MODELING ENTERIC NERVOUS SYSTEM DEVELOPMENT AND HIRSCHSPRUNG'S DISASE IN HUMAN PLURIPOTENT STEM CELLS

人类多能干细胞中肠神经系统发育和先天性巨结肠疾病的建模

• 批准号: 9219722
• 负责人: LORENZ P. STUDER
• 金额: $ 60.31万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9219722
• 关键词: Activities of Daily Living; Address; Adult; Affect; Animals; Autonomic nervous system; Biological Assay; Brain; Cell Therapy; Cell Transplantation; Cell model; Cells; Chemicals; Chick Embryo; Coculture Techniques; Colon; Communities; Complex; Congenital Abnormality; Congenital Disorders; Congenital Megacolon; Defect; Derivation procedure; Development; Disease; Disease model; Distal; Endothelin B Receptor; Endothelin B-2 Receptor; Engraftment; Enteral; Enteric Nervous System; Esophageal Atresia; Excision; Functional disorder; GDNF gene; Gastroparesis; Genetic; Genotype; Health; Hormones; Human; Hypertrophic Pyloric Stenosis; Immigration; In Vitro; Intestines; Irritable Bowel Syndrome; Lead; Life; Live Birth; Megacolon; Modeling; Molecular; Monitor; Mus; Mutant Strains Mice; Nervous System Physiology; Neural Crest; Neuroglia; Neurons; Neurotransmitters; Obstruction; Organoids; Patients; Pepstatins; Peristalsis; Pharmaceutical Preparations; Pharmacotherapy; Pluripotent Stem Cells; Pre-Clinical Model; Preclinical Drug Evaluation; Research; Role; Savings; Staging; Structure; Symptoms; Systems Biology; Technology; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Transplantation; Validation; Work; Workplace; base; cell type; disease phenotype; drug discovery; graft function; human embryonic stem cell; improved; in vivo; induced pluripotent stem cell; innovation; loss of function mutation; migration; mouse model; mutant; nervous system development; nervous system disorder; novel; novel therapeutics; optogenetics; prospective; receptor; repaired; screening; tool

SUMMARY The human enteric nervous system (ENS) is a complex network of ~500 million neurons derived from the enteric neural crest (ENC) and essential for gastro-intestinal (GI) function. ENS defects are the cause of many common human disorders including gastroparesis, irritable bowel syndrome (IBS), hypertrophic pyloric stenosis. Hirschsprung's disease (HD) is a severe congenital defect characterized by the lack of ENS precursors in the distal portions of the gut referred to as aganglionosis. Aganglionosis results in the lack of proper peristalsis causing functional colonic obstruction ("megacolon") Development of the human ENS is poorly understood given the lack of accessible tissue. The use of human pluripotent stem cells (hPSCs) represent an novel and unique strategy to access human ENS development and to model and potentially treat HD. Very recent work from our group demonstrates the feasibility of generating ENC from human PSCs and sets the stage for using the technology for developing drug and cell based strategies in treating HD and potentially other ENS disorders. The current application builds on such exciting preliminary work to pursue three specific aims: In Aim 1, we will establish conditions to characterize and manipulate the broad repertory of neuronal subtypes and enteric glia from hPSCs and to test their function in innovative co-culture assays with known target cells. In Aim 2, we will apply ENC differentiation technology to model HD in hPSCs and to validate current and identify novel candidate HD drugs. In Aim 3, we will optimize hPSC-based cell therapy approaches in a mouse model of HD and perform mechanistic studies that define the critical parameters for in vivo rescue of HD animals. Our study is unique as it represents the first effort to reliably recreate and study early human ENS lineages in vitro. The collaborators assembled for this proposal have complementary expertise in hPSCs, ENS biology, chemical screening and cell transplantation. The ENS is a structure that is not sufficiently studied by others despite its important contribution to human health. Our study has the potential to directly yield novel drug and cell based treatments for HD. The work sets the stage and develops the technology necessary for the broader community to study ENS development and ENS function in human health and disease.

概括 人类肠神经系统（ENS）是一个复杂的网络，由约5亿个神经元衍生而来 肠神经rest（ENC），对于胃肠道（GI）功能必不可少。 ENS缺陷是许多原因的原因 常见的人类疾病，包括胃轻瘫，肠易激综合征（IBS），肥厚性幽门疾病 狭窄。 Hirschsprung病（HD）是一种严重的先天性缺陷，其特征是缺乏ENS 肠道远端的前体被称为Aganglionisos。 Aganglionisos导致缺乏 适当的蠕动引起功能性结肠阻塞（“巨型巨龙”） 鉴于缺乏可进入的组织，人类ENS的发展知之甚少。使用 人类多能干细胞（HPSC）代表了一种新颖而独特的策略，可以访问人类ENS 开发，建模和可能治疗高清。我们小组的最新工作证明了 从人类PSC产生ENC的可行性，并为使用该技术开发奠定了基础 基于药物和细胞的策略治疗高清和潜在的其他ENS疾病。当前的应用程序 建立在如此令人兴奋的初步工作的基础上，以追求三个具体目标： 在AIM 1中，我们将建立表征和操纵神经元亚型的广泛曲目的条件 来自HPSC的肠神经胶质细胞，并测试其在与已知靶细胞的创新共培养测定中的功能。在 AIM 2，我们将采用ENC DICANITION技术在HPSC中建模并验证电流并确定 新型候选高清药物。在AIM 3中，我们将在小鼠模型中优化基于HPSC的细胞疗法 高清并进行机械研究，以定义体内拯救HD动物的关键参数。 我们的研究是独一无二的 体外。为此建议组装的合作者在HPSC，ENS Biology， 化学筛查和细胞移植。 ENS是一个没有其他人充分研究的结构 尽管它对人类健康有重要贡献。我们的研究有可能直接产生新的药物和 基于细胞的HD治疗。该作品设定了舞台，并开发了更广泛的技术 社区研究ENS发展和ENS在人类健康和疾病中的功能。