Analysing the Role of Growth Plate Specific Genes in Endochondral Ossification by Specific Overexpression or Deletion under the BAC Col10a1 Promoter

通过 BAC Col10a1 启动子下的特异性过表达或缺失分析生长板特异性基因在软骨内骨化中的作用

• 批准号: 36677775
• 负责人: Professor Dr. Klaus von der Mark, Ph.D.
• 金额: --
• 依托单位: Lehrstuhl für Experimentelle Medizin I - Molekulare Pathogeneseforschung
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/36677775?language=en
• 关键词: Analysing; Role; Growth; Plate; Specific

Cartilage-bone transition by endochondral ossification is a highly complex process which determines not only longitudinal growth of long bones, ribs and vertebrae, but also plays a critical role in bone fracture callus healing, osteophyte formation and cartilage tissue engineering. Decisive steps in this process are maturation and hypertrophy of chondrocytes in the growth plate before inducing mineralisation, apoptosis and resorption of hypertrophic cartilage followed by bone marrow invasion. These events are regulated by synergistic interactions of several signalling pathways and transcriptions factors controlled by a multitude of growth factors and hormones. In order to elucidate specific functions of genes presumably involved in endochondral ossification, we designed a collagen X specific BAC (Bacterial Artificial Chromosome) recombineering system which allows specific and robust overexpression of reporter genes and other genes of interest in hypertrophic cartilage under the authentic Co110a1 promoter in transgenic mice. The specific aim of this project is to generate a Co110a1-BAC-CRE mouse which will allow specific gene deletion in the growth plate of mutated mice by removing exons flanked by 1oxP sites, primarily (l integral, PTHrP and (-catenin. Second, the transcription factor Sox9 and the hormone-like factor PTHrP, both know to delay chondrocyte maturation, will be overexpressed in BAC –Co110a1 transgenic mice in order to elucidate their specific function in endochondral ossification. We propose that this Co110a1-specific BAC recombineering system will provide a powerful and specific new tool to elucidate and to manipulate the complex process of endochondral ossification.

软骨内成骨转变是一个高度复杂的过程，不仅决定了长骨、肋骨和椎骨的纵向生长，而且在骨折痂愈合、骨赘形成和软骨组织工程中起着至关重要的作用。这一过程的决定性步骤是生长板中软骨细胞的成熟和肥大，然后诱导矿化、细胞凋亡和肥厚软骨的再吸收，然后是骨髓侵袭。这些事件是由多种生长因子和激素控制的几种信号通路和转录因子的协同相互作用调节的。为了阐明可能参与软骨内成骨的基因的特定功能，我们设计了一个胶原X特异性BAC（细菌人工染色体）重组系统，该系统允许在转基因小鼠的真实Co110a1启动子下特异性和稳健地过表达肥大软骨的报告基因和其他感兴趣的基因。该项目的具体目标是生成Co110a1-BAC-CRE小鼠，通过去除1oxP位点两侧的外显子，主要是(l - integral， PTHrP和（-catenin），可以在突变小鼠的生长板中删除特定的基因。其次，已知具有延缓软骨细胞成熟的转录因子Sox9和激素样因子PTHrP将在BAC -Co110a1转基因小鼠中过表达，以阐明其在软骨内成骨中的特定功能。我们认为，这种co110a1特异性BAC重组系统将为阐明和操纵软骨内成骨的复杂过程提供一个强大而特异性的新工具。