The Pdx1-recruited Swi/Snf chromatin remodeling complex regulates endocrine cell expansion and differentiation in vivo

Pdx1 招募的 Swi/Snf 染色质重塑复合物调节体内内分泌细胞的扩增和分化

基本信息

项目摘要

PROJECT SUMMARY/ABSTRACT Pancreatic β-cells within the islets of Langerhans are required for glucose-stimulated insulin secretion and glucose homeostasis. Dysfunctional β-cell activity and identity results in diabetes mellitus (DM), a growing disease affecting millions of Americans, thus creating an enormous fiscal and health burden. Strategies to improve outcomes for the mounting number of diabetic patients and the possibility to replace destroyed and dysfunctional β-cell mass requires a deep understanding of the complex programs that coordinate proper islet formation. Developing upon existing knowledge of how islet enriched transcription factors (TFs) coordinate signals that direct islet cell development will allow us to understand how such programs can be targeted for β- cell replacement therapies. Pdx1, one of the most important TFs in the pancreas and developing islet, has been shown to recruit a diverse set of coregulators which could potentially modulate its activity. This proposal is focused around how the Pdx1 recruited ATP-dependent Swi/Snf chromatin remodeling complex modulates Pdx1 transcriptional activity in the developing islet. Previous work by the PI’s group revealed a critical role for Pdx1:Swi/Snf function in the developing pancreas and the mature β-cell. Preliminary studies thus far have shown that conditional removal of one of the core Swi/Snf ATPase subunits (through Cre-LoxP technology) from developing endocrine progenitor cells results in glucose intolerance, ad libitum fed hyperglycemia and reduced serum insulin levels in mutant animals postnatally. This proposal will test the hypothesis that Pdx1-recruited Swi/Snf chromatin remodeling activities control chromatin accessibility and gene expression programs essential for endocrine progenitor cell expansion and postnatal islet cell function. In Aim 1, the PI will use this novel transgenic animal model to determine islet cell function, expression of islet cell maturation markers, hormone secretion and hormone cell mass in adult pancreata. In Aim 2, embryonic tissues from mutant animals will be used to determine changes in endocrine progenitor cell mass and hormone cell proliferation/apoptosis. The mechanistic actions of Swi/Snf on directing chromatin accessibility and gene expression programs in endocrine progenitors will be determined by ATAC-Seq, RNA-Seq and ChIP-qPCR analyses on flow-sorted Swi/Snf-deficient endocrine precursors. My extensive experience studying transcription factor coregulatory complexes and our available in vivo reagents make my lab uniquely suited to accomplish these Aims. Mechanisms defined by these studies will uncover key processes and unique gene expression signatures dependent on chromatin remodeling during endocrine development, which will provide instrumental knowledge to researchers developing new molecular targets and cell-based therapies to combat diabetes. Importantly, this proposal has also been designed to generate data and resources that the applicant will utilize in an R01 application defining how dysregulated enhancer function from Type 2 diabetic (T2D) islet donors are influenced by altered Pdx1:Swi/Snf actions.
项目摘要/摘要 胰岛内的胰腺β细胞是葡萄糖刺激的胰岛素分泌所必需的 和葡萄糖稳态。功能失调的β细胞活性和身份导致糖尿病(DM),这是一种日益严重的糖尿病。 疾病影响了数百万美国人,从而造成了巨大的财政和健康负担。的战略 改善越来越多的糖尿病患者的预后,并有可能取代被破坏的, 功能失调的β细胞群需要深入了解协调适当胰岛的复杂程序, 阵发展现有的知识,如何胰岛富集转录因子(TF)协调 指导胰岛细胞发育的信号将使我们了解这些程序如何针对β- 细胞替代疗法 Pdx 1是胰腺和发育中胰岛中最重要的TF之一,已被证明可以招募一种新的转录因子。 可能潜在地调节其活性的多种辅助调节因子。该提案的重点是如何 Pdx 1募集ATP依赖性Swi/Snf染色质重塑复合物调节Pdx 1转录活性 在发展中的小岛。PI小组先前的工作揭示了Pdx 1:Swi/Snf功能在细胞凋亡中的关键作用。 发育中的胰腺和成熟的β细胞。迄今为止的初步研究表明, 一个核心Swi/Snf ATP酶亚基(通过Cre-LoxP技术)来自发育中的内分泌祖细胞 细胞导致葡萄糖耐受不良,随意喂养高血糖症和降低血清胰岛素水平的突变 动物产后该提议将检验Pdx 1募集Swi/Snf染色质重塑的假设, 活性控制内分泌祖细胞所必需的染色质可及性和基因表达程序 扩张和出生后胰岛细胞功能。在目标1中,PI将使用这种新型转基因动物模型, 测定胰岛细胞功能、胰岛细胞成熟标志物表达、激素分泌和激素细胞 成人胰腺肿块。在目标2中,来自突变动物的胚胎组织将用于确定 内分泌祖细胞质量和激素细胞增殖/凋亡。Swi/Snf的作用机制 指导内分泌祖细胞中的染色质可及性和基因表达程序将由以下决定 流式分选Swi/Snf缺陷型内分泌前体的ATAC-Seq、RNA-Seq和ChIP-qPCR分析。 我在研究转录因子共调节复合物方面的丰富经验以及我们在体内的可用性 试剂使我的实验室特别适合实现这些目标。这些研究确定的机制将 揭示关键过程和独特的基因表达签名依赖于染色质重塑过程中 内分泌发展,这将提供工具知识的研究人员开发新的分子 靶点和细胞疗法来对抗糖尿病。重要的是,该提案还旨在 生成申请人将在R 01申请中使用的数据和资源,定义如何失调 来自2型糖尿病(T2 D)胰岛供体的增强子功能受到改变的Pdx 1:Swi/Snf作用的影响。

项目成果

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Jason M Spaeth其他文献

Jason M Spaeth的其他文献

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{{ truncateString('Jason M Spaeth', 18)}}的其他基金

The role of Pdx1-recruited Chd4:NuRD complex in controlling mature #-cell function
Pdx1招募的Chd4:NuRD复合物在控制成熟中的作用
  • 批准号:
    10634693
  • 财政年份:
    2022
  • 资助金额:
    $ 11.89万
  • 项目类别:
Islet transcription factor activation: FoxPs are required for postnatal endocrine cell proliferation while Pdx1 recruited chromatin remodeling enzymes impact pancreas size
胰岛转录因子激活:FoxPs 是出生后内分泌细胞增殖所必需的,而 Pdx1 招募的染色质重塑酶会影响胰腺大小
  • 批准号:
    8907572
  • 财政年份:
    2015
  • 资助金额:
    $ 11.89万
  • 项目类别:
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