Regulation of kidney-Specific Gene Expression

肾脏特异性基因表达的调节

基本信息

批准号：
7988995
负责人：
Peter Igarashi
金额：
$ 9.26万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-12-07 至 2011-12-06
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7988995
关键词：
Animal Model Autosomal Recessive Polycystic Kidney Binding Sites Biochemical Biological Assay C-terminal Cell Differentiation process Child Chromatin Structure Cyst Cystic Kidney Diseases Cystic kidney DNA Binding Development Down-Regulation Duct (organ) structure Epithelial Exhibits Family Gene Expression Gene Targeting Genes Genetic Genetic Recombination Genetic Transcription Genitourinary system Goals Histone Acetylation Histones Human Infant Kidney Kidney Failure Knock-in Mouse LacZ Genes Ligase Link Liver Location Lysine Maps Measures Mediating Microarray Analysis Modification Molecular Mus Mutant Strains Mice Mutate Mutation Non-Insulin-Dependent Diabetes Mellitus Organ Organogenesis PKHD1 gene Pancreas Pathogenesis Pattern Phenotype Process Proteins Regulation Regulatory Element Reporter Genes Research Personnel Role Site Testing Tissue-Specific Gene Expression Tissues Transcriptional Activation Transcriptional Activation Domain Transgenic Mice Transgenic Organisms Ubiquitin Ubiquitin-Conjugating Enzymes Ubiquitination Yeasts bile duct deletion analysis disease phenotype hepatocyte nuclear factor histone acetyltransferase homeodomain in vivo inhibitor/antagonist insight kidney cell mutant nephrogenesis novel programs promoter transcription factor ubiquitin ligase yeast two hybrid system

项目摘要

The overall goal of this project is to understand the roles of hepatocyte nuclear factor-13 (HNF-1(3)in kidney-specific gene expression, renal cell differentiation, and kidney organogenesis. HNF-1(3belongs to a family of homeodomain-containing transcription factors that regulate tissue-specific gene expression in the kidney, liver, pancreas, and other organs. Humans with mutations of HNF-13 develop maturity-onset diabetes of the young type 5 (MODY5) and congenital cystic abnormalities of the kidney. Transgenic mice expressing mutant HNF-13 under the control of a kidney-specific promoter develop kidney cysts and renal failure, which is similar to the phenotype of humans with MODY5. Similarly, kidney-specific deletion of HNF- 1(3 using Cre/loxP recombination results in renal cyst formation. HNF-1{3mutant mice show decreased expression of Pkhdl, the gene mutated in autosomal recessive polycystic kidney disease (ARPKD), and HNF-13 directly regulates the Pkhdl promoter. These studies identify Pkhdl as a novel gene target of HNF- 13 in the kidney. They establish a previously unrecognized link between two renal cystic diseases, MODY5 and ARPKD, and suggest that the mechanism of cyst formation in humans with MODY5 involves down- regulation of PKHD1 gene expression. To test this hypothesis and to further define the functions of HNF-13 in the kidney, we will complete the following specific aims: 1. Define the roles of coactivators and histone acetylation in the regulation of Pkhdl gene transcription. 2. Determine how the activity of HNF-13 is reciprocally regulated by SUMO ligases and ubiquitin ligases. 3. Determine how HNF-13 regulates the tissue-specific expression of Pkhdl in the kidney and liver. 4. Elucidate the molecular pathogenesis of the kidney and genitourinary tract abnormalities caused by mutations of HNF-13. The proposed studies will utilize genetically-modified mice to define in vivo expression patterns and disease phenotypes and biochemical studies to elucidate molecular mechanisms. Understanding the functions of HNF-13 and the regulation of Pkhdl gene transcription will provide insights into the pathogenesis of congenital kidney abnormalities and ARPKD,which is one of the most common genetic causes of renal failure in infants and children.

该项目的总体目标是了解肝细胞核因子 13 (HNF-1(3)) 在肾脏特异性基因表达、肾细胞分化和肾脏器官发生。 HNF-1(3属于含有同源结构域的转录因子家族，可调节组织特异性基因表达肾脏、肝脏、胰腺和其他器官。携带 HNF-13 突变的人类发育成熟年轻 5 型糖尿病 (MODY5) 和先天性肾脏囊性异常。转基因小鼠在肾脏特异性启动子的控制下表达突变型 HNF-13 会形成肾囊肿和肾失败，这与 MODY5 人类的表型相似。同样，肾脏特异性删除 HNF- 1(3 使用 Cre/loxP 重组导致肾囊肿形成。HNF-1{3突变小鼠表现出肾囊肿减少 Pkhdl 的表达，该基因在常染色体隐性多囊肾病 (ARPKD) 中发生突变，以及 HNF-13 直接调节 Pkhd1 启动子。这些研究将 Pkhdl 确定为 HNF-的新基因靶标 13 于肾。他们在两种肾囊性疾病 MODY5 之间建立了先前未被认识的联系和 ARPKD，并表明携带 MODY5 的人类囊肿形成机制涉及下调 PKHD1 基因表达的调控。检验这一假设并进一步定义 HNF-13 的功能在肾脏方面，我们将完成以下具体目标： 1. 定义共激活子和组蛋白乙酰化在 Pkhdl 基因转录调控中的作用。 2. 确定 SUMO 连接酶和泛素连接酶如何相互调节 HNF-13 的活性。 3.确定HNF-13如何调节肾脏和肝脏中Pkhdl的组织特异性表达。 4. 阐明肾脏及泌尿生殖道异常的分子发病机制 HNF-13 突变。拟议的研究将利用转基因小鼠来定义体内表达模式和疾病表型和生化研究以阐明分子机制。了解 HNF-13 的功能和 Pkhdl 基因转录的调控将提供对先天性肾脏异常和 ARPKD 的发病机制，这是最常见的遗传性疾病之一婴儿和儿童肾衰竭的原因。