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蛋白3（Neurog3）对于胰腺和肠中内分泌谱系的发展至关重要。患有神经3突变的人类遭受肠内内分泌细胞的先天性丧失（肠形分泌症）1，但令人惊讶的是，糖肽水平正常，耐受静脉输注葡萄糖，这表明人类胰腺β细胞发育不需要Neurog3功能。我们的新初步数据支持了这一结论。该应用具有探讨显然缺乏人类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神经3活性的部分损失而发生，或者不需要开发人类内分泌胰腺的Neurog3功能。识别Neurog3的作用至关重要，因为它被认为是一种用于从人ES细胞产生胰腺内分泌细胞或通过成人细胞类型的新生成的治疗素丁丁。鉴于具有Neurog3突变的患者的组织稀有性，因此无法机械地研究肠道内分泌细胞发育与胰腺内分泌细胞发育的这种差异的基础。为了实现这样的发展研究，我们建立了一个培养系统，在该系统中，人类胚胎和诱导的多能干细胞（统称称为PSC）在体外有效地分化为胰腺或肠道组织。我们对PSC衍生的肠道组织的新方法产生了上皮，其中包括肠道的所有主要功能细胞类型，包括肠内分泌细胞。此外，我们已经能够在肠上皮中击倒Neurog3，从而几乎完全丧失了人类肠内分泌细胞。我们将使用人类PSC的胰腺内分泌细胞和肠内内分泌细胞的开发作为模型，以识别调节人内分泌胰腺发育的Neurog3依赖性和独立的途径。我们将首先确定Neurog3水平对体外人类PSC的胰腺内分泌发育的影响。我们将通过查看Neurog3水平对已知靶标和新途径的影响来确定肠道和胰腺之间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。我们提出的目标共同构成了人类胰腺和肠发展过程中神经3的前所未有的功能研究。我们的研究将明确确定胰腺内分泌细胞的发展是否需要Neurog3，如果不需要肠道内分泌细胞，则确定补偿缺乏Neurog3的分子途径。 公共卫生相关性：葡萄糖稳态涉及胰腺和肠内内分泌细胞产生的许多激素的复杂活性。肠道内分泌细胞和胰腺内分泌细胞之间的通信对于葡萄糖稳态至关重要。该系统的危险会导致β细胞质量和/或功能的损失，并导致糖尿病。由于胰腺和肠内内分泌细胞I葡萄糖稳态的核心作用，了解这些细胞的起源和功能一直是当前和未来的糖尿病疗法的重点。例如，肠降凝素反应的药理操作是改善2型糖尿病患者胰腺β细胞功能的有效方法。胰腺激素胰岛素已被使用数十年来维持1型糖尿病患者的葡萄糖稳态，而从干细胞中产生的β细胞的产生仍然是许多研究人员作为未来治疗的许多研究人员的重点。在小鼠中的研究表明，Neurog3是一种治疗性纤维素，可从ES细胞产生胰腺内分泌细胞或通过成人细胞类型的新生成。该提案将明确确定beta细胞和肠肠肠肠内分泌的发展是否或不是人类所需的neurog3。无论哪种情况，这些研究都将通过确定Neurog3的相关性并确定与人类β细胞开发有关的补偿性替代途径来确定糖尿病研究界的广泛影响。