Study of the function of cystatin 10, a novel chondrocyte-specific gene.

研究软骨细胞特异性基因胱抑素 10 的功能。

基本信息

批准号：
14370454
负责人：
TAKESHITA Katsushi
金额：
$ 9.54万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

This study was conducted to elucidate the role and the molecular mechanism of cystatin 10 (Cst10), in the process of endochondral ossification and osteochondral regeneration, and to apply the novel gene, Cst10, to the medical treatment.Cst10 was identified from the mouse auricular cartilage and belongs to the cystatin superfamily, the family of cysteine protease inhibitors. Expression of Cst10 was specific to cartilage, especially to hypertrophic chondrocytes of the growth plate. We found from in vitro studies that Cst10 induced later differentiation and apoptosis of chondrocytes. In order to further investigate the physiological role of Cst10 in vivo, we created mice lacking the Cst10 gene (Cst10KO) and analyzed their bone and cartilage. Cst10KO developed and grew normally without abnormalities of major organs. Radiological and histological analysis revealed an impairment of calcification of the growth plate and a significant decrease in the volume of primary spongiosa adjacent to the … More growth plate by Cst10 deficiency, although bone growth and bone turnover of the Cst10KO remained similar to those of wild type (WT) and heterozygote littermates. In the culture of primary chondrocytes derived from the growth plate, later differentiation of Cst10KO cells was suppressed compared to that of WT cells, which showed the roles of Cst10 expressing in chondrocytes are terminal differentiation and calcification of cartilaginous matrix. In the Cst10KO, calcification was also significantly impaired in other pathological conditions related to endochondral ossification, such as fracture healing and osteophyte formation in osteoarthritis knee model. Furthermore, although WT exhibited calcification of the tendon insertion of patella and calcaneus at 52 weeks of age, these ectopic calcifications were not seen in the Cst10KO. Cst10 was expressed exclusively in type X collagen expressing, matured cells in these calcification regions of WT.These in vitro and in vivo results revealed that Cst10 is an inducer of chondrocyte calcification and contributes to the pathogenesis of osteoarthritis and ectopic calcification without affecting the physiological bone growth or turnover. Less

本研究旨在阐明半胱氨酸蛋白酶抑制剂10（Cst10）在软骨内骨化和骨软骨再生过程中的作用和分子机制，并将新基因Cst10在医学治疗中的应用。Cst10从小鼠耳廓中分离出来。软骨，属于半胱氨酸蛋白酶抑制剂家族，Cst10 的表达对软骨具有特异性。特别是对生长板的肥大软骨细胞，我们通过体外研究发现Cst10诱导软骨细胞的后期分化和凋亡。为了进一步研究Cst10在体内的生理作用，我们创建了缺乏Cst10基因的小鼠（Cst10KO）并进行了分析。他们的骨骼和软骨发育和生长正常，主要器官没有异常，放射学和组织学分析显示生长板钙化受损并显着减少。尽管 Cst10KO 的骨生长和骨转换与野生型 (WT) 和杂合子同窝幼体的原代软骨细胞培养物相似，但 Cst10 缺陷导致邻近生长板的原代海绵体体积增加。平板中，与WT细胞相比，Cst10KO细胞的后期分化受到抑制，这表明软骨细胞中表达的Cst10的作用是软骨的终末分化和钙化在 Cst10KO 中，与软骨内骨化相关的其他病理状况（例如骨关节炎膝关节模型中的骨折愈合和骨赘形成）中的钙化也显着受损。此外，尽管 WT 在 52 周时表现出髌骨和跟骨的肌腱附着点钙化。随着年龄的增长，这些异位钙化在 Cst10KO 中未见，Cst10 仅在表达 X 型胶原的成熟细胞中表达。 WT 的这些钙化区域。这些体外和体内结果表明，Cst10 是软骨细胞钙化的诱导剂，有助于骨关节炎和异位钙化的发病机制，而不影响生理性骨生长或更新。