Analysis of chondrodysplasias using transgenic mouse model.

使用转基因小鼠模型分析软骨发育不良。

• 批准号: 03454362
• 负责人: KIMURA Tomoatsu
• 金额: $ 4.03万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03454362/
• 关键词: cartilage; collagen; transgenic mouse; osteoarthritis; 遺伝子; コラ-ゲン

Type IX collagen is a hybrid extracellular matrix component, with the properties of both a proteoglycan and a collagen. It is found in hyaline cartilage, vitreous humor and embryonic cornea, where it is located along the surfaces of type II-containing collagen fibrils. To investigate the functional role of type IX collagen in cartilage, we have introduced an alphal(IX) gene construct expressing a truncated alphal(IX) chain into mice. The construct was designed with an in frame deletion in the central part of alphal(IX) cDNA and tissue specific expression in cartilages and eyes was obtained by using the type II collagen promoter-enhancer. In mouse tissues the transgene was expressed as shortened alphal(IX) chains that were disulfide-bonded with endogenous polypeptides to form collagenous molecules. Light and electron microscopic examination of the offspring of three different founders revealed pathological changes similar to osteoarthritis in the articular cartilage of knee joints. In addition, mice homozygous for the transgene developed mild chondrodysplasia ; i.e., mild dwarfism, anterior tonguing in the vertebral bodies, and ophthalmopathy. The relative ratio of transgene product to the endogenous alphal(IX) chain was approximately one in homozygotes and less than one in heterozygotes. Therefore, the phenotypic severity correlated well with the level of transgene expression. Given these findings, it is possible that mutations in type IX collagen genes may cause certain forms of chondrodysplasia and osteoarthritis in humans.

IX型胶原蛋白是一种混合细胞外基质组分，具有蛋白聚糖和胶原蛋白的性质。在透明软骨、玻璃体液和胚胎角膜中发现，其位于沿着含II型胶原纤维的表面。为了研究IX型胶原在软骨中的功能作用，我们将表达截短的IX链的IX基因构建体引入小鼠。该构建体被设计为在软骨（IX）cDNA的中心部分具有框内缺失，并且通过使用II型胶原启动子-增强子获得软骨和眼睛中的组织特异性表达。在小鼠组织中，转基因表达为缩短的半乳糖醛（IX）链，其与内源性多肽二硫键结合形成胶原分子。光镜和电镜检查的后代的三个不同的创始人揭示了病理变化类似于骨关节炎的关节软骨的膝关节。此外，转基因纯合子小鼠出现轻度软骨发育不良;即，轻度侏儒症、椎体前舌和眼病。转基因产物与内源性DNA（IX）链的相对比值在纯合子中约为1，在杂合子中小于1。因此，表型严重程度与转基因表达水平相关性良好。鉴于这些发现，IX型胶原基因的突变可能会导致人类某些形式的软骨发育不良和骨关节炎。

项目成果

M.Iwasaki,et al.: "Transforming growth factor-β1 stimulates chain drogenesis and inhibits osteogenesis in high density culture of periosteum-derived cells." Endocrinology. 132. 1603-1608 (1993)

M.Iwasaki 等人：“转化生长因子-β1 刺激骨膜来源细胞的高密度培养物中的链生成并抑制成骨。” 132. 1603-1608 (1993)。

M.Iwasaki, et al.: "Transforming growth factor-beta1 stimulates chondrogenesis and inhibits osteogenesis in high density culture of periouteum-derived cells." Endocrinology. 132. 1603-1608 (1993)

M.Iwasaki 等人：“转化生长因子-β1 在高密度培养的膜衍生细胞中刺激软骨形成并抑制成骨。”

T.Kimura,et al.: "Altered chondrocyte gene expression in articular cartilage matrix components and susceptibility to cartilage destruction" In:Cell Mechanics and Cellular Engineering,Ed.,V.C.Mow.Springer-Verlag,New York(in press),

T.Kimura 等人：“关节软骨基质成分中软骨细胞基因表达的改变和对软骨破坏的敏感性”，见：细胞力学和细胞工程，编辑，V.C.Mow.Springer-Verlag，纽约（正在印刷中），

K.Nakata,at al.: "Ostcocrothintis associated with unild choondrodysplasiab in transgenic mice expressing α1 (IX) collagen chaius with a central delction" Proc.Natl.Acad.Sci.USA. 90. 2870-2874 (1993)

K.Nakata 等人：“表达 α1 (IX) 胶原链的转基因小鼠中与软骨发育不良相关的骨软骨炎”Proc.Natl.Acad.Sci.USA 90. 2870-2874 (1993)。

T.Kimura, et al.: Altered chondrocyte gene expression in articular cartilage matrix components and susceptibility to cartilage destruction. in Cell Mechanics and Cellular Engineering (Eds.V.C.Mow, et al.). Springer-Verlag, New York, 445-456

T.Kimura 等人：改变关节软骨基质成分中的软骨细胞基因表达和对软骨破坏的易感性。