Intersection of signaling pathways and transcription factors regulating islet development

调节胰岛发育的信号通路和转录因子的交叉点

• 批准号: 9922264
• 负责人: PAUL J GADUE
• 金额: $ 54.14万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9922264
• 关键词: Address; Affect; Alleles; B-Lymphocytes; Beta Cell; Binding; Biological Models; Biology; Blood Glucose; Cell Line; Cell Therapy; Cell secretion; Cells; Cellular biology; ChIP-seq; Code; D Cells; Data; Development; Diabetes Mellitus; Disease; Disease Management; Disease model; Dominant-Negative Mutation; Endocrine; Enhancers; Failure; Family member; GATA4 gene; GATA6 transcription factor; Gene Dosage; Gene Expression Regulation; Genes; Genetic; Genetic Diseases; Goals; Healthcare; Human; Impairment; In Vitro; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Islets of Langerhans; Knowledge; Light; Mediator of activation protein; Minor; Modeling; Mus; Mutation; Non-Insulin-Dependent Diabetes Mellitus; Pancreas; Pancreatic Diseases; Pathway interactions; Patients; Penetrance; Phenocopy; Phenotype; Protocols documentation; RXR; Regulation; Resources; Rodent; Rodent Model; Role; Signal Pathway; Signal Transduction; Single Nucleotide Polymorphism; Source; System; Tretinoin; United States; base; cost; diabetes mellitus therapy; embryonic stem cell; endocrine pancreas development; human disease; human embryonic stem cell; human pluripotent stem cell; human stem cells; in vivo; induced pluripotent stem cell; insight; insulin secretion; model design; mouse model; mutant; neonatal diabetes mellitus; pancreas development; progenitor; small molecule; standard of care; statistics; stem cell model; stem cells; synergism; transcription factor; transcriptome sequencing

Diabetes is an immense healthcare burden, with great costs for management of this disease and its associated complications. Human pluripotent stem cells (PSC)s, including embryonic stem (ES) cells and induced pluripotent stem (iPS) cells offer great potential for the study and treatment of human disease. Rodent models have also been critical in understanding pancreas development and generating models of diabetes. Here, we propose to use both complementary systems to study human pancreatic development and function, with a focus on the beta cell, which regulates blood glucose via insulin secretion. While rodent studies have produced tremendous advancements, human and mice are not the same and it is important to define differences in pancreas biology as these could impact the treatment of diabetes. For example, heterozygous mutations in the transcription factors GATA6 or GATA4 have been shown to cause pancreas agenesis in humans, with mutations in GATA6 being the most common cause of this genetic disease (~50%). In contrast to the human phenotype, mice heterozygous for either GATA6 or GATA4 have no pancreatic phenotype. Compound mutations of GATA4 and GATA6 do mimic the human disease suggesting an overall conservation in GATA family members in pancreas development but with humans being much more sensitive to dosage of these genes. Retinoic Acid (RA) signaling is a critical mediator of pancreas development both in rodent models and is a required signaling input in directed differentiation protocols generating beta cells from human PSCs. RA is also a known regulator of GATA6 and GATA4 expression. We will examine the interplay between RA and GATA4/6 in both the mouse and human PSC model systems. Specifically, will use mutants in GATA family members as well as dominant negative RXR constructs in both systems, assessing conservation and differences in their impact on pancreas specification and endocrine cell development. Lastly, we will study a single nucleotide polymorphism in the 5’ GATA6 enhancer, which in preliminary data we demonstrate regulates GATA6 expression and pancreas development in the human PSC system. These studies will help define the developmental and functional differences between human and mice beta cells that have the potential to directly impact the treatment of not only rare genetic forms of diabetes but also reveal unique human beta cell biology with implications in the management of both type 1 and type 2 diabetes.

糖尿病是一个巨大的医疗保健负担，管理这种疾病及其相关并发症的成本很高。人类多能干细胞（PSC）包括胚胎干细胞（ES）和诱导多能干细胞（iPS），为人类疾病的研究和治疗提供了巨大的潜力。啮齿动物模型在了解胰腺发育和生成糖尿病模型方面也至关重要。在这里，我们建议使用这两个互补系统来研究人类胰腺的发育和功能，重点是β细胞，它通过胰岛素分泌来调节血糖。虽然啮齿动物研究取得了巨大的进步，但人类和小鼠并不相同，重要的是要确定胰腺生物学的差异，因为这些差异可能会影响糖尿病的治疗。例如，转录因子GATA 6或GATA 4中的杂合突变已被证明会导致人类胰腺发育不全，其中GATA 6中的突变是这种遗传疾病的最常见原因（约50%）。与人类表型相反，GATA 6或GATA 4杂合子小鼠没有胰腺表型。GATA 4和GATA 6的复合突变确实模拟了人类疾病，表明胰腺发育中加塔家族成员的总体保守性，但人类对这些基因的剂量更为敏感。视黄酸（RA）信号传导是啮齿动物模型中胰腺发育的关键介质，并且是从人PSC产生β细胞的定向分化方案中所需的信号传导输入。RA也是已知的GATA 6和GATA 4表达的调节剂。我们将研究RA和GATA 4/6在小鼠和人PSC模型系统中的相互作用。具体而言，将使用突变体在加塔家庭成员以及显性负RXR结构在这两个系统中，评估其对胰腺规格和内分泌细胞发育的影响的保守性和差异。最后，我们将研究5'GATA 6增强子中的单核苷酸多态性，在初步数据中，我们证明该多态性调节人PSC系统中的GATA 6表达和胰腺发育。这些研究将有助于确定人类和小鼠β细胞之间的发育和功能差异，这些差异不仅有可能直接影响罕见遗传形式的糖尿病的治疗，而且还揭示了独特的人类β细胞生物学，对1型和2型糖尿病的管理都有影响。