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.

概括滥用甲基苯丙胺 (METH) 等兴奋剂会加剧艾滋病毒的有害影响感染。在这里，我们将进行单核 RNA-Seq，目的是识别细胞类型和细胞状态 HIV 和慢性甲基苯丙胺 (METH) 自我给药对关键大脑的影响至关重要与艾滋病毒转基因（Tg）大鼠中持续艾滋病毒感染和冰毒使用障碍的影响相关的区域，它含有非复制型 HIV-1 转基因，并长期表达低水平的多种 HIV-1 蛋白。这偶尔但有限地使用药物在临床上与逐步增加的药物使用不同，后者的特点是出现慢性强迫性寻药和吸毒现象。因此，我们将使用一个成熟的、最先进的在长通路（LgA）条件下自愿静脉注射药物自我给药的范例，导致与短时间（ShA）条件下的自我管理相比，METH 摄入量增加（强迫性），这导致冰毒摄入量适度且稳定。药物摄入量递增的范式 LgA 状况与人类物质使用障碍 (SUD) 高度相关，因为它模拟了所有 7 个标准《精神疾病诊断与统计手册》(DSM)-IV 中针对药物成瘾的规定以及 11 项标准中的 7 项在 DSM-V 中。我们发现，HIV Tg 大鼠在这种范式中自我施用 METH 后，显示增加强迫症、神经炎症和神经损伤。该项目将解决以下令人烦恼的问题中枢神经系统中持续的艾滋病毒感染：驱动神经炎症的细胞类型和细胞状态是什么？神经退行性变和艾滋病毒背景下强迫性滥用冰毒可以揭示致病原因神经艾滋病毒疾病进展、病毒表达和持久性背后的机制？首要的本项目背后的假设是探索单细胞的基因调控网络水平将阐明艾滋病毒和冰毒滥用及其有害影响的关键机制神经艾滋病毒疾病进展、病毒表达和病毒持久性之间的相互作用，并将表明新的神经炎症、神经退行性变和强迫性服用冰毒的治疗目标。为了测试这个假设，我们将使用经过验证的系统生物学策略来重建和询问全基因组基因调控网络，以确定与效应相关的基因网络失调 HIV、强迫性冰毒使用以及它们在单细胞分辨率基因分析中的相互作用 RNA测序。总体而言，这一跨学科协作提案整合了单细胞水平转录组学， HIV Tg 和野生型大鼠中最先进的行为方法和计算策略预计将确定新的机制假设，这些假设可能会导致物质的变革性新治疗概念艾滋病毒使用障碍（SUD），并将为神经艾滋病毒领域建立关键资源通过 SCORCH 数据协调中心公开可用。