Understanding human pancreas development for diabetes cell-replacement therapy

了解糖尿病细胞替代疗法的人类胰腺发育

基本信息

批准号：
10215481
负责人：
Danwei Huangfu
金额：
$ 57.01万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
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：
2012-07-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10215481
关键词：
ATAC-seq Affect B cell differentiation Beta Cell Binding Biochemical Cell Count Cell Line Cell physiology Cells ChIP-seq Chromatin Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Complex Cues Data Defect Development Diabetes Mellitus Disease Disease model Dose Etiology GATA4 gene Generations Genes Genetic Genomic approach Genomics Genotype Grant Health Human Human Development Hyperinsulinism Hypoglycemia Impairment Insulin Insulin-Dependent Diabetes Mellitus Investigation Lead Mediating Missense Mutation Modeling Molecular Mus Mutation Neonatal Non-Insulin-Dependent Diabetes Mellitus Pancreas Pancreatic Diseases Patients Phenotype Point Mutation Publications Publishing Resolution Signal Transduction Structure of beta Cell of islet System Time Transcriptional Regulation Variant Work base beta cell replacement cell replacement therapy diabetes mellitus therapy differentiation protocol disease phenotype dosage genetic approach genetic manipulation genome editing human disease human pluripotent stem cell improved islet stem cells maturity onset diabetes of the young mouse model mutant neonatal diabetes mellitus novel pancreas development progenitor protein function single cell technology stem cell differentiation stem cell model tool transcription activator-like effector nucleases transcription factor transcriptomics unpublished works

项目摘要

Delineating key factors that regulate early pancreas development is crucial to our long-term pursuit of understanding disease mechanisms and developing human pluripotent stem cell (hPSC) based β-cell replacement therapies for diabetes. However, a complete landscape of signaling cues and transcription factors required for pancreas specification remains unclear. Using hPSC directed differentiation and CRISPR/Cas-mediated genome editing, recently published work from the Huangfu lab has identified critical new requirements for RFX6, GATA6 and GATA4 (genes known to be associated with neonatal and adult-onset diabetes) during human pancreatic differentiation. In unpublished work, we have further uncovered a novel, dose-dependent requirement for FOXA2, a gene associated with diabetes and hyperinsulinism, in pancreatic differentiation. Here in this collaboration between the Huangfu, Leslie and Pe’er labs, we will undertake complementary genetic, genomic and computational approaches and utilize hPSC differentiation to dissect human pancreatic development. We will utilize genetic approach to create precise hPSC disease models and interrogate complex genetic interactions underlying disease phenotypes, and employ genomic approaches including ChIP-seq and ATAC-seq analyses and cutting-edge single-cell transcriptomics to understand regulators of human pancreatic development and β cell function. Our findings will enhance the understanding of human pancreatic development and disease, and facilitate the development of improved hPSC directed differentiation protocols for the generation of functional β cells for disease study and treatment.

描绘早期调节胰腺发育的关键因素对于我们长期追求了解疾病机制和开发基于人类多能干细胞 (hPSC) 的糖尿病 β 细胞替代疗法至关重要。皇甫实验室最近发表的研究成果利用 hPSC 定向分化和 CRISPR/Cas 介导的基因组编辑，确定了 RFX6、GATA6 和 GATA4（已知与胰腺相关的基因）的关键新要求。在未发表的工作中，我们进一步发现了 FOXA2（一种与糖尿病和高胰岛素血症相关的基因）在胰腺分化过程中的一种新的剂量依赖性需求。 Leslie 和 Pe'er 实验室，我们将采用互补的遗传、基因组和计算方法，并利用 hPSC 分化来剖析人类胰腺发育。我们将利用遗传方法创建精确的 hPSC 疾病模型并探究遗传复杂的相互作用。潜在的疾病表型，并采用包括 ChIP-seq 和 ATAC-seq 分析以及尖端单细胞转录组学在内的基因组方法来了解人类胰腺发育和 β 细胞功能的调节因子，我们的研究结果将增强对人类胰腺发育和疾病的理解，并促进改进 hPSC 定向分化方案的开发，以生成用于疾病研究和治疗的功能性 β 细胞。