Functional analysis of TNF signaling in bone metabolism using knocked-out mice of type I and II TNF receptors.

使用 I 型和 II 型 TNF 受体敲除小鼠对骨代谢中 TNF 信号传导进行功能分析。

• 批准号: 14571385
• 负责人: ENOMOTO Hiroshi
• 金额: $ 2.24万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571385/
• 关键词: TNF receptor; knock-out mice; osteoclasts; 破骨細胞; 骨形態計測; pQCT

TNF signaling is known to play a crucial role in bone absorption under inflammatory condition. To distinguish the functional role of type I and type II TNF receptor, knock-out mice of each receptor were subjected to histological and bone morphometric analysis. In 12 wks old femurs, bone volume, thickness and mineral content in both cortical and cancellous bone were decreased in type I (-/-) mice, whereas these parameters were increased in type II (-/) mice. To investigate a molecular mechanism of TNF receptor involved bone metabolism, osteoblasts and osteoclasts were cultured from calvariae and macrophage from bone marrow, respectively. Treatment of osteoblasts with BMP-2 or Dexamethasone, however, did not alter their ALP expression as well as cell proliferation. In contrast, osteoclast formation was only induced in type II (-/-) macrophage by treating with M-CSF and RANKL, indicating that certain molecules secreted of exposed on cell surface could be important for osteoclasts formation. To identify these molecules, we applied DNA tip analysis to each cultured cell and found three novel and 5 known genes that could be target or down-stream signaling for TNF receptor activation. We are now analyzing the functional role and expression pattern of these genes.

已知TNF信号传导在炎症条件下的骨吸收中起关键作用。为了区分I型和II型TNF受体的功能作用，对每种受体的敲除小鼠进行组织学和骨形态测定分析。在12周龄股骨中，I型（-/-）小鼠皮质骨和松质骨的骨体积、厚度和矿物质含量均降低，而II型（-/）小鼠这些参数均升高。为探讨肿瘤坏死因子受体参与骨代谢的分子机制，分别从颅骨培养成骨细胞和骨髓培养巨噬细胞。然而，用BMP-2或地塞米松处理成骨细胞并没有改变它们的ALP表达以及细胞增殖。与此相反，破骨细胞的形成仅诱导II型（-/-）巨噬细胞与M-CSF和RANKL处理，表明某些分子分泌或暴露在细胞表面上可能是破骨细胞的形成重要。为了鉴定这些分子，我们对每个培养的细胞进行了DNA尖端分析，发现了3个新的和5个已知的基因，这些基因可能是TNF受体激活的靶基因或下游信号转导基因。我们现在正在分析这些基因的功能作用和表达模式。

项目成果

Iioka et al.: "p300/CBP Acts as a Coactivator to Cartilage Homeoorotein-1(Cart1),Paired-Like Homeoprotein, via Acetylation of the Conserved Lysine Residue Adjacent to the Homeodomain"J Bone Miner Res. 18・8. 1419-1429 (2003)

Iioka 等人：“p300/CBP 通过同源结构域附近保守赖氨酸残基的乙酰化，充当软骨同源蛋白-1 (Cart1) 的共激活剂”J Bone Miner Res 18・8。 1429 (2003)

Iioka et al.: "p300/CBP Acts as a Coactivator to Cartilage Homeoprotein-1 (Cart1), Paired-Like Homeoprotein, via Acetylation of the Conserved Lysine Residue Adjacent to the Homeodomain"J Bone Miner Res. (In press). (2003)

Iioka 等人：“p300/CBP 通过与同源结构域相邻的保守赖氨酸残基乙酰化，充当软骨同源蛋白-1 (Cart1)（配对样同源蛋白）的共激活剂”J Bone Miner Res。

Hirata et al.: "Transnlantation of skin fibroblasts expressing BMP-2 promotes Bone repair more effectively than those expressing Runx2"BONE. 32・5. 502-512 (2003)

Hirata 等人：“表达 BMP-2 的皮肤成纤维细胞的移植比表达 Runx2 的皮肤成纤维细胞更有效地促进骨修复”BONE 32·5 (2003)。

Furukawa et al.: "Functional domains of paired-like homeoprotein Cart1 and the relationship between dimerization and transcription activity"Genes to Cells. 7. 1135-1147 (2002)

Furukawa 等人：“配对样同源蛋白 Cart1 的功能域以及二聚化和转录活性之间的关系”基因到细胞。

Hirata et al.: "Transplantation of skin fibroblasts expressing BMP-2 promotes Bone repair more effectively than those expressing Runx2"BONE. (in press). (2003)

Hirata 等人：“移植表达 BMP-2 的皮肤成纤维细胞比表达 Runx2 的皮肤成纤维细胞更有效地促进骨修复”BONE。