Involvement of microglial α7AChR in binge alcohol modulation of gut dysbiosis

小胶质细胞α7AChR参与酗酒调节肠道菌群失调

• 批准号: 10527744
• 负责人: SULIE L. CHANG
• 金额: $ 22.01万
• 依托单位: SETON HALL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10527744
• 关键词: 16S ribosomal RNA sequencing; 3-Dimensional; Abbreviations; Address; Affect; Agonist; Alcohol consumption; Alcohols; Anti-Cholinergics; Anti-Inflammatory Agents; Bacteria; Bacteroidetes; Bioinformatics; Brain; Cholinergic Receptors; Communication; Control Groups; Correlation Studies; Data; Data Set; Elements; Endotoxins; Epidemic; Ethanol; Exhibits; Exposure to; Factor Analysis; Family; Feedback; Firmicutes; GTS-21; Gene Expression; Genetic; Goals; Growth; Health; Healthcare Systems; Heavy Drinking; Homeostasis; Immune signaling; Inflammation; Inflammatory; Intervention; Knockout Mice; Lead; Leaky Gut; Length; Lipopolysaccharides; Liver diseases; Mediating; Meta-Analysis; Microglia; Modeling; Molecular; Mus; Neuroimmune; Neuroimmune system; Nitrogen; Outcome; Oxygen; Pathway interactions; Periodicity; Persons; Pharmacology; Pharmacotherapy; Plasma; Probiotics; Proteobacteria; Publishing; RNA; Reagent; Regulation; Reporting; Role; Signal Transduction; Small Interfering RNA; TNF gene; Taxonomy; Testing; Therapeutic Intervention; Time; Up-Regulation; Water; alcohol exposure; alcohol use disorder; alcohol use initiation; alpha-bungarotoxin receptor; base; binge drinking; bioinformatics tool; chemokine; clinically relevant; conditional knockout; cytokine; cytotoxic; deter alcohol use; dysbiosis; genetic information; genetic manipulation; gut dysbiosis; gut microbiome; gut microbiota; gut-brain axis; high reward; high risk; in vivo; in vivo Model; innovation; knock-down; macrophage; microbiome; microbiome research; microbiota; neuroinflammation; pathogenic bacteria; systemic inflammatory response; translational study

Binge alcohol drinking is very common among the young people. Excessive alcohol consumption affects innate immune signaling in the gut and the brain by altering gene expression and molecular pathways which lead to alcohol use disorder (AUD). The binge alcohol-modulated gut-brain axis, retrogradely, is associated with neuroinflammation and microglial activation. On the other hand, binge alcohol, anterogradely, may cause neuroinflammation-mediated gut dysbiosis, via the “microglia-gut axis”. Targeting the neuroimmune system is a new avenue for developing and repurposing effective pharmacotherapies and it is clinically relevant and highly significant. The nicotinic alpha7 acetylcholine receptor (7AChR), which is expressed in microglia regulates neuroinflammation through cholinergic anti-inflammatory pathway. Our published data and other study showed that lipopolysaccharide (LPS)-induced inflammation releases pro-inflammatory cytokines and upregulates non- functional 7AChR (futile) in macrophages and microglia. Stimulation of futile 7AChR with GTS-21 (7AChR agonist), decreases LPS-induced TNF release in macrophages, and knockdown of 7AChR with 7AChR siRNA in LPS-induced macrophages abolish GTS-21 beneficial effects. Our preliminary studies demonstrated that mice given binge ethanol (EtOH) suffer with gut dysbiosis and continuously exhibit significant differences in -diversity along with an increase in commensal proportions of Prevotellaceae family. Taken these premises together, we hypothesize that binge EtOH exposure causes neuroinflammation and gut dysbiosis due to the release of proinflammatory cytokines, which lead to microglial activation and increased expression of microglial 7AChR (futile), and bioinformatic analysis of these factors could establish a functional relationship among them. In in vivo, disruption of microglia and microglial 7AChR utilizing PLX-5622 (microglia depletion reagent) and microglial 7AChR conditional knockout (7AChRcKO) mice, respectively, will elucidate the discrete roles of microglia and microglial 7AChR after binge EtOH exposure. The pharmacological intervention of GTS-21 in vivo will reverse EtOH-induced effects on microglia activation, neuroinflammation and gut dysbiosis involving 7AChR. To test these hypotheses, we have posited two specific aims. Aim 1 is to characterize activation of microglia, neuroinflammation and gut dysbiosis following binge EtOH exposure using C57BL/6J (B6) mice and bioinformatics tools. Aim 2 is to elucidate the involvement of microglial 7AChR and role of microglia in alcohol- modulation of neuroinflammation and gut dysbiosis by utilizing B6 and 7AChRcKO mice using PLX-5622 and GTS-21. Our innovative project with high clinical relevance, might have high risk, yet very likely high reward. Successful completion of this R21 project shall shift the paradigm from observing the gut-brain axis to understanding and pharmacologically targeting the microglia-gut axis following binge exposure to EtOH and would allow us to develop a full length R01 project to target the neuroimmune signaling in developing and repurposing effective pharmacotherapies for alcohol drinkers.

酗酒在年轻人中很普遍。过量饮酒影响先天性 通过改变基因表达和分子途径， 酒精使用障碍（AUD）酗酒调节的肠脑轴，逆行，与 神经炎症和小胶质细胞活化。另一方面，酗酒，顺行，可能会导致 神经炎症介导的肠道生态失调，通过“小胶质细胞-肠道轴”。靶向神经免疫系统是一种 这是开发和重新利用有效药物疗法的新途径，具有临床相关性， 显著在小胶质细胞中表达的烟碱α 7乙酰胆碱受体（NAT 7AChR）调节 通过胆碱能抗炎途径抑制神经炎症。我们公布的数据和其他研究表明， 脂多糖（LPS）诱导的炎症释放促炎细胞因子，并上调非炎症细胞因子， 巨噬细胞和小胶质细胞中的功能性β 7 AChR（无效）。用GTS-21刺激无效的β_7 AChR（β_7 AChR 激动剂），减少LPS诱导的巨噬细胞中TNF α的释放，并用β 7 AChR敲低β 7 AChR LPS诱导的巨噬细胞中的siRNA消除了GTS-21的有益作用。我们的初步研究表明 给予过量乙醇（EtOH）的小鼠患有肠道生态失调，并在以下方面持续表现出显著差异： 植物多样性沿着增加，植物比例也随之增加。把这些房子 总之，我们假设过量的乙醇暴露会导致神经炎症和肠道生态失调， 释放促炎细胞因子，导致小胶质细胞活化和小胶质细胞表达增加 AChR（futile），通过生物信息学分析，可以建立它们之间的功能关系。 在体内，利用PLX-5622（小胶质细胞耗竭试剂）和 小胶质细胞ACh 7AChR条件性敲除（ACh 7AChRcKO）小鼠，分别阐明的离散作用， 小神经胶质细胞和小神经胶质细胞α 7AChR在过量EtOH暴露后的变化。GTS-21的药理干预作用 体内将逆转EtOH诱导的对小胶质细胞活化、神经炎症和肠道生态失调的影响， 7AChR。为了验证这些假设，我们提出了两个具体目标。目的1是表征 使用C57 BL/6 J（B6）小鼠，在过量EtOH暴露后观察小胶质细胞、神经炎症和肠道生态失调， 生物信息学工具。目的二是阐明小胶质细胞AChR的参与和小胶质细胞在酒精中毒中的作用。 通过使用PLX-5622和PLX-5622利用B6和B7 AChRcKO小鼠调节神经炎症和肠道生态失调， GTS-21我们的创新项目具有高度的临床相关性，可能具有高风险，但很可能具有高回报。 这个R21项目的成功完成将使范式从观察肠-脑轴转变为 了解和靶向小胶质细胞肠轴后，狂欢暴露于乙醇和 将使我们能够开发一个完整的R 01项目，以靶向神经免疫信号在发展和 为饮酒者重新利用有效的药物疗法。