in brain sites relevant to cognition, emotion and motivation. The same brain sites impacted by HIV are known to regulate the actions of opioids and other classes of addictive drugs, and opioid use disorder (OUD) is more prevalent in HIV-infected individuals than the general population. Moreover, HIV infection and OUD reciprocally interact, with each condition potentially exacerbating the severity of the other. Mice infected with EcoHIV, a modified HIV strain that targets CD4+ T cells, macrophages and microglia, recapitulates the major pathobiological features of chronic HIV infection in individuals on antiretroviral therapy (ART). Here, we will leverage state-of-the-art single cell sequencing (scSeq), molecular, cellular and behavioral approaches to define cell type-specific interactions between HIV and opioids in the brains of EcoHIV-infected mice. In Specific Aim I, we optimize the intravenous (IV) heroin self-administration procedure, already well-established in our laboratories, for use in EcoHIV- infected mice. We will then examine the effects of EcoHIV infection on the expression of opioid addiction-relevant behaviors. Conversely, we will examine the effects of heroin consumption on the expression of cognitive abnormalities in EcoHIV-infected mice relevant to HIV-associated neurocognitive impairment (HIV-NCI) in humans. Finally, will examine the effects of ART on the expression of addiction- and HIV-NCI-relevant behavioral abnormalities in EcoHIV-infected mice. In Specific Aim II, we will perform scSeq on brain regions relevant to opioid addiction and HIV-NCI, collected from EcoHIV mice with our without a history of intravenous opioid self- administration behavior. We will investigate the effects of ART on scSeq patterns in these mice. The scSeq data will be mined to identify cells in the brain infected by EcoHIV, and determine which cells show the most robust transcriptional responses to EcoHIV in opioid-naïve and opioid-experienced mice. In situ hybridization and immunohistochemistry will be used to validate key findings and prioritize candidate genes for further investigation. In Specific Aim III, we will use in vivo CRISPR-Cas9 to target prioritized genes identified in Aim II in a cell type- and brain region-specific manner. The impact of CRISPR cleavage of prioritized genes on the expression of addiction- and HIV-NCI-relevant behavioral abnormalities in EcoHIV-infected mice will be examined. This innovative program of research may yield fundamental new insights into disease-relevant interactions between HIV infection and opioid drugs.

项目摘要该应用程序是针对RFA-D-21-019的响应提交的：艾滋病毒（Scorch）计划扩展的上下文：慢性阿片类药物的CNS数据生成，甲基苯丙胺和/或可卡因暴露。人类免疫缺陷病毒（HIV）可以感染大脑中的非神经元细胞，尤其是小胶质细胞，这些细胞充当潜在感染。 HIV感染对非神经元和神经元的功能具有缺失影响细胞，包括与认知，情绪和动机有关的大脑部位中的细胞。已知受HIV影响的相同大脑部位调节阿片类药物和其他的作用在HIV感染中，添加剂类别和阿片类药物使用障碍（OUD）更为普遍个人比普通人群。此外，HIV感染和OUD相互作用，每种条件都可能加剧对方的严重程度。感染Ecohiv的小鼠，针对CD4+ T细胞，巨噬细胞和小胶质细胞的改良HIV菌株，概括了抗逆转录病毒疗法中慢性HIV感染的主要病原体特征（艺术）。在这里，我们将利用最新的单细胞测序（SCSEQ），分子，细胞以及定义艾滋病毒和阿片类药物之间特定细胞类型相互作用的行为方法生态感染的小鼠的大脑。在特定目的I中，我们优化了静脉（IV）海洛因在我们的实验室中已经建立了良好的自我管理程序，用于Ecohiv- 感染的小鼠。然后，我们将检查生态感染对Opoid表达的影响与成瘾相关的行为。相反，我们将研究海洛因消耗的影响与HIV相关的生态感染小鼠认知异常的表达人类的神经认知障碍（HIV-NCI）。最后，将研究艺术对成瘾 - 与HIV-NCI相关的行为异常的表达老鼠。在特定的目标II中，我们将对与阿片类药物成瘾相关的大脑区域进行SCSEQ和 HIV-NCI，从我们的Ecohiv小鼠中收集，我们没有静脉注射自我的病史行政行为。我们将研究艺术对这些小鼠SCSEQ模式的影响。 SCSEQ数据将被开采以鉴定被EcoHIV感染的大脑中的细胞，并确定哪些细胞在阿片类药物中显示出对EcoHIV的最强转录反应，阿片类药物经验的小鼠。原位杂交和免疫组织化学将用于验证关键发现并确定候选基因以进行进一步研究。在特定的目标III中，我们将使用体内CRISPR-CAS9来靶向在细胞类型和大脑中AIM II中鉴定的优先基因区域特定方式。优先基因的CRISPR分裂对表达的影响成瘾 - 与HIV-NCI相关的行为异常在Ecohiv感染的小鼠中将是检查。这项创新的研究计划可能会产生基本的新见解艾滋病毒感染与阿片类药物之间的相关疾病相互作用。