Characterizing neurogenic progenitors in the adult intestine

成人肠道神经源性祖细胞的特征

基本信息

批准号：
9895033
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
项目类别：
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/9895033
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

项目摘要

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.

项目概要肠神经系统（ENS）通过调节所有器官对人类健康至关重要胃肠道 (GI) 功能方面，最显着的是先天性或后天性肠道蠕动异常。因此，ENS 可导致严重的功能性胃肠道疾病，包括食管贲门失弛缓症、胃轻瘫、假性肠梗阻、肠易激综合征、先天性巨结肠和慢传输型便秘。众所周知，成人肠道拥有神经元祖细胞，但它们的作用和激活机制我们和其他人观察到特定实验损伤中新神经元的诞生。模型，包括肠道炎症、胃肠道感染和局灶性神经元消融后的模型。反应可以替代因损伤而失去的神经元，但它也可以产生神经增生，从而导致神经元增生。因此，肠神经发生是一把双刃剑。可以发挥有益作用，但需要进行调整以限制其后果。表明啮齿类动物的实验性结肠炎促进肠神经胶质细胞通过然而，人们对这一过程的总体目标知之甚少。是了解产后肠神经发生的机制及其在胃肠道健康中的作用为了实现这一目标，我们提出以下具体目标：（1）确定下游途径。由 5-HT4 信号传导激活并导致神经胶质细胞分化为神经元 (2) 的特征肠道中存在的神经胶质细胞亚群，并确定遗传和表观遗传变化发生在神经胶质细胞到神经元命运转换期间；（3）利用肠道的神经发生能力来治疗与先天性巨结肠症相关的神经节下移行区将有多种方法。用于实现这些目标，包括从报告小鼠中分离和培养肠神经胶质细胞；使用显性失活突变体和药理学抑制剂来确定参与的信号通路神经胶质神经发生；单细胞 RNA seq 鉴定神经胶质细胞亚群；用于神经胶质细胞到神经元细胞命运转变的活细胞成像，分析遗传和表观遗传变化；发生在该过渡期间；以及诱导 HSCR 肠道中的肠神经发生以治疗过渡区成功完成这些实验将显着增强我们对神经节减退症的理解。成人肠道神经发生的机制，提供对病理生理学的见解神经肠道疾病，确定调节体内神经发生的新靶标，为治疗神经肠道疾病提供新方法 HSCR 下神经节过渡区和其他神经肠中的肠神经元疾病，并改善肠神经元祖细胞的体外扩增，用于细胞治疗应用。