Neural sensing of gut bacteria

肠道细菌的神经感知

• 批准号: 10671590
• 负责人: Winston W Liu
• 金额: $ 4.98万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10671590
• 关键词: 3-Dimensional; Acute; Address; Bacteria; Bathing; Biology; Brain; Brain Stem; Calcium; Cell Communication; Cells; Central Nervous System; Chemicals; Classification; Clinical; Colon; Communication; Data; Dependence; Detection; Disease; Dissociation; Enteroendocrine Cell; Epithelial Cells; Esthesia; Flagellin; Gastroenterology; Gastrointestinal Diseases; Glutamate Transporter; Glutamates; Hormones; Human; Image; In Vitro; Ingestion; International; Intestines; Irritable Bowel Syndrome; Knock-out; Knowledge; Laboratories; Ligands; Link; Measures; Mediating; Mentors; Metabolic syndrome; Microbe; Molecular; Mus; Names; Nerve; Neurobiology; Neurosciences; Neurotransmitters; Nutrient; Obesity; Organoids; Patients; Pattern; Pattern recognition receptor; Pelvis; Peptide YY; Pharmacology; Physicians; Population; Postdoctoral Fellow; Reporting; Research Project Grants; Research Training; Role; Sacral nerve; Science; Scientist; Sensory; Signal Transduction; Societies; Specificity; Synapses; TLR5 gene; Techniques; Testing; Therapeutic; Training; Training Support; United States National Institutes of Health; Visceral; Weight Gain; burden of illness; career preparation; cellular transduction; clinical training; commensal bacteria; conditional knockout; doctoral student; extracellular; gut bacteria; gut microbiome; hormonal signals; in vivo; intestinal epithelium; microbial; microbial community; microbiome; millisecond; neural; neural circuit; neurotransmitter release; novel; optogenetics; pathogen; pharmacologic; receptor; response; sensory mechanism; skills; symposium; therapeutic target; tool

ABSTRACT The gut continuously senses resident bacteria, but how the brain recognizes these microbes remains unclear. Microbe-associated molecular patterns such as flagellin are detected by the intestinal epithelium by pattern recognition receptors such as Toll-like receptor 5. Recent reports have shown that knocking out this receptor in the gut leads to metabolic syndrome. Pattern recognition receptors are also known to be preferentially expressed on enteroendocrine cells, or electrically excitable epithelial cells traditionally thought to signal hormonally. The sponsor’s laboratory has recently discovered that some of these cells, now known as neuropod cells, are synaptically connected with vagal and pelvic nerves. My preliminary data show that the neuropod cells preferentially express Toll-like receptor 5, and that conditionally knocking out the receptor in neuropod cells leads to weight gain in mice. These data suggest that neuropod cells are critical intermediaries in bacterial signaling from gut to brain. Therefore, the central hypothesis of the research project is that neuropod cells transduce flagellin in the lumen of the colon onto the sacral nerve through a synapse. To test this, two aims are proposed: (1) to determine whether neuropod cells release glutamate in response to flagellin in vitro, and (2) to test neuropod cell transduction of flagellin onto the sacral nerve in vivo. To address these aims, state-of-the-art techniques from intestinal epithelial biology and neurobiology will be combined by the trainee. In Aim 1, acutely dissociated neuropod cells and 3-dimensional organoid cultures will be imaged for calcium activity and glutamate release in response to flagellin. In Aim 2, optogenetic and pharmacological silencing of neuropod cells in sacral nerve recordings will be used to test whether the circuit transduces bacterial signals. These studies are expected to uncover a novel mechanism for microbes to communicate with the central nervous system that can be used to develop therapeutics for patients with gastrointestinal disease. This proposal will ultimately support the training of a dual-degree MD/PhD student, in preparation for his career as an independent physician-scientist at the intersection of a clinical gastroenterology practice and a neuroscience laboratory. The training plan will also include attending conferences and participating in organizing an international society of gut-brain scientists started by his sponsor named Gastronauts. With the support of this F30, the trainee will develop the requisite skill set to transition into post-doctoral clinical and research training on the path to becoming an independent physician scientist.

抽象的 肠道持续感知常驻细菌，但大脑如何识别这些微生物仍然存在 不清楚。肠道检测到微生物相关的分子模式，例如鞭毛蛋白 上皮细胞通过模式识别受体（例如 Toll 样受体 5）进行识别。最近的报告表明 敲除肠道中的这种受体会导致代谢综合征。模式识别受体 还已知优先在肠内分泌细胞上表达，或可电兴奋 传统上认为上皮细胞发出激素信号。主办方实验室最近 发现其中一些细胞，现在被称为神经足细胞，与突触相连 迷走神经和骨盆神经。我的初步数据显示神经足细胞优先表达 Toll 样受体 5，有条件地敲除神经足细胞中的受体会导致体重增加 小鼠的增益。这些数据表明神经足细胞是细菌信号传导的关键中介 从肠道到大脑。因此，该研究项目的中心假设是神经足细胞 通过突触将结肠腔中的鞭毛蛋白转导到骶神经上。为了测试这一点， 提出了两个目标：（1）确定神经足细胞是否响应于释放谷氨酸 （2）在体内测试鞭毛蛋白到骶神经的神经足细胞转导。到 为了实现这些目标，肠上皮生物学和神经生物学的最先进技术将 由实习生组合。在目标 1 中，急性分离的神经足细胞和 3 维类器官 对培养物响应鞭毛蛋白的钙活性和谷氨酸释放进行成像。在目标 2 中， 将使用骶神经记录中神经足细胞的光遗传学和药理学沉默 测试电路是否转导细菌信号。这些研究预计将揭示一个 微生物与中枢神经系统通讯的新机制可用于 开发针对胃肠道疾病患者的治疗方法。该提案最终将得到支持 对双学位医学博士/博士生的培训，为他的独立职业生涯做好准备 临床胃肠病学实践和神经科学交叉领域的医师科学家 实验室。培训计划还将包括参加会议和参与组织 由他的赞助商“Gastronauts”发起成立了一个由肠脑科学家组成的国际协会。随着 在该 F30 的支持下，学员将培养过渡到博士后临床所需的技能 以及成为独立医师科学家之路上的研究培训。