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Regulation of cartilage destruction using tissue-specific expression system

使用组织特异性表达系统调节软骨破坏

基本信息

批准号：
11470305
负责人：
KIMURA Tomoatsu
金额：
$ 7.36万
依托单位：
TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470305/
关键词：
cartilage repair gene collagen

项目摘要

The present study can be summarized into four categories ; analysis of genes responsible for cartilage damage, analysis of tissue-specific element of Col11a2 gene, prevention of cartilage damage and its repair. Firstly, we demonstrated that aggrecan gene VNTR in the core protein coding region and VDR allele are associated with the degeneration of intervertebral disc cartilage. We then identified cartilage tissue specific cis-element of Col11a2 and used this element for the tissue specific expression of marker gene after in vivo gene transfer. To further improve such gene transfection efficiency we developed a new superfine-pointed 7-needle electrodes for in vivo electroporetion. Bearing these background results, we then prepared engineered mesenchymal cells from the bone marrow (MSCs) by transfecting cartilage-derived morphogenetic protein 1 (CDMP1). The cells were used for transplantation into full-thickness articular cartilage defect in rabbit knee joint and the specimens were evaluated histologically. During in vivo repair of articular cartilage defect, cartilage regeneration was significantly enhanced with transplantation of CDMP1-transfected MSCs. The defect was filled with hyaline cartilage and the deeper zone was remodeled to subchondral bone with time. The histological score of CDMP1-transfected MSCs group was better than those of control MSCs group and empty control. The results demonstrated that CDMP1-transfected MSCs maintained cell growth activity, showed enhanced progression toward chondrogenic lineage and improved the in vivo cartilage repair.

目前的研究可归纳为四个方面：软骨损伤相关基因的分析、Col11a2基因组织特异性元件的分析、软骨损伤的预防与修复。首先，我们证实了核心蛋白编码区的聚集蛋白聚糖基因VNTR和VDR等位基因与椎间盘软骨退变相关。然后我们鉴定了Col11a2的软骨组织特异性顺式元件，并将该元件用于体内基因转移后标记基因的组织特异性表达。为了进一步提高基因转染效率，我们研制了一种新的用于体内电穿孔的超细尖七针电极。基于这些背景结果，我们通过转染软骨衍生形态发生蛋白1（CDMP 1）从骨髓中制备了工程化间充质细胞（MSCs）。将其移植于兔膝关节全层软骨缺损处，并进行组织学观察。在关节软骨缺损的体内修复过程中，移植CDMP1转染的MSCs可显著促进软骨再生。缺损区填充透明软骨，随着时间的推移，更深的区域重塑为软骨下骨。CDMP 1转染MSCs组的组织学评分优于对照MSCs组和空白对照组。结果表明，转染CDMP 1的MSCs保持细胞生长活性，显示出向软骨细胞谱系的增强进展，并改善体内软骨修复。