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: 10705750
• 负责人: SULIE L. CHANG
• 金额: $ 18.22万
• 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/10705750
• 关键词: 16S ribosomal RNA sequencing; 3-Dimensional; Abbreviations; Acetylcholine; Address; Affect; Agonist; Alcohol consumption; Alcohols; Anti-Inflammatory Agents; Bacteria; Bacteroidetes; Bioinformatics; Brain; Cholinergic Receptors; Communication; 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; Leaky Gut; Length; Lipopolysaccharides; Liver diseases; Macrophage; Mediating; Meta-Analysis; Microglia; Modeling; Molecular; Mus; Neuroimmune; Neuroimmune system; Nitrogen; Outcome; Oxygen; Pathway interactions; Periodicity; Persons; 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; binge drinking; bioinformatics tool; chemokine; cholinergic; clinically relevant; conditional knockout; cytokine; cytotoxic; deter alcohol use; dysbiosis; genetic information; genetic manipulation; glial activation; gut dysbiosis; gut microbiome; gut microbiota; gut-brain axis; high reward; high risk; in vivo; in vivo Model; innovation; knock-down; microbiome; microbiome research; microbiota; neuroinflammation; pathogenic bacteria; pharmacologic; systemic inflammatory response; translational study; 16S ribosomal RNA sequencing; 3-Dimensional; Abbreviations; Acetylcholine; Address; Affect; Agonist; Alcohol consumption; Alcohols; Anti-Inflammatory Agents; Bacteria; Bacteroidetes; Bioinformatics; Brain; Cholinergic Receptors; Communication; 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; Leaky Gut; Length; Lipopolysaccharides; Liver diseases; Macrophage; Mediating; Meta-Analysis; Microglia; Modeling; Molecular; Mus; Neuroimmune; Neuroimmune system; Nitrogen; Outcome; Oxygen; Pathway interactions; Periodicity; Persons; 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; binge drinking; bioinformatics tool; chemokine; cholinergic; clinically relevant; conditional knockout; cytokine; cytotoxic; deter alcohol use; dysbiosis; genetic information; genetic manipulation; glial activation; gut dysbiosis; gut microbiome; gut microbiota; gut-brain axis; high reward; high risk; in vivo; in vivo Model; innovation; knock-down; microbiome; microbiome research; microbiota; neuroinflammation; pathogenic bacteria; pharmacologic; 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.

暴饮暴食在年轻人中非常普遍。过度饮酒会影响先天 通过改变基因表达和分子途径，肠道和大脑中的免疫信号传导 酒精使用障碍（AUD）。屈光酒精调节的肠脑轴逆行，与 神经炎症和小胶质细胞激活。另一方面，狂饮，同类可能会导致 神经炎症介导的肠癌通过“小胶质细胞轴”。靶向神经免疫系统是 开发和重新利用有效药物治疗的新途径，它在临床上相关且高度 重要的。烟碱α7乙酰胆碱受体（7ACHR），在小胶质细胞调节中表达 神经炎症通过胆碱能抗炎途径。我们发布的数据和其他研究表明 脂多糖（LPS）诱导的炎症释放促炎性细胞因子并上调非 - 巨噬细胞和小胶质细胞中的功能性7ACHR（徒劳）。用GTS-21刺激徒劳的7ACHR（7ACHR） 激动剂），减少巨噬细胞中LPS诱导的TNF释放，并用7ACHR敲除7ACHR 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 给予暴饮暴食（ETOH）的小鼠患有肠道营养不良，并不断暴露于 多样性，以及prevotellaceae家族的共生比例增加。采用这些前提 我们共同假设暴力ETOH暴露会导致神经炎症和肠道疾病障碍。 促炎细胞因子的释放，这会导致小胶质细胞激活和小胶质细胞的表达增加 7ACHR（徒劳的）和对这些因素的生物信息学分析可能会建立它们之间的功能关系。 在体内，使用PLX-5622（小胶质细胞耗竭试剂）和小胶质细胞的破坏和小胶质细胞7ACHR和 小胶质细胞7ACHR条件敲除（7ACHRCKO）小鼠将阐明 小胶质细胞和小胶质细胞暴露后的小胶质细胞。 GTS-21的药物干预 体内将逆转ETOH诱导的对小胶质细胞激活，神经炎症和肠道营养不良的影响，涉及 7ACHR。为了检验这些假设，我们有两个具体的目标。目标1是表征激活的 使用C57BL/6J（B6）小鼠暴露后，小胶质细胞，神经炎症和肠道营养不良，并 生物信息学工具。目标2是阐明小胶质细胞7ACHR和小胶质细胞在酒精中的作用的参与。 通过使用B6和7ACHRCKO小鼠使用PLX-5622和 GTS-21。我们具有很高临床相关性的创新项目可能具有高风险，但很可能很高的奖励。 该R21项目的成功完成应将范式从观察肠道轴转移到 在暴饮暴食中暴露于EtOH和 将使我们能够开发一个全长R01项目，以针对开发和 重新利用酒精饮用者的有效药物治疗。