Mechanisms of Enteric Neuron Diversification

肠神经元多样化的机制

基本信息

批准号：
10670612
负责人：
E Michelle SOUTHARD-SMITH
金额：
$ 6.33万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-16 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10670612
关键词：
Achalasia Address Affect Alleles Animals Biological Assay Blood flow CCRL2 gene Cell Line Cells Chick Chromatin Chronic Complement Congenital Megacolon Constipation Data Defect Development Disease Disease model Distal Embryo Enteral Enteric Nervous System Epitopes Essential Genes Event Excision Ganglia Gastrointestinal Diseases Gastrointestinal Motility Gastroparesis Gene Expression Genes Genetic Transcription Health In Situ Individual Intestinal Diseases Intestinal Motility Intestines Knowledge Lead Life Liquid substance Mediating Megacolon Motion Mus Mutant Strains Mice Mutation Neural Crest Neuroglia Neuronal Differentiation Neurons Patients Phase Play Population Protein Isoforms Proteins Reaction Regulation Regulatory Element Role Signal Pathway Small Interfering RNA Sorting - Cell Movement Testing Transposase United States Validation Western Blotting Work antibody detection base cell motility chromatin remodeling cohesion comparative design directed differentiation experimental study fetal lentivirally transduced liquid chromatography mass spectrometry migration motility disorder mouse model mutant nerve stem cell nervous system development neurogenesis neuron development novel therapeutics nutrient absorption progenitor regenerative relating to nervous system single-cell RNA sequencing tool transcription factor transcriptomics

项目摘要

Normal gastrointestinal (GI) motility is an essential prerequisite for nutrient absorption, fecal elimination and overall health. Nearly a quarter of the United States population is affected by intestinal disorders that lead to abnormal GI motility, chronic constipation and other functional bowel disorders. Greater understanding of the mechanisms that regulate differentiation of enteric neural progenitors (ENPs), which form the neurons and glia of the enteric nervous system (ENS), are needed to understand how the normal complement of functional enteric neurons within the intestine is generated. Sox10 is an essential transcription factor that functions in the neural crest derived progenitors that generate the ENS. Defects in Sox10 in patients and mice cause aganglionosis of the distal intestine leading to megacolon. Recent studies of Sox10 mutant mice have identified pronounced alterations of the ratios of different enteric neuron types in proximal innervated bowel of these animals that are accompanied by abnormal intestinal transit and motility. Although Sox10 is expressed in ENPs, these deficiencies among enteric neurons were unexpected because Sox10 is extinguished as neurons begin to differentiate, although its expression is sustained in enteric glia. The results suggest that Sox10 has greater roles in ENS development than simply promoting migration of ENPs during initial phases when the fetal gut is first colonized by progenitors. In the proposed analysis we will test the overarching hypothesis that Sox10 action in ENPs orchestrates transcriptional networks and chromatin accessibility setting in motion a regulatory cascade that orchestrates diversity of enteric neuron subtypes. In Aim 1 we will test the hypothesis that the mutant alleles of Sox10 alter enteric neuron ratios by disrupting transcriptional hierarchies in ENPs using single cell RNA sequencing (scRNASeq). In Aim 2 we will examine the hypothesis that Sox10 mutants disrupt chromatin accessibility in developing ENS lineages. Integration of information from these studies and validation of interacting gene effects in neural crest cultures will distinguish between potential developmental mechanisms that generate the normal repertoire of enteric neuron subtypes and will facilitate efforts to direct differentiation of enteric neurons for treatment of GI disease.

正常胃肠道（GI）运动是营养吸收，消除粪便和整体健康的必要先决条件。美国近四分之一的人口受到导致异常胃肠道运动，慢性便秘和其他功能性肠道疾病的肠道疾病的影响。需要对调节肠神经祖细胞（ENP）分化的机制有更深入的了解，这些机制形成了肠神经系统（ENS）的神经元和神经胶质（ENS），以了解如何产生肠内功能性肠神经元的正常补体。 Sox10是一个基本的转录因子，它在产生ENS的神经rest衍生的祖细胞中起作用。 Sox10的患者和小鼠的缺陷导致远端肠道病，导致大巨麦龙。对Sox10突变小鼠的最新研究已经确定了这些动物近端神经支配的肠中不同肠神经元类型的比率的明显改变，这些肠伴随着异常的肠道传播和运动。尽管SOX10在ENP中表达，但肠神经元中的这些缺陷是出乎意料的，因为Sox10在神经元开始分化时被熄灭，尽管其表达在肠神经胶质中持续。结果表明，Sox10在ENS的发展中具有更大的作用，而不是简单地促进胎儿肠道首次由祖细胞定殖时的ENP迁移。在提出的分析中，我们将测试总体假设，即ENPS中的Sox10动作在运动中精心策划转录网络和染色质可及性设置，一种调节性级联，可策划肠神经元亚型的多样性。在AIM 1中，我们将通过使用单细胞RNA测序（SCRNASEQ）破坏ENPS中的转录层次来改变Sox10的突变等位基因改变肠神经元的比率的假设。在AIM 2中，我们将研究以下假设：Sox10突变体在开发ENS谱系中破坏了染色质的可及性。来自这些研究的信息的整合以及神经rest培养物中相互作用基因效应的验证将区分产生肠神经元亚型正常曲目的潜在发育机制，并将促进肠道神经元以治疗GI疾病的努力。