The Ldb1 coregulator controls LIM target genes in developing and adult islets.

Ldb1 核心调节器控制发育中和成年胰岛中的 LIM 靶基因。

• 批准号: 8710206
• 负责人: Chad S Hunter
• 金额: $ 15.2万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8710206
• 关键词: Adult; Affect; Alpha Cell; American; Automobile Driving; Award; Beta Cell; Binding; Binding Proteins; Biochemical; Birth; Cell Count; Cell Line; Cell Maturation; Cell Transplantation; Cells; Cellular biology; Collaborations; Complex; DNA Binding; Data; Data Set; Development; Developmental Process; Diabetes Mellitus; Disease; Ductal Epithelial Cell; Endocrine; Environment; Erythroid Cells; Event; Excision; Fibrinogen; Fluorescence-Activated Cell Sorting; Functional disorder; Funding; Future; Gene Expression; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genetic Programming; Glucagon; Glucose; Health Care Costs; High-Throughput Nucleotide Sequencing; Immunoprecipitation; In Vitro; Insulin; Islet Cell; Islets of Langerhans; Knock-out; Knockout Mice; Knowledge; LIM Domain; Laboratories; Link; Mass Spectrum Analysis; Measures; Mediating; Modeling; Mus; Occupations; Organogenesis; Outcome; Pancreas; Pathway interactions; Patients; Pattern; Phenotype; Population; Procedures; Processed Genes; Property; Proteins; Protocols documentation; Quality of life; RNA; Recruitment Activity; Regulation; Relative (related person); Research Personnel; Role; SS DNA BP; Signal Transduction; Somatostatin; Specific qualifier value; Specificity; Spinal Cord; Structure of beta Cell of islet; Testing; Therapeutic; Tissues; Training; Transcription factor genes; Transcriptional Regulation; Transgenes; Transplantation; United States National Institutes of Health; base; blood glucose regulation; career; chromatin immunoprecipitation; cofactor; combat; crosslink; developmental genetics; diabetes mellitus therapy; diabetic patient; economic cost; genome-wide; health economics; homeodomain; improved; in vivo; innovation; insight; insulin secretion; interest; islet; novel; pancreas development; postnatal; progenitor; programs; promoter; public health relevance; skills; tool; transcription factor; type I and type II diabetes

DESCRIPTION (provided by applicant): Pancreatic beta cells within the Islets of Langerhans are required for glucose-stimulated insulin secretion and glucose homeostasis. Dysfunction in beta cell activity or survival results in diabetes mellitus, a disease currently affecting millionsof Americans with numbers expected to greatly increase in the future, thus creating an enormous economic and health burden. A future strategy to improve outcomes for the growing numbers of diabetic patients requires understanding the complex developmental programs that specify and differentiate functional beta cells from progenitors. Exploiting existing knowledge and future understanding of the signaling and transcriptional events central to beta-cell biology will allow u to produce therapeutic replacement beta cells for transplantation. Central to these Aims is the hypothesis that the transcriptional cofactor, Ldb1, is required for gene regulation mediated by LIM-domain transcription factors to produce functional beta cells during development and in adults. My preliminary data defines the expression pattern of Ldb1 in the pancreas as well as the role of Ldb1 in developing endocrine cells. Conditionally deleted mice demonstrated that Ldb1 developmental function partially overlaps with the only well-studied LIM factor in the pancreas, Islet-1 (Isl1). These Aims will further define the Isl1-dependent and -independent genetic events controlled by Ldb1. Aim 1 will utilize unbiased genome-wide microarray and ChIP-Seq approaches to compare the genes and pathways regulated by Ldb1 and Isl1 during pancreas development. Aim 2 will isolate and compare Ldb1- and Isl1-interacting co-regulators from pancreatic endocrine cells using an innovative immunoprecipitation procedure, as I hypothesize that Ldb1 and Isl1 recruit unique subsets of interacting factors in the pancreas. In Aim 3, I will test new Aim 2 candidate binding proteins for their potential roles in Ldb1- or Isl1-mediated transcriptional control utilizing in vivo knockout and cell line knockdown models. Ldb1:Isl1 shared target genes including MafA and Arx or those identified in Aim 1 will be tested for regulation by new interactors. With this K01 Career Developmental Award, I will test my overall hypothesis that Ldb1 controls an array of islet target genes that include Isl1- and (potentially) LMO-mediated complexes in part through interactions with distinct binding coregulators. My current environment is uniquely suited for me to successfully initiate the proposed studies and further training. I will develop the tools and skills to answer many of the questions raised by these Aims and generate interesting data allowing me to transition into funding as an independent investigator.

描述（由申请人提供）：朗格汉斯胰岛内的胰腺β细胞是葡萄糖刺激胰岛素分泌和葡萄糖稳态所必需的。β细胞活性或存活功能障碍导致糖尿病，这种疾病目前影响着数百万美国人，预计未来人数将大幅增加，从而造成巨大的经济和健康负担。为了改善越来越多的糖尿病患者的预后，未来的策略需要了解复杂的发育程序，这些程序指定并区分功能性β细胞和祖细胞。利用现有的知识和未来对β细胞生物学中心的信号和转录事件的理解，将使我们能够生产用于移植的治疗性替代β细胞。这些目标的核心假设是转录辅助因子Ldb1在发育和成人过程中由lim结构域转录因子介导的基因调控中是必需的，以产生功能性β细胞。我的初步数据定义了Ldb1在胰腺中的表达模式以及Ldb1在发育内分泌细胞中的作用。条件缺失小鼠表明，Ldb1发育功能与胰腺中唯一被充分研究的LIM因子Islet-1 （Isl1）部分重叠。这些目标将进一步定义由Ldb1控制的isl1依赖性和非依赖性遗传事件。目的1将利用无偏倚的全基因组微阵列和ChIP-Seq方法来比较Ldb1和Isl1在胰腺发育过程中调控的基因和途径。Aim 2将使用创新的免疫沉淀程序从胰腺内分泌细胞中分离和比较Ldb1和Isl1相互作用的共同调节因子，因为我假设Ldb1和Isl1在胰腺中招募了独特的相互作用因子亚群。在Aim 3中，我将利用体内敲除和细胞系敲除模型测试新的Aim 2候选结合蛋白在Ldb1或isl1介导的转录控制中的潜在作用。Ldb1:Isl1共有的靶基因包括MafA和Arx或Aim 1中发现的靶基因，将测试新的相互作用物的调控作用。有了这个K01职业发展奖，我将测试我的总体假设，即Ldb1控制一系列胰岛靶基因，其中包括Isl1和（潜在的）lmo介导的复合物，部分通过与不同的结合共调节因子相互作用。我目前的环境非常适合我成功地开始计划的学习和进一步的培训。我将开发工具和技能来回答这些目标提出的许多问题，并生成有趣的数据，使我能够以独立调查员的身份过渡到资助。