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。

基本信息

批准号：
8846104
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
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8846104
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 Genetic screening method Glucagon Glucose Health 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 skills tool transcription factor type I and type II diabetes

项目摘要

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）：Langerhans胰岛内的胰腺β细胞是葡萄糖刺激的胰岛素分泌和葡萄糖稳态所必需的。 Beta细胞活动或生存的功能障碍导致糖尿病梅洛蒂斯（Mellitus），这种疾病目前会影响美国人的数百万美元，预计将来的数量将大大增加，从而造成了巨大的经济和健康负担。改善糖尿病患者数量不断增长的结果的未来策略需要了解指定和区分功能性β细胞与祖细胞的复杂发育计划。利用对β细胞生物学中心的信号传导和转录事件的现有知识以及未来的理解将使您能够产生治疗性替代β细胞进行移植。这些目标的核心是假设，即由Lim域转录因子介导的基因调节所必需的转录辅助因子，以在发育和成人期间产生功能性β细胞。我的初步数据定义了胰腺中LDB1的表达模式以及LDB1在发育内分泌细胞中的作用。有条件地删除的小鼠证明，LDB1发育功能部分与胰腺中唯一良好研究的LIM因子（ISL1）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。 AIM 1将利用无偏基因组微阵列和芯片序列方法比较胰腺发育过程中由LDB1和ISL1调节的基因和途径。 AIM 2将使用创新的免疫沉淀程序隔离并比较胰腺内分泌细胞的LDB1-和ISL1相互作用的共同调节剂，因为我假设LDB1和ISL1和ISL1募集了胰腺相互作用因子的独特子集。在AIM 3中，我将测试新的AIM 2候选蛋白在利用体内敲除和细胞系敲除模型的LDB1或ISL1介导的转录控制中的潜在作用。 LDB1：ISL1共享目标基因在内，包括MAFA和ARX或AIM 1中鉴定的目标基因将通过新的交互者进行调节测试。有了这一K01职业发展奖，我将测试我的总体假设，即LDB1控制着一系列胰岛靶基因，其中包括ISL1-和（可能）LMO介导的复合物，部分原因是与不同的结合核心基因相互作用。我当前的环境非常适合我成功启动拟议的研究和进一步培训。我将开发工具和技能，以回答这些目标提出的许多问题并产生有趣的数据，从而使我能够作为独立研究人员过渡到资金。