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.

项目摘要/摘要到2020年，骨关节炎将影响5000万美国人。疾病过程是以异常基因表达和软骨软骨细胞的无能为特征修复细胞外基质，导致软骨变性。 CD-Rap很小软骨特征蛋白与II型胶原蛋白和软骨共表达 Aggrecan。它是一个紧凑的基因，并与IIB型胶原蛋白共表达软骨发生，使其成为研究软骨细胞特异性基因的绝佳模型表达。在过去的四年中，我们扩展了对单个基因的了解分析软骨细胞表型的转录调节。合并了计算和实验方法，以分析共同表达的基因并定义了在关节软骨。当前的提案将着重于扩展这些分析以定义转录调节网络主要运行并定义关节软骨发育（与导致骨形成的生长板相反）。随着新的转录或信号传导因素的出现，活动的具体机制将是使用我们的CD-RAP和COL2A1基因模型或细胞培养的解密。具体该提案的目的是：（1）确定转录调节的机制 E-Box蛋白EF-1/USF1/USF2，C/EBPS和HMGA的CD-RAP基因。（2）定义转录因子网络在软骨与茎分化中的作用细胞（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）是由我们在特定目标3中的初步实验引起的研究EGF和FGF信号传导的细胞内调节因子在Sproty-4中的作用软骨发生和骨关节炎。实验方法将重点放在（1）使用全基因组表达分析以确定共同调节的基因，（2）筛选使用1700寡核苷酸阵列的转录因子表达，（3）计算共同调节基因启动子的转录因子结合结构域的分析，（4）确认特定软骨基因的机械功能，CD-RAP 和col2a1。这些研究将采用结合实验和超越候选基因以阐明调节的计算方法对于制定控制软骨发生策略至关重要的机制开发，组织工程和维修，并最终有助于找到生物学控制骨关节炎的方法。