The Unfolding Role of Microglia in Alcohol Withdrawal

小胶质细胞在酒精戒断中的作用

基本信息

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
项目类别：
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

项目摘要

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.

抽象的酒精戒断是一种医疗紧急情况，也是减少酗酒的行为障碍。对其他药物的戒断，有充分的证据表明，多次戒酒后对酒精的戒断会变得更糟饮酒和戒酒的循环，增加了严重不良并发症的风险，包括此外，酒精循环固有的负面强化。戒酒和酗酒使任何减少饮酒的行为干预措施变得复杂许多线索表明神经炎症与酒精耐受和戒断有关，但这种认识并没有带来针对酒精使用障碍的新疗法。该提案的目的是。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（先天免疫）的方式存在于大脑中的细胞调节神经免疫以及神经元突触信号传导。使用多次酒精循环后小胶质细胞核糖体相关 RNA 的 RNA 测序实验暴露和戒酒，我们发现酒精戒断会引起小胶质细胞的氧化应激并激活 “未折叠蛋白反应”（UPR），一种参与细胞应激反应的机制，导致神经炎症信号传导和小胶质细胞功能紊乱可导致细胞死亡。我们建议通过敲除关键转录因子 CHOP 来阻止小胶质细胞 UPR 的完全激活使用最近的一项研究，参与了这条在戒断期间小胶质细胞中急剧上调的通路开发的选择性 cre-driver 小鼠品系 Tmem119-2A-Cre-ERT2/TdTomato，与商业杂交我们将使用 floxed CHOP 小鼠来研究这种生物学特性。通过测试 CHOP 敲除是否会改变一些行为表现并减少酒精戒断症状使用体内双光子观察小胶质细胞形态和基因表达的变化。延时显微镜，我们将检查小胶质细胞的 CHOP 敲除是否会改变形态和运动性我们还将测试小胶质细胞 CHOP 是否会影响戒酒的动机。在多次乙醇蒸气暴露循环后，在自愿饮酒模型中饮酒这些实验共同探讨了神经炎症在酒精机制中的作用。从以前没有考虑过的新角度进行依赖和退出，并且非常匹配 R21 机制因其高度新颖性和开发新疗法的潜力而受到关注。