Investigating the Role of Microglia in Methamphetamine Use Disorder

研究小胶质细胞在甲基苯丙胺使用障碍中的作用

• 批准号: 10610124
• 负责人: Samara Jo Vilca
• 金额: $ 4.78万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2023-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10610124
• 关键词: Abstinence; Adolescence; Adult; Affect; Attenuated; Behavior; Behavioral; Behavioral Paradigm; Bioinformatics; Brain; Brain region; CSF1R gene; Cell Nucleus; Cells; Cellular Morphology; Chromatin; Chronic; Complex; Data; Data Analyses; Development; Disease; Drug usage; Elderly; Epigenetic Process; FDA approved; Fostering; Gene Expression; Genes; Genetic Transcription; Goals; Home; Immune; Inflammatory; Intravenous; Learning; Libraries; Life; Longevity; Maintenance; Mediating; Mental disorders; Mentors; Methamphetamine; Methamphetamine use disorder; Microglia; Modeling; Molecular; Molecular Analysis; Morphology; Persons; Pharmaceutical Preparations; Phase; Positioning Attribute; Predisposition; Psychological reinforcement; Public Health; RNA; RNA library; Recurrent disease; Regulation; Relapse; Research; Research Personnel; Role; Self Administration; Stimulus; Subcutaneous Injections; Substance Use Disorder; Techniques; Testing; Therapeutic; Training; United States; Work; bioinformatics pipeline; brain cell; career; chromatin modification; clinically relevant; craving; disorder later incidence prevention; drug craving; drug of abuse; drug seeking behavior; early life stress; epigenome; experience; inflammatory marker; inhibitor; insight; maternal separation; methamphetamine exposure; methamphetamine use; mouse model; multiple omics; neuroinflammation; novel; pre-doctoral; response; reward processing; tenure track; transcriptome; transcriptome sequencing; transcriptomics

ABSTRACT Substance use disorder is a chronic relapsing disease that is characterized by repeated drug use despite negative consequences. Despite extensive efforts into examining the underlying molecular mechanisms of methamphetamine use disorder, there are currently no FDA approved therapeutics to treat this debilitating disease. Repeated methamphetamine (METH) use induces long-term gene expression changes in brain regions associated with reward processing and drug-seeking behavior, and recent evidence suggests that METH- induced neuroinflammation may also be involved in behavioral and molecular responses to the drug. Microglia, the resident immune cells in the brain, are principal drivers of neuroinflammatory responses, yet the role of these cells in the regulation of METH-related behaviors is poorly understood. In this proposal, I will examine microglial gene expression dynamics during METH-taking and seeking using a mouse model of intravenous methamphetamine self-administration (METH IVSA) and multi-omic molecular analyses. Preliminary data in Aim 1 (F99) demonstrate that microglia respond to METH-exposure by inducing neuroinflammatory gene expression and, accordingly, changing their cellular morphology. Additionally, since our preliminary results suggest that depletion of microglia attenuates drug-seeking after prolonged abstinence, I propose to further investigate the transcriptional and epigenetic changes in microglia in response to METH-taking and during METH craving and seeking. To this end, microglia will be isolated from METH IVSA for RNA sequencing analysis and chromatin profiling using CUT&Tag (F99). For this, I will be trained to isolate microglia for RNA and chromatin extraction, as well as the downstream bioinformatic pipelines required for data analysis. For Aim 2 (K00), I will identify a postdoctoral mentor with whom to study the effect of early life stress on microglia that contribute to later life susceptibility to psychiatric and substance use disorders. These studies will sharpen our understanding of the microglial transcriptome and epigenome during METH reinforcement and how these cells influence METH-taking and seeking behavior, while also fostering my development into a postdoctoral scholar, and ultimately for a successful career as an independent investigator in an academic tenure-track position.

抽象的 药物滥用障碍是一种慢性复发性疾病，其特点是尽管已经服用药物但仍反复使用药物 负面后果。尽管人们付出了巨大的努力来研究潜在的分子机制 甲基苯丙胺使用障碍，目前尚无 FDA 批准的疗法来治疗这种使人衰弱的疾病 疾病。重复使用甲基苯丙胺（METH）会导致大脑区域的长期基因表达变化 与奖励处理和寻求药物行为有关，最近的证据表明，METH- 诱导的神经炎症也可能与药物的行为和分子反应有关。小胶质细胞, 大脑中的常驻免疫细胞是神经炎症反应的主要驱动因素，但这些细胞的作用 人们对细胞对冰毒相关行为的调节知之甚少。在这个提案中，我将研究小胶质细胞 使用小鼠静脉注射冰毒模型服用和寻找冰毒期间的基因表达动态 甲基苯丙胺自我给药（METH IVSA）和多组学分子分析。 Aim 中的初步数据 1 (F99) 证明小胶质细胞通过诱导神经炎症基因表达来响应冰毒暴露 并相应地改变它们的细胞形态。此外，由于我们的初步结果表明 小胶质细胞的耗竭会减弱长期禁欲后的药物寻求，我建议进一步研究 小胶质细胞响应吸食冰毒以及渴望冰毒期间的转录和表观遗传变化 寻求。为此，将从 METH IVSA 中分离出小胶质细胞用于 RNA 测序分析和染色质 使用 CUT&Tag (F99) 进行分析。为此，我将接受培训以分离小胶质细胞以提取 RNA 和染色质， 以及数据分析所需的下游生物信息学管道。对于目标 2 (K00)，我将确定一个 博士后导师与他一起研究早期生活压力对小胶质细胞的影响，这有助于以后的生活 对精神疾病和药物滥用疾病的易感性。这些研究将加深我们对 冰毒强化过程中​​的小胶质细胞转录组和表观基因组以及这些细胞如何影响冰毒服用 并寻求行为，同时也促进我发展成为一名博士后学者，并最终获得一个 作为一名学术终身职位的独立调查员，取得了成功的职业生涯。