Genetic and extrinsic mechanisms governing early enteric nervous system development

控制早期肠神经系统发育的遗传和外在机制

• 批准号: 10581603
• 负责人: Rosa A Uribe
• 金额: $ 39.81万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-27 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10581603
• 关键词: Address; Affect; Agreement; Anterior; Biological Assay; Biological Models; Candidate Disease Gene; Cell Cycle; Cells; Chagas Disease; Chromosome Mapping; Collecting Cell; Colon; Colonic Aganglionosis; Complex; Computer Models; Congenital Megacolon; Constipation; Coupled; Data; Data Set; Defect; Destinations; Development; Distal; Embryo; Embryonic Development; Ensure; Enteral; Enteric Nervous System; Environment; Equilibrium; Esophageal achalasia; Event; Eye; Foundations; Future; Ganglia; Gastrointestinal tract structure; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Genomics; Goals; Hindgut; Homeostasis; Hormone secretion; Human; Immigration; In Situ; Infiltration; Intestinal Motility; Intestinal Obstruction; Knowledge; Lead; Length; Mammals; Maps; Mitosis; Mitotic; Modeling; Molecular; Muscle; Nervous System control; Neuroglia; Neuronal Differentiation; Neurons; Optics; Pathway interactions; Pattern; Peristalsis; Play; Population; Positioning Attribute; Process; Proliferating; Research; Research Proposals; Resolution; Role; Series; Signal Pathway; Signal Transduction; Speed; System; Testing; Therapeutic Studies; Time; Tissue Engineering; Tissues; Tretinoin; Tube; Water; Zebrafish; cell fate specification; confocal imaging; effective therapy; enteric neuropathy; experimental study; gastrointestinal; gene regulatory network; genetic signature; gut colonization; in utero; in vivo; in vivo imaging; innovation; intestinal barrier; loss of function; migration; nerve stem cell; nervous system development; nervous system disorder; neural; neurodevelopment; neurogenesis; neuron development; neuronal patterning; novel therapeutics; optogenetics; programs; spatiotemporal; stem cells; transcription factor; transcriptomics; translational therapeutics

Resident between the muscle walls of the entire gastrointestinal (GI) tract, the enteric nervous system (ENS) consists of a series of interconnected neurons and glia, numbered in the hundreds of millions. The ENS controls essential gut functions, such as peristalsis, water balance and intestinal barrier homeostasis. The ENS is derived from enteric neural progenitors (ENPs) that migrate into the developing gut tube during embryogenesis and differentiate into enteric neurons or glia. Disruption in ENS formation results in the congenital condition Hirschsprung disease (HSCR), in which variable regions of the GI lack ENS—the most common form of HSCR presents along the distal colon, also known as colonic aganglionosis. The underlying cellular mechanisms that ENPs utilize to migrate into and spatially position along the gut tube, as well as genetic programs they execute to differentiate into enteric neurons have not been well studied in vivo, therefore limiting our knowledge of how the ENS manifests. The overall goal is to expand foundational knowledge of the genes utilized to execute the complex mechanisms necessary for ENS formation, with an eye for informing downstream translational therapeutic studies. In this proposal, we utilize zebrafish embryos due to their genetic conservation with humans, the ease of viewing their external development and for their optical transparency. Building off of single-cell transcriptomic data sets generated from ENP cells collected during their early neurogenesis along the gut tube, Aim 1 will examine a hypothesis that the spatial arrangement of newly uncovered ENP transcriptional subpopulations predict future enteric neuron placement and terminal differentiation along the gut tube. In agreement with and extending observations in mammalian models, we have recently discovered that Retinoic Acid (RA) signaling is critical globally during early steps of zebrafish ENS development; however, how RA signaling autonomously influences ENS ontogenesis in vivo is not well understood in any system to date. Aim 2 will investigate a hypothesis that the RA pathway autonomously controls ENP differentiation states and migration patterns along the gut tube using cutting edge single-cell transcriptomics, optogenetics and in vivo imaging. We will also test a mechanistic model in Aim 2 that candidate transcription factors function intrinsically downstream of RA in ENPs to govern ENS formation, thereby expanding our understanding of the ENS gene regulatory network. Aim 3 will use genetic modulation of the cell cycle, quantitative in vivo imaging and cell tracking test a cellular mechanistic model that ENPs couple proliferation with migration to dictate proper enteric neuron patterning in the gut downstream of the RA pathway. The results of these aims will significantly increase our knowledge of the genetic, molecular and cellular underpinnings of ENP development and early ENS creation and they will provide a new mechanistic framework for studying these developmentally important cells in vivo.

整个胃肠道（GI）的肌肉壁之间的居民，肠神经系统 （ENS）由一系列相互连接的神经元和神经胶质组成，编号为数亿美元。这 ENS控制着必不可少的肠道功能，例如蠕动，水平衡和肠道障碍稳态。 ENS源自肠神经祖细胞（ENP），在 胚胎发生并分化为肠神经元或神经胶质。在Ens形成中的破坏导致 先天性状况Hirschsprung病（HSCR），其中gi的可变区域缺乏ENS，最多 HSCR沿远端结肠的常见形式，也称为结肠aganglionisos。这 ENP用于迁移到肠道管的空间位置的基本细胞机制， 除了遗传程序外，他们执行以分化为肠神经元的遗传程序还没有很好地研究 因此，体内，限制了我们对ENS如何表现的知识。总体目标是扩展 用于执行ENS所需的复杂机制的基因的基础知识 形成，着眼于告知下游翻译理论研究。在这个建议中，我们 由于其与人类的遗传保护，利用斑马鱼的胚胎，易于观察其外部 开发及其光学透明度。建立在生成的单细胞转录组数据集中 从沿肠管的早期神经发生期间收集的ENP细胞中，AIM 1将检查A 假设新发现的ENP转录亚群的空间排列预测 沿肠管的未来肠神经元放置和终末分化。与 扩展了哺乳动物模型中的观察结果，我们最近发现视黄酸（RA） 在斑马鱼ENS发育的早期步骤中，信号在全球范围内至关重要。但是，ra发信号如何 迄今为止，在任何系统中都无法很好地理解体内的ENS个体发生。 AIM 2意志 研究RA途径自主控制ENP分化状态和 使用尖端单细胞转录组学，光遗传学和IN沿着肠管的迁移模式 体内成像。我们还将在AIM 2中测试一个机械模型，该模型候选转录因子起作用 在ENP中，RA的本质下游以管理Ens形成，从而扩大了我们对 ENS基因调节网络。 AIM 3将使用细胞周期的遗传调节，体内定量 成像和细胞跟踪测试一个细胞机械模型，该模型将夫妇增殖与迁移到 在RA途径的肠道中指示适当的肠神经元图案。这些结果 目标将显着提高我们对ENP遗传，分子和细胞基础的了解 开发和早期的创造，它们将为研究这些提供新的机械框架 在体内发育重要的细胞。