The role of GATA6 and GATA4 in pancreatic beta cell function and development

GATA6 和 GATA4 在胰腺 β 细胞功能和发育中的作用

• 批准号: 9395270
• 负责人: Karla Fitzgerald Leavens
• 金额: $ 6.74万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9395270
• 关键词: Affect; Apoptosis; Beta Cell; Biological Models; Calcium Signaling; Cell Differentiation process; Cell Line; Cell model; Cell physiology; Cells; Complex; Data; Defect; Development; Diabetes Mellitus; Disease; Endoderm; Excision; Family; Family member; Flow Cytometry; Functional disorder; GATA4 gene; GATA6 transcription factor; Genes; Genetic Engineering; Genomics; Glucose; Goals; Heterogeneity; Human; Human Engineering; In Vitro; Individual; Insulin; Investigation; Knock-out; Knowledge; LoxP-flanked allele; Measures; Modeling; Modification; Molecular; Mus; Mutation; Pancreas; Pathway interactions; Patients; Penetrance; Phenotype; Physiological; Play; Protein Isoforms; Proteins; Research; Role; Severities; Signal Induction; Signal Transduction; Small Interfering RNA; Stimulus; Structure of beta Cell of islet; System; Techniques; Tretinoin; Western Blotting; Zinc Fingers; blood glucose regulation; experimental study; glucose uptake; human pluripotent stem cell; in vivo; insulin secretion; knock-down; member; overexpression; pancreas development; response; technique development; therapeutic target; therapy development; transcription factor

Project Summary/Abstract GATA6 and GATA4 are two members of the GATA family of zinc finger transcription factors and have been shown to be important for pancreatic development. Heterozygous mutations in GATA6 and GATA4 account for the majority of cases of patients with pancreatic agenesis. However, GATA6 and GATA4 clearly play a more complex role in pancreatic development and function as these same heterozygous mutations can result in diabetes of varying onset and severity in some individuals without pancreatic agenesis, differing even within family members carrying the same mutations. Factors that cause disease due to haploinsufficiency can be particularly important therapeutic targets as small modifications in their action could be the difference between disease and normal function. Therefore, understanding the role of the GATA factors will advance our knowledge of beta cell function. Though these proteins have been studied in mice, they do not recapitulate the human phenotype, making studies in human cells necessary. The development of techniques to differentiate human pluripotent stem cells (hPSCs) into pancreatic cells has allowed for the use of a human system in which to accomplish these investigations. Preliminary data from our group show that GATA6 is necessary for normal endoderm induction during differentiation of hPSCs, and that loss of GATA6 results in defective glucose-stimulated insulin secretion in differentiated pancreatic beta cells. Additionally, preliminary studies show that GATA6 and GATA4 expression can be induced by retinoic acid, one of the inductive signals involved in the differentiation of pancreatic cells, suggesting interaction between the two pathways. The research described in this proposal will use two human models of pancreatic beta cells, differentiated hPSCs and a human beta cell line, to characterize the role of GATA6 and GATA4 in glucose-stimulated insulin secretion and to define the interaction between the GATA factors and retinoic acid signaling. By selectively knocking down and overexpressing the GATA factors in mature pancreatic beta cells, we will be able to determine their effect on insulin secretion independent of their effect on development. Utilizing this same technique in both differentiating and mature pancreatic beta cells while modifying retinoic acid signaling, we will be able to characterize the interaction between GATA4/6 and retinoic acid. These studies will help advance our knowledge of pancreatic beta cell function and development and therefore allow for a better understanding of the pathophysiology underlying diseases resulting from their dysfunction, including diabetes.

项目总结/摘要 GATA 6和GATA 4是锌指转录因子加塔家族的两个成员， 对胰腺发育很重要。GATA 6杂合突变， GATA 4占胰腺发育不全患者的大多数病例。然而，GATA 6和 GATA 4在胰腺发育和功能中显然起着更复杂的作用， 杂合突变可导致糖尿病的不同发病和严重程度，在一些个体中， 胰腺发育不全，甚至在携带相同突变的家庭成员中也不同。的因素 因单倍性不足引起的疾病可以作为特别重要的小治疗靶点 其作用的改变可能是疾病和正常功能之间的区别。因此，我们认为， 理解加塔因子的作用将促进我们对β细胞功能的认识。虽然 这些蛋白质已经在小鼠中进行了研究，它们不重现人类表型， 人类细胞的研究是必要的。人类多能分化细胞技术的发展 干细胞（hPSC）转化为胰腺细胞的方法已经允许使用人类系统， 完成这些调查。我们小组的初步数据表明，GATA 6是必要的， 在hPSC分化过程中正常的内胚层诱导，并且GATA 6的缺失导致缺陷的内胚层诱导。 在分化的胰腺β细胞中葡萄糖刺激的胰岛素分泌。此外，初步 研究表明，GATA 6和GATA 4的表达可以被视黄酸诱导，视黄酸是诱导GATA 6和GATA 4表达的诱导剂之一。 参与胰腺细胞分化的信号，表明两者之间的相互作用 途径。该提案中描述的研究将使用两种胰腺β细胞的人类模型， 本发明涉及一种用于检测分化的hPSC和人β细胞系中GATA 6和GATA 4的表达的方法，以表征GATA 6和GATA 4在分化的hPSC和人β细胞系中的作用。 葡萄糖刺激的胰岛素分泌，并确定加塔因子之间的相互作用， 视黄酸信号传导。通过选择性地敲低和过表达加塔因子， 胰腺β细胞，我们将能够确定它们对胰岛素分泌的影响，而不依赖于它们对胰岛素分泌的影响。 对发展的影响。在分化和成熟的胰腺β细胞中使用相同的技术 细胞，同时修改视黄酸信号，我们将能够表征之间的相互作用 GATA 4/6和视黄酸。这些研究将有助于我们进一步了解胰腺β细胞 功能和发展，因此可以更好地了解病理生理学 由其功能障碍引起的潜在疾病，包括糖尿病。