Characterizing neurogenic progenitors in the adult intestine

成人肠道神经源性祖细胞的特征

• 批准号: 10066349
• 负责人: ALLAN M GOLDSTEIN
• 金额: $ 36.33万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10066349
• 关键词: Ablation; Adult; Agonist; Birth; Cell Therapy; Cells; Chronic; Clinical; Colitis; Colon; Congenital Megacolon; Constipation; Coupled; Data; Digestive System Disorders; Disease; Distal; Dominant-Negative Mutation; Enteral; Enteric Nervous System; Epigenetic Process; Esophageal achalasia; Esthesia; Excision; Experimental Models; Functional Gastrointestinal Disorders; Functional disorder; Gastroparesis; Gene Expression; Genetic Transcription; Glial Differentiation; Goals; Health; Human; Hyperplasia; Immunity; In Vitro; Injury; Intestinal Pseudo-Obstruction; Intestines; Irritable Bowel Syndrome; Knowledge; Lead; Life; Mediating; Methodology; Modeling; Morbidity - disease rate; Mucositis; Mus; Mutation; Nervous System Physiology; Neuroglia; Neurons; Pathologic; Pathway interactions; Pharmacology; Population Heterogeneity; Process; Regulation; Reporter; Rodent; Role; Serotonin; Signal Pathway; Signal Transduction; Source; Structure; Subgroup; Surface; System; Testing; Transgenic Organisms; absorption; base; cell motility; density; design; experimental study; gastrointestinal; gastrointestinal function; gastrointestinal infection; genetic analysis; improved; in vitro Assay; in vivo; inflammatory disease of the intestine; inhibitor/antagonist; insight; live cell imaging; loss of function; motility disorder; mouse model; nerve stem cell; nervous system development; neurogenesis; novel strategies; postnatal; progenitor; relating to nervous system; response; serotonin receptor; single-cell RNA sequencing; stem cell biology; stem cells; transcriptome; transdifferentiation

Project Summary The enteric nervous system (ENS) is of fundamental importance to human health through its regulation of all aspects of gastrointestinal (GI) function, most notably gut motility. Congenital or acquired abnormalities of the ENS consequently can lead to serious functional GI disorders, including esophageal achalasia, gastroparesis, intestinal pseudo-obstruction, irritable bowel syndrome, Hirschsprung disease, and slow transit constipation. The adult intestine is known to possess neuronal progenitors, but their role and the mechanisms that activate them are unknown. We and others have observed the birth of new neurons in specific experimental injury models, including intestinal inflammation, GI infection, and following focal neuronal ablation. This neurogenic response can serve to replace neurons lost to injury, but it can also produce neuronal hyperplasia which can have significant pathologic consequences. Enteric neurogenesis is thus a double-edged sword that can be leveraged for a beneficial effect but needs to be modulated to limit its consequences. Our preliminary results suggest that experimental colitis in rodents promotes enteric glial cells to undergo a neurogenic transition via a 5-HT4-dependent pathway. However, this process is poorly understood. The overall objective of this proposal is to understand the mechanisms underlying postnatal enteric neurogenesis and its role in GI health and disease. To achieve this goal, we propose the following specific aims: (1) identify the downstream pathways that are activated by 5-HT4 signaling and lead to glial differentiation into neurons; (2) characterize the subpopulations of glial cells present in the intestine and determine the genetic and epigenetic changes that occur during the glia-to-neuron fate switch; and (3) leverage the intestine’s capacity for neurogenesis to treat the hypoganglionic transition zone associated with Hirschsprung disease. A variety of methodologies will be used to achieve these aims, including isolation and culture of enteric glia from reporter mice; in vitro assays using dominant-negative mutants and pharmacologic inhibitors to determine the signaling pathways involved in glial neurogenesis; single cell RNA seq to identify glial cell subpopulations; a dual reporter transgenic system for live cell imaging of glia-to-neuron cell fate transition; analysis of the genetic and epigenetic changes occurring during that transition; and induction of enteric neurogenesis in HSCR bowel to treat transition zone hypoganglionosis. Successful completion of these experiments will significantly enhance our understanding of the mechanisms underlying neurogenesis in the adult intestine, provide insights into the pathophysiology of neurointestinal diseases, identify new targets to modulate neurogenesis in vivo, offer novel approaches for expanding enteric neurons in the hypoganglionic transition zone of HSCR and in other neurointestinal diseases, and improve the in vitro expansion of enteric neuronal progenitors for cell therapy applications.

项目摘要 肠神经系统（ENS）通过对所有人的监管对人类健康至关重要 胃肠道（GI）功能的各个方面，最著名的是肠道运动。先天性或获得的异常 因此，ENS可以导致严重的功能性胃肠道疾病，包括食管阿萨拉氏症，胃轻瘫， 肠道伪腹结构，肠易激综合征，赫希斯普朗病和慢速过境便秘。 已知成年肠具有神经元的祖细胞，但其作用和激活的机制 他们是未知的。我们和其他人在特定的实验损伤中观察到了新神经元的诞生 模型，包括肠道注射，胃肠道感染和局灶性神经元消融。这种神经源 反应可以用来代替因受伤而失去的神经元，但它也可以产生神经元增生，可以 具有重大的病理后果。因此，肠神经发生是一把双刃剑 利用有益的效果，但需要调节以限制其后果。我们的初步结果 表明啮齿动物中的实验性结肠炎会促进肠神经胶质细胞通过A进行神经源性过渡 5-HT4依赖性途径。但是，这个过程对此很了解。该提议的总体目标 是要了解产后促进神经发生及其在胃肠道健康中的作用的机制 疾病。为了实现这一目标，我们提出以下特定目的：（1）确定下游途径 通过5-HT4信号传导激活，并导致神经元分化为神经元。 （2）表征 肠道中存在神经胶质细胞的亚群，并确定遗传和表观遗传变化 发生在神经胶质到神经元的命运开关期间； （3）利用肠道神经发生的能力 与Hirschsprung疾病相关的降节性过渡区。各种方法将是 用于实现这些目标，包括从记者小鼠中的肠神经胶质的隔离和培养；体外测定 使用显性阴性突变体和药物抑制剂来确定涉及的信号传导途径 神经神经发生；单细胞RNA SEQ鉴定神经胶质细胞亚群；双重记者转基因系统 用于神经胶质到神经元细胞命运过渡的活细胞成像；分析遗传和表观遗传变化 在过渡期间发生；和诱导HSCR肠中肠神经发生以治疗过渡区 低血管病。这些实验的成功完成将大大增强我们对 成人肠中神经发生的基础机制，提供了有关对病理生理学的见解 神经性疾病，确定在体内调节神经发生的新靶标，为您提供新的方法 在HSCR和其他神经结构性的肾小管过渡区域中扩展肠神经元 疾病，改善用于细胞治疗应用的肠神经元祖细胞的体外扩张。