The Unfolding Role of Microglia in Alcohol Withdrawal

小胶质细胞在酒精戒断中的作用

• 批准号: 10491273
• 负责人: John F Neumaier
• 金额: $ 16.17万
• 依托单位: SEATTLE INST FOR BIOMEDICAL/CLINICAL RES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10491273
• 关键词: Abstinence; Address; Air; Alcohol Withdrawal Delirium; Alcohol abuse; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcohols; Animals; Apoptosis; Area; Behavior; Behavior Therapy; Behavioral; Biology; Brain; Cell Death; Cells; Cellular Stress Response; Cessation of life; Characteristics; Chronic; Code; Complication; Corpus striatum structure; Cre driver; Data; Development; Enterobacteria phage P1 Cre recombinase; Ethanol; Ethanol toxicity; Exploratory/Developmental Grant; Exposure to; Filopodia; Gene Expression; Genes; Genetic Transcription; Genotype; Goals; Immune; Induction of Apoptosis; Inflammatory; Intoxication; Knock-out; Lead; Life; LoxP-flanked allele; Measures; Mediating; Medical emergency; Messenger RNA; Methods; Microglia; Microscopic; Microscopy; Modeling; Molecular; Morphology; Motivation; Mouse Protein; Mus; Negative Reinforcements; Neuroimmune; Neuronal Plasticity; Neurons; Oxidative Stress; Pathway interactions; Pharmaceutical Preparations; Process; Protein Biosynthesis; Proteins; RNA; Regulation; Relapse; Reporter; Research; RiboTag; Ribosomes; Risk; Role; Seizures; Severities; Signal Transduction; Structure; Substance Withdrawal Syndrome; Surveys; Synapses; Testing; Time; Toxic effect; Transgenic Mice; Transgenic Organisms; Translations; Withdrawal; Withdrawal Symptom; alcohol abstinence; alcohol abuse therapy; alcohol consequences; alcohol exposure; alcohol measurement; alcohol relapse; alcohol use disorder; base; cell motility; cohort; conditional knockout; cytokine; endoplasmic reticulum stress; experience; experimental study; in vivo; innovation; neuroinflammation; next generation sequencing; novel; protein expression; response; transcription factor; transcription factor CHOP; transcriptome sequencing; two photon microscopy; two-photon; vapor

Abstract Alcohol withdrawal is a medical emergency and a behavioral barrier to reducing alcohol abuse. Unlike withdrawal to other drugs, there is ample evidence that withdrawal to alcohol becomes worse after multiple cycles of alcohol consumption and abstinence, increasing the risks for serious adverse complications including seizures, delirium tremens, and death. Furthermore, the negative reinforcement inherent to cycles of alcohol abstinence and relapse to alcohol consumption complicates any behavioral interventions to reduce alcohol abuse. Many clues suggest that neuroinflammation is involved in both alcohol tolerance as well as withdrawal, but this realization has not led to new treatments for alcohol use disorder. The purpose of this proposal is to develop our hypothesis that chronic, intermittent alcohol exposure alters the way microglia, the innate immune cells that reside in the brain, modulate neuroimmune as well as neuronal synaptic signaling. In pilot experiments using RNA sequencing of ribosome-associated RNA from microglia after multiple cycles of alcohol exposure and abstinence, we found that alcohol withdrawal causes oxidative stress in microglia and activates the “unfolded protein response” (UPR), a mechanism that is involved in cellular stress responses leading to neuroinflammatory signaling and derangements in microglia function that can lead to cell death. We propose to block the full activation of the microglial UPR by knocking out CHOP, a key transcription factor involved in this pathway that was dramatically upregulated in microglia during withdrawal, using a recently developed, selective cre-driver mouse line, Tmem119-2A-Cre-ERT2/TdTomato, crossed with commercially available floxed CHOP mice to investigate this biology. We will examine how blunting UPR signaling alters the signs of alcohol withdrawal by testing if CHOP knockout alters several behavioral manifestations and reduces microglia morphological and gene expression changes associated with withdrawal. Using in vivo two-photon time lapse microscopy, we will examine if CHOP knockout from microglia changes morphology and motility associated during alcohol withdrawal. We will also test whether microglial CHOP impacts the motivation to drink alcohol in a model of voluntary alcohol drinking after multiple cycles of ethanol vapor exposure and withdrawal. Together, these experiments examine the role of neuroinflammation in the mechanisms of alcohol dependence and withdrawal from a new angle that has not been previously considered and is well matched to the R21 mechanism due to its high novelty and potential to develop new treatments.

抽象的 戒酒是医疗紧急情况，也是减少酒精滥用的行为障碍。与众不同 提取其他药物，有充分的证据表明，多次戒酒会变得更糟 饮酒和禁欲的循环，增加了严重广告并发症的风险 癫痫发作，del妄和死亡。此外，负强化继承了酒精周期 戒酒和饮酒的缓解使任何行为干预措施都复杂化以减少酒精 虐待。许多线索表明，神经炎症既参与酒精的耐受性和戒断， 但是，这种认识并未导致对酒精使用障碍的新疗法。该提议的目的是 提出我们的假设，即慢性，间歇性酒精暴露会改变小胶质细胞的方式，先天性免疫 驻留在大脑中的细胞会调节神经免疫性和神经元突触信号传导。在飞行员中 在多个酒精循环后使用核糖体相关的RNA的RNA测序 暴露和禁欲，我们发现戒酒会导致小胶质细胞中的氧化应激并激活 “展开的蛋白质反应”（UPR），这种机制参与细胞应激反应，导致 小胶质细胞功能的神经炎症信号传导和演变可能导致细胞死亡。 我们建议通过将CHOP（一个关键的转录因子）敲开小胶质细胞UPR的全部激活 使用最近的小胶质细胞中在小胶质细胞中急剧上调的这一途径，使用最近 开发的，有选择性的Cre-Driver小鼠系列TMEM119-2A-CRE-ERT2/TDTOMATO，与商业交叉 可用的flox斩波小鼠研究了这种生物学。我们将检查钝器信号的钝化如何改变 通过测试CHOP敲除改变了几种行为表现并减少的迹象 小胶质细胞的形态和基因表达与戒断有关。使用体内两光子 时间衰变显微镜，我们将检查小胶质细胞的切碎敲除是否改变形态和运动性 在戒酒期间相关。我们还将测试小胶质排骨是否影响 多次乙醇蒸气暴露和 提取。这些实验共同研究了神经炎症在酒精机制中的作用 依赖性和从以前尚未考虑的新角度提取，并且与 R21机制由于其高新颖性和开发新处理的潜力而引起的。