Human Endocrine Cell Development

人类内分泌细胞发育

基本信息

批准号：
8456068
负责人：
NOAH Freeman SHROYER
金额：
$ 43.91万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
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/8456068
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

项目摘要

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。