CHD4-MEDIATED TRANSCRIPTIONAL REGULATION OF PANCREATIC BETA CELL DEVELOPMENT AND FUNCTION

CHD4 介导的胰腺 β 细胞发育和功能的转录调控

• 批准号: 10656226
• 负责人: Dylan Sarbaugh
• 金额: $ 3.93万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656226
• 关键词: Adult; Affect; Age; Beta Cell; Binding; Biological Assay; Blood Glucose; CHD4 gene; Cadaver; Cell Line; Cell physiology; Cells; ChIP-seq; Chromatin; Co-Immunoprecipitations; Complex; Data; Deacetylase; Defect; Development; Diabetes Mellitus; Donor person; Enterobacteria phage P1 Cre recombinase; Epidemic; Etiology; Functional disorder; Genetic Transcription; Genomics; Glucose; Glucose Intolerance; HDAC1 gene; Hormones; Human; Hyperglycemia; In Vitro; Insulin; Islet Cell; Islets of Langerhans Transplantation; Knowledge; Mass Spectrum Analysis; Mediating; Metabolic; Methods; Molecular; Motor; Mus; Mutant Strains Mice; Non-Insulin-Dependent Diabetes Mellitus; Nucleosomes; Occupations; Pancreas; Pancreas Transplantation; Patients; Play; Population; Proteins; Research Personnel; Role; Severities; Stains; Structure of beta Cell of islet; Testing; Time; Transcriptional Regulation; cell replacement therapy; cofactor; combat; conditional knockout; diabetic patient; differentiation protocol; directed differentiation; endocrine pancreas development; experimental study; gene repression; genetic corepressor; global health; histone modification; human pluripotent stem cell; in vivo; insulin secretion; islet; knock-down; novel; programs; recruit; stem cell differentiation; success; transcription factor; transcriptome sequencing; type I and type II diabetes

PROJECT SUMMARY / ABSTRACT Diabetes is a global epidemic that is expected to become even more severe. Although there are treatment options, such as exogenous insulin, one of the most promising long-term treatments is transplanting donor islets into diabetic patients. While this treatment can allow patients to maintain normal blood glucose levels for extended periods of time, there is an extreme lack in the amount of viable donor islets for transplantation. To combat the lack of suitable donor islets, researchers have begun to differentiate human pluripotent stem cells (hPSCs) into β-like cells that respond to glucose by secreting insulin, with the hopes of creating a limitless supply of transplantable pancreatic β cells. This approach has been somewhat successful; however, the current differentiation protocols lack the ability to make pure populations of β cells and often result in immature β cells and polyhormonal cell populations. More complete knowledge of the transcriptional networks and associated cofactors required in the differentiation and function of pancreatic β cells is needed to supplement the current hPSC differentiation protocols and provide the information required for better diabetes treatment options. One such cofactor is the chromodomain helicase DNA-binding protein 4 (CHD4). CHD4 is the motor protein of the nucleosome remodeling and deacetylase (NuRD) complex and, along with histone deacetylase 1 and 2 (HDAC1 and 2), CHD4 creates condensed chromatin states, thereby repressing genes. Preliminary data has shown that CHD4 interacts with essential transcription factors in pancreatic β cells. Furthermore, my preliminary data shows that the loss of CHD4 in the β cells of mice causes hyperglycemia along with glucose intolerance. This suggests that CHD4 is a necessary transcriptional cofactor in the function of β cells. A better understanding of the role CHD4 plays in the development and function of pancreatic β cells could provide additional information to facilitate the directed differentiation of β cells from hPSCs in vitro. In this proposal, using both in vivo and in vitro experiments, I will determine the role and mechanism of CHD4 in the development and function of pancreatic β cells.

项目摘要 /摘要 糖尿病是一种全球流行病，预计将变得更加严重。虽然有治疗 诸如外源胰岛素之类的选择，最有希望的长期治疗之一是移植供体胰岛 进入糖尿病患者。尽管这种治疗可以使患者保持正常的血糖水平 长时间的时间，可行的供体胰岛的移植数量极为缺乏。到 抗击缺乏合适的供体胰岛，研究人员已经开始区分人类多能干细胞 （HPSC）进入β样细胞，通过分泌胰岛素对葡萄糖反应，希望产生有限的供应 可移植的胰腺β细胞。这种方法有些成功。但是，电流 分化方案缺乏使纯β细胞群体的能力，并且通常导致未成熟的β细胞 和多气压细胞群。对转录网络和相关的更完整的了解 需要在胰腺β细胞的分化和功能中需要的辅助因子来补充电流 HPSC分化方案，并提供更好的糖尿病治疗选择所需的信息。一 这种辅助因子是染色体构酶DNA结合蛋白4（CHD4）。 CHD4是运动蛋白 核小体重塑和脱乙酰基酶（NURD）复合物以及组蛋白脱乙酰基酶1和2（HDAC1 和2），CHD4产生凝结的染色质状态，从而反映基因。初步数据表明 CHD4与胰腺β细胞中基本转录因子相互作用。此外，我的初步数据显示 小鼠β细胞中CHD4的丧失会引起高血糖症和葡萄糖肠道。这暗示着 该CHD4是β细胞功能中必要的转录辅助因子。更好地理解角色 CHD4在胰腺β细胞的开发和功能中发挥作用，可以提供其他信息以促进 在体外，β细胞与HPSC的定向分化。在此提案中，使用体内和体外都 实验，我将确定CHD4在开发和功能中的作用和机制 胰腺β细胞。