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

基本信息

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
项目类别：
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

项目摘要

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，过量饮酒会影响先天。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（AUD）与酗酒调节的肠脑轴逆行相关。另一方面，酗酒可能会导致神经炎症和小胶质细胞激活。神经炎症介导的肠道菌群失调，通过“小胶质细胞-肠道轴”来靶向神经免疫系统。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在小胶质细胞中表达的烟碱 α7 乙酰胆碱受体 (7AChR) 具有重要的调节作用。我们发表的数据和其他研究表明，神经炎症通过胆碱能抗炎途径发生。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 (α7AChR) 刺激无效的 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。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变化。我们共同发现，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) 小鼠将分别阐明暴饮 EtOH 暴露后小胶质细胞和小胶质细胞 α7AChR 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，包括为了检验这些假设，我们提出了两个具体目标：表征 α7AChR 的激活。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中的作用。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。