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：
10405624
负责人：
Paul J. Kenny
金额：
$ 264.12万
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/10405624
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 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 Persons 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.

项目概要本申请是响应 RFA-D-21-019 提交的：单细胞阿片类药物反应 HIV 背景 (SCORCH) 计划扩展：慢性阿片类药物的 CNS 数据生成，甲基苯丙胺和/或可卡因接触可能会感染人类免疫缺陷病毒 (HIV)。大脑中的非神经元细胞，特别是小胶质细胞，这些细胞充当 HIV感染对非神经元和神经元的功能有缺失作用。细胞，包括位于大脑中与认知、情感和动机相关的部位的细胞。众所周知，受艾滋病毒影响的相同大脑部位可以调节阿片类药物和其他药物的作用。各类成瘾药物，阿片类药物使用障碍 (OUD) 在 HIV 感染者中更为普遍此外，HIV 感染和 OUD 是相互影响的。每种情况都可能加剧感染 EcoHIV 的小鼠的严重程度，一种针对 CD4+ T 细胞、巨噬细胞和小胶质细胞的改良 HIV 毒株，概括了接受抗逆转录病毒治疗的个体慢性艾滋病毒感染的主要病理生物学特征 (ART)。在这里，我们将利用最先进的单细胞测序 (scSeq)、分子、细胞测序。和行为方法来定义艾滋病毒和阿片类药物之间的细胞类型特异性相互作用在特定目标 I 中，我们优化了 EcoHIV 感染小鼠的大脑。我们的实验室已经建立了自我管理程序，用于 EcoHIV- 然后我们将检查 EcoHIV 感染对阿片类药物表达的影响。线下，我们将研究海洛因消费对成瘾相关行为的影响。 EcoHIV 感染小鼠中与 HIV 相关的认知异常的表达最后，将检查 ART 对人类神经认知障碍（HIV-NCI）的影响。 EcoHIV 感染者中成瘾和 HIV-NCI 相关行为异常的表达在特定目标 II 中，我们将对与阿片类药物成瘾相关的大脑区域进行 scSeq HIV-NCI，从没有静脉注射阿片类药物史的 EcoHIV 小鼠中收集我们将研究 ART 对这些小鼠 scSeq 模式的影响。 scSeq 数据将被挖掘来识别大脑中被 EcoHIV 感染的细胞，并确定哪些细胞在未使用阿片类药物和未使用阿片类药物的情况下对 EcoHIV 表现出最强的转录反应使用阿片类药物经历的小鼠进行原位杂交和免疫组织化学验证。在具体目标 III 中，我们将确定关键发现并优先考虑候选基因。使用体内 CRISPR-Cas9 靶向细胞类型和大脑中 Aim II 中确定的优先基因 CRISPR 切割优先基因对表达的影响。 EcoHIV 感染小鼠中与成瘾和 HIV-NCI 相关的行为异常的情况将这项创新的研究计划可能会产生根本性的新见解。 HIV 感染和阿片类药物之间与疾病相关的相互作用。