Single nucleus gene expression in moderate and compulsive drug self-administration in a rodent model of HIV

HIV啮齿动物模型中度和强迫性自我给药的单核基因表达

基本信息

批准号：
10592330
负责人：
PIETRO P SANNA
金额：
$ 131.89万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
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：
2022-03-15 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10592330
关键词：
Address Adopted Alzheimer&apos s Disease Animal Model Astrocytes Automobile Driving Behavior Behavioral Paradigm Binding Brain region Cell Nucleus Cells Central Nervous System Chronic Clinical Computing Methodologies DSM-IV Data Data Coordinating Center Development Disease Disease Progression Drug Addiction Drug usage Freezing Gene Expression Genes Genomics Goals HIV HIV Infections HIV-1 Head Human Impairment Intake Intravenous Lymphocyte Macrophage Malignant Neoplasms Methamphetamine Methamphetamine use disorder Methods Microglia Modeling Molecular Nerve Degeneration Neurons Neuropathogenesis Neuropsychology Neurosciences Pathogenicity Pathway Analysis Patients Pattern Persons Pharmaceutical Preparations Proteins Rat Transgene Rattus Recording of previous events Recreation Regulation Regulator Genes Research Personnel Resolution Resources Rodent Model Sampling Self Administration Substance Use Disorder Synapses Systems Biology Testing Therapeutic Tissues Transcript Transgenes Transgenic Organisms Validation Virus cell type comorbidity drug mechanism experience gene network gene regulatory network genetic signature genome-wide high throughput analysis improved methamphetamine abuse methamphetamine effect methamphetamine use nerve injury neural neuroAIDS neuroinflammation new therapeutic target novel novel therapeutics public health relevance reconstruction single cell analysis single nucleus RNA-sequencing stimulant abuse targeted treatment therapeutic target transcriptomics validation studies

项目摘要

Summary The abuse of stimulants such as methamphetamine (METH) exacerbates the deleterious effects of HIV infection. Here, we will carry out single nucleus RNA-Seq with the goal of identifying cell types and cell states that are pivotal in the effects of HIV and chronic methamphetamine (METH) self-administration on key brain regions relevant to the effects of persistent HIV infection and METH use disorder in HIV transgenic (Tg) rats, which harbor a non-replicating HIV-1 transgene and express chronic low-levels of multiple HIV-1 proteins. The occasional but limited use of a drug is clinically distinct from escalated drug use, which is characterized by the emergence of chronic compulsive drug-seeking and taking. Thus, we will use an established, state-of-the-art paradigm of voluntary intravenous drug self-administration under long access (LgA) conditions that leads to escalated (compulsive) METH intake in comparison to self-administration under short access (ShA) conditions, which leads to a moderate and stable pattern of METH intake. The paradigm of escalated drug intake under LgA conditions is highly relevant to the human substance use disorder (SUD) as it models all 7 of the criteria for drug addiction in the Diagnostic and Statistical Manual of Mental Disorders (DSM)-IV and 7 of the 11 criteria in the DSM-V. We showed that HIV Tg rats self-administering METH in this paradigm display increased compulsivity, neuroinflammation, and neural injury. The project will address the following vexing question about persistent HIV infection in the CNS: what are the cell types and cell states that drive neuroinflammation, neurodegeneration, and compulsive METH abuse in the setting of HIV that can reveal the pathogenic mechanisms behind neuroHIV disease progression, virus expression and persistence? The overarching hypothesis behind the present project is that the exploration of the gene regulatory network at the single cell level will elucidate key mechanisms that underlie the effects of HIV and METH abuse and their detrimental interactions on neuroHIV disease progression, virus expression, and virus persistence and will indicate novel therapeutic targets for neuroinflammation, neurodegeneration, and compulsivity to take METH. To test this hypothesis, we will use a validated systems biology strategy for the reconstruction and interrogation of genome-wide gene regulatory networks to identify the gene network dysregulations associated with the effects of HIV, compulsive METH use, and their interactions at single cell resolution gene profiling by single nucleus RNA-Seq. Overall, this collaborative interdisciplinary proposal integrating single cell level transcriptomics, state-of-the-art behavior methods in HIV Tg and wild-type rats, and computational strategies is expected to identify novel mechanistic hypotheses that may lead to transformative new therapeutic concepts for substance use disorder (SUD) in the HIV setting, and will establish key resources for the neuroHIV field to be made publicly available through the SCORCH data coordination center.

概括滥用诸如甲基苯丙胺（甲基苯丙胺）等兴奋剂加剧了艾滋病毒的有害作用感染。在这里，我们将执行单核RNA-Seq，其目的是识别细胞类型和细胞状态这是艾滋病毒和慢性甲基苯丙胺（甲基苯丙胺）对关键大脑的自我给药的关键与持续性HIV感染和甲基甲基苯丙胺障碍在HIV转基因（TG）大鼠中的影响相关的区域，具有非复制的HIV-1转基因和表达多种HIV-1蛋白的慢性低水平。这偶尔但使用有限的药物使用在临床上与使用升级的药物使用不同，该药物的特征是慢性强迫毒品寻求和服用的出现。因此，我们将使用已建立的最先进的在长期进入（LGA）条件下的自愿静脉药物自我管理的范例，导致与在短期访问（SHA）条件下自我管理相比，升级（强迫性）甲基甲基苯丙胺摄入量，这导致了中等稳定的甲基甲基苯丙胺摄入模式。升级的药物摄入量的范式 LGA条件与人类药物使用障碍（SUD）高度相关，因为它模拟了所有7个标准在精神障碍（DSM）的诊断和统计手册中进行吸毒 - IV和11个标准中的7个在DSM-V中。我们表明，在这种范式显示中，艾滋病毒TG大鼠自我管理的甲基强迫性，神经炎症和神经损伤。该项目将解决以下关于中枢神经系统中持续的HIV感染：驱动神经炎症的细胞类型和细胞状态是什么，在HIV的情况下，神经变性和强迫性甲基滥用，可以揭示致病性 Neurohiv疾病进展，病毒表达和持久性背后的机制？总体本项目背后的假设是对单个细胞的基因调节网络的探索级别将阐明艾滋病毒和甲基苯丙胺滥用及其有害的影响的关键机制关于神经hiv疾病进展，病毒表达和病毒持久性的相互作用，并将表明新颖神经炎症，神经变性和强迫性的治疗靶标。测试这个假设，我们将使用经过验证的系统生物学策略进行重建和审问全基因组基因调节网络，以识别与影响相关的基因网络失调艾滋病毒，强迫性甲基的使用及其在单细胞分辨率分析的相互作用。 RNA-seq。总体而言，这个合作的跨学科建议整合了单细胞级转录组学，艾滋病毒TG和野生型大鼠的最先进的行为方法，预计计算策略将有望确定可能导致实质性变革性新的治疗概念的新型机械假设在HIV环境中使用障碍（SUD），并将为神经hiv领域建立关键资源通过烧焦的数据协调中心公开获得。