TRANSCRIPTIONAL REGULATORS OF CHONDROINDUCTION BY DBP

DBP 软骨诱导的转录调节因子

• 批准号: 6677579
• 负责人: KAREN E YATES
• 金额: $ 17.3万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6677579
• 关键词: cartilage development; cell differentiation; cell growth regulation; child physical development; chondrocytes; fibroblasts; gene expression; human tissue; newborn human (0-6 weeks); oligonucleotides; protein localization; tissue /cell culture; transcription factor

DESCRIPTION (provided by applicant): There is a need for innovative, effective cartilage repair techniques. Limited knowledge of the molecular mechanisms that regulate postnatal chondroblast differentiation (chondroinduction) confounds the design of better repair strategies. This exploratory/developmental project from a New Investigator tests the hypothesis that transcriptional regulator(s) of postnatal chondrogenesis differ from those of prenatal chondrogenesis. The project uses a novel, in vitro system for postnatal chondroinduction that models the conversion of mesenchymal cells into chondroblasts by demineralized bone powder (DBP) in vivo. Human dermal fibroblasts (hDFs) cultured in porous, 3-dimensional collagen sponges with DBP develop a chondroblast phenotype after 7 days, as shown by quantitative biochemical, immunochemical, and gene expression assays for cartilage matrix components. This experimental system is highly relevant to human health, as DBP is used clinically in periodontal, craniomaxillofacial, and orthopedic surgery applications. Previously, we showed that DBP induces specific shifts in gene expression by day 3, prior to full expression of the chondroblast phenotype. The altered expression of several functional classes of genes suggests that DBP activates transcriptional regulators in target cells. Therefore, Specific Aim I is to characterize the expression and activity of transcription factors during postnatal chondroinduction by DBP. Specific genes of interest include transcription factors found to be changed by DBP on day 3 in our Preliminary Studies (ID2, FOG2, MBLL, HBP1), and transcription factors that are known to regulate embryonic chondrogenesis (SOX9, HOXC8, scleraxis). The transcription factor expression profile of postnatal chondrogenesis will be compared to the known patterns of expression during prenatal chondrogenesis. In Specific Aim II, we will determine whether RDA31, a putative zinc-finger protein whose expression is increased in chondroinduced hDFs on day 3, has properties expected of a transcription factor (nuclear localization and transcriptional activator activity). Specific Aim III is to elucidate the transcriptional mechanisms that regulate postnatal chondrogenesis by DBP. We will determine which DBP-regulated transcription factors are necessary and/or sufficient for postnatal chondrogenesis. This proposed work would have a high impact of design of strategies for repair and/or regeneration of cartilage.

描述（由申请人提供）：需要创新，有效的软骨维修技术。对调节产后软骨细胞分化（软骨诱导）的分子机制的了解有限，这会使更好的维修策略的设计混淆。这项来自新研究者的探索性/发育项目检验了以下假设：产后软骨发生后的转录调节剂与产前软骨发生不同。该项目使用一种新型的体外系统来用于产后软骨诱导，该系统通过体内脱源性骨粉（DBP）将间充质细胞转化为软骨细胞的转化。 7天后，用DBP培养的人类真皮成纤维细胞（HDFS）在多孔的三维胶原海绵中培养的是软骨细胞表型，如定量生化，免疫化学，免疫化学，免疫化学，和基因的基因表达分析，用于软骨矩阵组件。该实验系统与人类健康高度相关，因为DBP在牙周，颅面和骨科手术应用中用于临床上。以前，我们表明DBP在第3天之前诱导基因表达的特异性移位，然后在软骨细胞表型的完全表达之前。几种功能类别基因的表达改变表明DBP激活靶细胞中的转录调节剂。因此，特定的目的是表征DBP产后软骨诱导期间转录因子的表达和活性。感兴趣的特定基因包括在我们的初步研究（ID2，FOG2，MBLL，HBP1）中发现DBP改变的转录因子，以及已知会调节胚胎软骨发生的已知转录因子（SOX9，HOXCC8，Scleraxis）。在产后软骨发生期间，将将出生后软骨发生的转录因子表达谱进行比较。在特定的目标II中，我们将确定RDA31是一种推定的锌指蛋白，其表达在第3天的软骨诱导的HDF中的表达是否增加，它具有转录因子（核定位和转录激活剂活性）的性能。具体目的III是阐明DBP调节产后软骨发生的转录机制。我们将确定哪些DBP调节的转录因子是必需的和/或足以用于产后软骨发生。这项拟议的工作将对软骨修复和/或再生的策略设计产生很大的影响。