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.

酗酒在年轻人中很常见。过量饮酒影响先天 通过改变基因表达和分子途径来调节肠道和大脑中的免疫信号，从而导致 酒精使用障碍（澳元）。酗酒调节的肠脑轴逆行地与 神经炎症和小胶质细胞激活。另一方面，顺行酗酒可能会导致 通过“小胶质细胞-肠道轴”，神经炎症介导的肠道菌群失调。以神经免疫系统为目标是 开发和重新利用有效药物疗法的新途径，具有临床相关性和高度 重要的。小胶质细胞中表达的烟碱 α7 乙酰胆碱受体 (7AChR) 调节 通过胆碱能抗炎途径产生神经炎症。我们发表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 巨噬细胞和小胶质细胞中的功能性 α7AChR（无效）。用 GTS-21 刺激无效的 α7AChR (α7AChR 激动剂），减少巨噬细胞中 LPS 诱导的 TNFα 释放，并用 α7AChR 敲低 α7AChR 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 摄入大量乙醇 (EtOH) 的小鼠患有肠道菌群失调，并且持续表现出显着差异 -多样性随着普雷沃特拉科科共生比例的增加而增加。采取这些场所 我们共同假设，暴食乙醇暴露会导致神经炎症和肠道菌群失调，因为 促炎细胞因子的释放，导致小胶质细胞活化并增加小胶质细胞的表达 7AChR（无效），这些因素的生物信息学分析可以建立它们之间的函数关系。 在体内，利用 PLX-5622（小胶质细胞耗竭试剂）破坏小胶质细胞和小胶质细胞 α7AChR， 小胶质细胞 α7AChR 条件敲除 (α7AChRcKO) 小鼠将分别阐明 暴食乙醇暴露后的小胶质细胞和小胶质细胞 α7AChR。 GTS-21的药理学干预 vivo 将逆转乙醇诱导的小胶质细胞激活、神经炎症和肠道菌群失调的影响，包括 7AChR。为了检验这些假设，我们提出了两个具体目标。目标 1 是表征激活 使用 C57BL/6J (B6) 小鼠和暴饮乙醇暴露后的小胶质细胞、神经炎症和肠道菌群失调 生物信息学工具。目标 2 是阐明小胶质细胞 α7AChR 的参与以及小胶质细胞在酒精- 使用 PLX-5622 和 B6 和 α7AChRcKO 小鼠调节神经炎症和肠道菌群失调 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。 R21 项目的成功完成将把观察肠脑轴的范式转变为 了解和药理学靶向暴饮暴食乙醇后的小胶质细胞-肠轴 将使我们能够开发一个完整的 R01 项目，以针对发育和发育过程中的神经免疫信号传导 重新利用针对饮酒者的有效药物疗法。