Human Endocrine Cell Development

人类内分泌细胞发育

• 批准号: 8814212
• 负责人: James M Wells
• 金额: $ 45.5万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-07 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8814212
• 关键词: Adult; Affect; BHLH Protein; Bees; Beta Cell; Birth; C-Peptide; Case Study; Cell Lineage; Cell Maturation; Cell physiology; Cells; Columnar Epithelium; Communication; Communities; Complex; Data; Development; Diabetes Mellitus; Embryo; Endocrine; Engraftment; Enteral; Enterocytes; Enteroendocrine Cell; Epithelial; Epithelium; Future; Genomic approach; Glucose; Goblet Cells; Hormones; Human; Human Development; In Vitro; Insulin; Insulin-Dependent Diabetes Mellitus; Intestinal Hormones; Intestines; Intravenous infusion procedures; Islets of Langerhans; Lead; Letters; Link; Mediating; Methods; Modeling; Molecular; Mus; Mutation; Non-Insulin-Dependent Diabetes Mellitus; Organoids; Pancreas; Pancreatic Hormones; Paneth Cells; Pathway interactions; Patients; Phenotype; Population; Production; Property; Research; Research Personnel; Role; Staging; Stem cells; Structure of beta Cell of islet; System; Testing; Therapeutic; Time; Tissues; absorption; base; blood glucose regulation; cell type; diabetic; embryonic stem cell; endocrine pancreas development; functional genomics; human embryonic stem cell; improved functioning; in vivo; induced pluripotent stem cell; intestinal epithelium; knock-down; mouse Neurog3 protein; novel; paracrine; response; small hairpin RNA; three dimensional structure; transcription factor

DESCRIPTION (provided by applicant): In mice, the basic Helix-loop-helix transcription factor Neurogenin 3 (NEUROG3) is essential for development of the endocrine lineage in both the pancreas and the intestines. Humans born with NEUROG3 mutations suffer from congenital loss of enteroendocrine cells (enteric anendocrinosis) 1 but surprisingly had normal C- peptide levels and tolerate intravenous infusion of glucose, suggesting that NEUROG3 function is not required for pancreatic beta-cell development in humans. Our new preliminary data supports this conclusion. This application with explore the cause for the apparent lack of a pancreatic endocrine phenotype in humans. We will determine if endocrine pancreas development occurred due to a partial loss of NEUROG3 activity, or alternatively NEUROG3 function is not required development of human endocrine pancreas. Identifying the role of NEUROG3 is critical since it is considered a therapeutic linchpin for generating pancreatic endocrine cells from human ES cells or via neogenesis from adult cell types. Given the rarity of tissues from patients harboring NEUROG3 mutations, it has been impossible to mechanistically investigate the basis for this difference in intestinal versus pancreatic endocrine cell development. To enable such a developmental study, we have established a culture system whereby human embryonic and induced pluripotent stem cells (collectively called PSCs) are efficiently differentiated into eithe pancreatic or intestinal tissue in vitro. Our novel method for PSC-derived intestinal tissue generates an epithelium with all of the major functioning cell types of the gut, including enteroendocrine cells. Moreover we have been able to knock down NEUROG3 in the intestinal epithelium resulting in nearly complete loss of human enteroendocrine cells. We will use development of pancreatic and intestinal endocrine cells from human PSCs as a model to identify NEUROG3-dependent and independent pathways that regulate human endocrine pancreas development. We will first determine the impact of NEUROG3 levels on pancreatic and intestinal endocrine development from human PSCs in vitro. We will identify the molecular basis for the differential requirement for NEUROG3 between the gut and pancreas by looking at the effects of NEUROG3 levels on known targets and novel pathways. Lastly we will investigate the impact of NEUROG3 levels on pancreatic and intestinal cell function in vitro and in vivo. Together, our proposed aims constitute an unprecedented functional study of NEUROG3 during human pancreas and intestinal development. Our studies will unambiguously determine if NEUROG3 is required for the development of pancreatic versus intestinal endocrine cells and if it is not required, identify the molecular pathways that compensate for absence of NEUROG3.

描述（由申请人提供）：在小鼠中，基本的helix -环-螺旋转录因子Neurogenin 3 （NEUROG3）对于胰腺和肠道内分泌谱系的发育至关重要。患有NEUROG3突变的人患有先天性肠内分泌细胞缺失（肠性内分泌失调）1，但令人惊讶的是，他们的C-肽水平正常，并能耐受静脉输注葡萄糖，这表明人类胰腺β细胞的发育并不需要NEUROG3的功能。我们新的初步数据支持这一结论。本应用将探讨人类胰腺内分泌表型明显缺乏的原因。我们将确定内分泌胰腺的发育是否由于神经g3活性的部分丧失而发生，或者神经g3功能不是人类内分泌胰腺发育所必需的。确定NEUROG3的作用至关重要，因为它被认为是从人类胚胎干细胞或通过成人细胞类型的新生生成胰腺内分泌细胞的治疗关键。鉴于患者组织中携带NEUROG3突变的罕见性，不可能从机制上研究肠道和胰腺内分泌细胞发育差异的基础。为了进行这样的发育研究，我们建立了一个培养系统，使人类胚胎和诱导多能干细胞（统称为PSCs）在体外有效地分化为胰腺或肠道组织。我们对psc来源的肠道组织的新方法产生了一个包含肠道所有主要功能细胞类型的上皮，包括肠内分泌细胞。此外，我们已经能够敲除肠上皮中的NEUROG3，导致人类肠内分泌细胞几乎完全丧失。我们将使用来自人类psc的胰腺和肠道内分泌细胞的发育作为模型，以确定调节人类胰腺内分泌发育的神经g3依赖和独立途径。我们将首先确定NEUROG3水平对体外人类PSCs胰腺和肠道内分泌发育的影响。我们将通过观察NEUROG3水平对已知靶点和新途径的影响，确定肠道和胰腺对NEUROG3需求差异的分子基础。最后，我们将在体外和体内研究NEUROG3水平对胰腺和肠细胞功能的影响。总之，我们提出的目标是对NEUROG3在人类胰腺和肠道发育过程中的功能进行前所未有的研究。我们的研究将明确确定胰腺和肠内分泌细胞的发育是否需要NEUROG3，如果不需要，则确定补偿NEUROG3缺失的分子途径。