FUNCTION AND REGULATION OF CD-RAP

CD-RAP的功能和调节

基本信息

批准号：
7940618
负责人：
LINDA J SANDELL
金额：
$ 10.7万
依托单位：
WASHINGTON UNIVERSITY
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：
2009-09-28 至 2010-09-27
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7940618
关键词：
Actins Adenoviruses Adipose tissue Adult Affect American Animal Model Apoptosis Area Awareness Binding Biological Biological Models Biology Blood Vessels Bone Development Bone Marrow Boxing Calcified Candidate Disease Gene Cartilage Cartilage injury Cattle Cell Culture Techniques Cell Cycle Cell Cycle Progression Cell Differentiation process Cells Characteristics Chondrocytes Chondrogenesis Collaborations Collagen Collagen Type II Collagen Type X Data Degenerative Disorder Degenerative polyarthritis Development Disease Down-Regulation Dwarfism EGF gene EWS/FLI 1 Type 1 antisense oligonucleotide Enzymes Epigenetic Process Epiphysial cartilage Event Excision Extracellular Matrix Fibroblast Growth Factor Funding Gene Expression Gene Expression Regulation Genes Genetic Transcription Glyceraldehyde-3-Phosphate Dehydrogenases Goals Grant Growth Factor HMGA Proteins HMGA1 gene HMGA2 gene Human Hypertrophy In Vitro Intervention Knee Knock-out Knowledge Mesenchymal Differentiation Messenger RNA Methods Microarray Analysis Mitogen-Activated Protein Kinases Modeling Mus Oligonucleotide Microarrays Osteogenesis Pathway Analysis Pathway interactions Patients Phenotype Play Polymerase Chain Reaction Procedures Process Production Protein Family Proteins Proto-Oncogenes Publications Publishing RNA RNA Splicing Regulation Replacement Arthroplasty Reverse Transcriptase Polymerase Chain Reaction Role Screening procedure Signal Pathway Signal Transduction Skeletal Development Staging Stem cells Stromal Cells Study models System Systems Biology Techniques Tissue Engineering Tissues Transcript Transcriptase Transcription factor genes Transcriptional Regulation Trypsin USF1 gene USF2 gene Universities Up-Regulation Variant Washington abstracting adapter protein aggrecan aggrecanase angiogenesis articular cartilage base bone cartilage development collagenase design factor C genome-wide in vitro Model in vivo injury and repair link protein lipid biosynthesis malformation mammalian COMP mutant novel overexpression prevent programs promoter repaired research study response response to injury skeletal skeletal disorder stem cell differentiation tool transcription factor vpr Genes

项目摘要

Project Summary/Abstract Osteoarthritis will affect 50 million Americans by 2020. The disease process is characterized by the aberrant gene expression and the inability of cartilage chondrocytes to repair the extracellular matrix leading to cartilage degeneration. CD-RAP is a small cartilage-characteristic protein that is co-expressed in cartilage with type II collagen and aggrecan. It is a compact gene and co-expressed with type IIB collagen in chondrogenesis, making it an excellent model for study of chondrocyte-specific gene expression. Over the past four years, we have extended our knowledge of a single gene to the analysis of the transcriptional regulation of the chondrocyte phenotype. Computational and experimental methods were combined to analyze groups of co- expressed genes and define common regulatory domains of genes expressed in articular cartilage. The current proposal will focus on extending these analyses to define transcriptional regulatory networks that predominantly operate in and define articular cartilage development (as opposed to the growth plate which results in bone formation). As new transcription or signaling factors arise, specific mechanisms of activity will be deciphered using our CD-RAP and COL2A1 gene models or cell culture. The specific aims of the proposal are: (1) Determine the mechanism of transcriptional regulation of the CD-RAP gene by the E-box proteins EF-1/USF1/USF2, C/EBPss, and HMGA. (2) Define the role of transcription factor networks in differentiation of cartilage from stem cells (3) Define the role of transcriptional regulation and signaling in a model for early osteoarthritis and (4) Arising from our preliminary experiments in Specific Aim 3, investigate the role of the intracellular regulator of EGF and FGF signaling, Sprouty-4, in chondrogenesis and osteoarthritis. The experimental approach will focus on (1) using genome-wide expression analyses to determine co-regulated genes, (2) screening for expression of transcription factors using a 1700 oligonucleotide array, (3) computational analyses for transcription factor binding domains of the promoters of co-regulated genes, and (4) confirmation of mechanistic function on the specific cartilage genes, CD-RAP and COL2A1. These studies will apply powerful techniques combining experimental and computational approaches to look beyond candidate genes to elucidate regulatory mechanisms critical for developing strategies to control chondrogenesis during development, in tissue engineering and repair, and ultimately to help find biological methods to control osteoarthritis.

项目概要/摘要到2020年，骨关节炎将影响5000万美国人。疾病过程是其特点是基因表达异常和软骨细胞无能力修复导致软骨退化的细胞外基质。 CD-RAP 是一种小型与 II 型胶原蛋白在软骨中共表达的软骨特征蛋白聚集蛋白聚糖。它是一个紧凑的基因，与 IIB 型胶原蛋白共表达软骨形成，使其成为研究软骨细胞特异性基因的优秀模型表达。在过去的四年里，我们扩展了对单个基因的认识软骨细胞表型的转录调控分析。计算和实验方法相结合来分析共组表达基因并定义表达基因的共同调控域关节软骨。当前的提案将侧重于扩展这些分析来定义转录调控网络主要在特定的细胞中运作并定义软骨发育（与导致骨形成的生长板相反）。随着新的转录或信号传导因子的出现，特定的活性机制将被使用我们的 CD-RAP 和 COL2A1 基因模型或细胞培养物进行破译。具体的该提案的目的是：（1）确定转录调控机制 CD-RAP 基因由 E-box 蛋白 EF-1/USF1/USF2、C/EBPss 和 HMGA 控制。 (2) 定义转录因子网络在软骨与干细胞分化中的作用细胞 (3) 定义转录调控和信号传导在早期模型中的作用骨关节炎和 (4) 由我们在特定目标 3 中的初步实验得出，研究 EGF 和 FGF 信号传导的细胞内调节因子 Sprouty-4 在软骨形成和骨关节炎。实验方法将侧重于（1）使用全基因组表达分析以确定共同调控的基因，（2）筛选使用 1700 个寡核苷酸阵列表达转录因子，(3) 计算分析共调控基因启动子的转录因子结合域， (4) 特定软骨基因 CD-RAP 机械功能的确认和 COL2A1。这些研究将应用强大的技术，结合实验和超越候选基因以阐明调控的计算方法对于制定控制软骨形成策略至关重要的机制开发，组织工程和修复，并最终帮助找到生物控制骨关节炎的方法。