Human Endocrine Cell Development

人类内分泌细胞发育

• 批准号: 8295786
• 负责人: NOAH Freeman SHROYER
• 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 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8295786
• 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; 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. PUBLIC HEALTH RELEVANCE: Glucose homeostasis involves the complex activity of a number of hormones produced by the endocrine cells of the pancreas and intestine. Communication between the intestinal and pancreatic endocrine cells is central to glucose homeostasis. Derangement of this system can lead to loss of beta-cell mass and/or function and result in Diabetes Mellitus. Due to the central role of pancreatic and intestinal endocrine cells i glucose homeostasis, understanding the origin and function of these cells has been the focus for current and future diabetic therapies. For example, pharmacologic manipulation of the incretin response has been an effective approach in improving the function of pancreatic beta-cells in patients with Type 2 diabetes. The pancreatic hormone insulin has been used for decades to maintain glucose homeostasis in patients with Type 1 diabetes, and production of beta-cells from stem cells continues to be a focus for many researchers as a future therapy. Studies in mice suggest that NEUROG3 is a therapeutic linchpin to generate pancreatic endocrine cells from ES cells or via neogenesis from adult cell types. This proposal will unambiguously determine if NEUROG3 is, or is not, required for both beta-cell and intestinal enteroendocrine development in HUMANS. In either case, these studies will have broad impact in the diabetes research community by determining the relevance of NEUROG3 and by identifying compensatory, alternative pathways involved in human beta-cell development.

描述（由申请人提供）：在小鼠中，基本的螺旋-环-螺旋转录因子神经原素 3 (NEUROG3) 对于胰腺和肠道中内分泌谱系的发育至关重要。出生时携带 NEUROG3 突变的人类患有先天性肠内分泌细胞缺失（肠内分泌细胞缺乏症）1，但令人惊讶的是，他们的 C 肽水平正常，并且能够耐受静脉输注葡萄糖，这表明 NEUROG3 功能并不是人类胰腺 β 细胞发育所必需的。我们的新初步数据支持了这一结论。该应用探索了人类明显缺乏胰腺内分泌表型的原因。我们将确定内分泌胰腺发育是否由于 NEUROG3 活性部分丧失而发生，或者 NEUROG3 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 ES 细胞或通过成体细胞类型新生产生胰腺内分泌细胞的治疗关键。鉴于携带 NEUROG3 突变的患者组织非常稀有，因此不可能从机制上研究肠道与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，使人胚胎干细胞和诱导多能干细胞（统称为 PSC）在体外有效分化为胰腺或肠道组织。我们针对 PSC 来源的肠道组织的新方法可生成具有肠道所有主要功能细胞类型（包括肠内分泌细胞）的上皮。此外，我们已经能够敲除肠上皮细胞中的 NEUROG3，导致人类肠内分泌细胞几乎完全丧失。我们将使用人类 PSC 的胰腺和肠内分泌细胞发育作为模型，以确定调节人类内分泌胰腺发育的 NEUROG3 依赖和独立途径。我们将首先在体外确定 NEUROG3 水平对人 PSC 胰腺和肠道内分泌发育的影响。我们将通过研究 NEUROG3 水平对已知靶点和新途径的影响，确定肠道和胰腺对 NEUROG3 差异需求的分子基础。最后，我们将研究 NEUROG3 水平对体外和体内胰腺和肠细胞功能的影响。总之，我们提出的目标构成了 NEUROG3 在人类胰腺和肠道发育过程中前所未有的功能研究。我们的研究将明确确定 NEUROG3 是否是胰腺内分泌细胞和肠内分泌细胞发育所必需的，如果不需要，则确定补偿 NEUROG3 缺失的分子途径。 公众健康相关性：葡萄糖稳态涉及胰腺和肠道内分泌细胞产生的多种激素的复杂活性。肠道和胰腺内分泌细胞之间的通讯对于葡萄糖稳态至关重要。该系统的紊乱可能导致 β 细胞质量和/或功能丧失，并导致糖尿病。由于胰腺和肠内分泌细胞在葡萄糖稳态中的核心作用，了解这些细胞的起源和功能一直是当前和未来糖尿病治疗的重点。例如，对肠促胰岛素反应进行药理学操作是改善 2 型糖尿病患者胰腺 β 细胞功能的有效方法。几十年来，胰腺激素胰岛素一直被用来维持 1 型糖尿病患者的葡萄糖稳态，而利用干细胞生产 β 细胞作为未来的治疗方法仍然是许多研究人员关注的焦点。小鼠研究表明，NEUROG3 是从 ES 细胞或通过成体细胞类型新生产生胰腺内分泌细胞的治疗关键。该提案将明确确定 NEUROG3 是否是人类 β 细胞和肠道内分泌发育所必需的。无论哪种情况，这些研究都将通过确定 NEUROG3 的相关性并确定人类 β 细胞发育中涉及的补偿性替代途径，对糖尿病研究界产生广泛影响。