Study on signal transduction in osteoclastogenesis for the development of anti-osteoporosis drugs.

破骨细胞生成信号转导研究，用于抗骨质疏松药物的开发。

• 批准号: 11557139
• 负责人: TAKAHASHI Naoyuki
• 金额: $ 8.64万
• 依托单位: Showa 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 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557139/
• 关键词: Osteoclast; Osteoblast; Inflammatory cytokines; Calcium; BMP; RANKL; OPG; Osteoporosis; ODF; RANK; TNFα; IL-1; 細胞内Ca

Inhibition of signal transduction for osteoclastogenesis is thought to provide important information for development of anti-osteoporosis drugs. We have succeeded in molecular cloning of an essential factor for osteoclastogenesis, RANKL, which is expressed as a membrane associated factor in osteoblasts in response to many bone-resorbing factors. Using culture systems for osteoclast formation, we have studied signal transduction for osteoclastogenesis including RANK-RANKL signaling. The findings obtained from the study are as follows. (1) TNFα stimulated osteoclastogenesis in bone marrow-derived macrophage cultures in the presence of M-CSF.(2) SaOS-4/3, a subclone of the human osteosarcoma cell line SaOS-2, established by transfecting the human PTH/PTHrP receptor cDNA, supported osteoclast formation in response to PTH in co-culture with mouse bone marrow cells. Expression of mRNAs for RANKL and M-CSF by SaOS-4/3 cells was up-regulated in response to PTH.Both factors were expressed as membrane-associated factors. (3) Ionomycin and A23187 stimulated osteoclast formation in mouse co-cultures of bone marrow cells and osteoblasts. We also found that PMA, an activator of protein kinase C, stimulated osteoclast formation in the co-culture though up-regulation of RANKL expression in osteoblasts. (4) BMP-2 markedly enhanced osteoclast differentiation induced by RANKL and M-CSF.Addition of a soluble form of BMP receptor type-IA to the culture inhibited not only osteoclast formation induced by RANKL and BMP-2, but also the basal osteoclast formation supported by RANKL alone. Both bone marrow macrophages and mature osteoclasts expressed BMP-2 and BMP receptor type IA mRNAs. These findings provide important information for the development of anti-osteoporosis drugs.

破骨细胞生成信号转导的抑制被认为为抗骨质疏松药物的开发提供了重要信息，我们已经成功地分子克隆了破骨细胞生成的必需因子RANKL，该因子在成骨细胞中表达为响应许多骨的膜相关因子。 -再吸收因子。使用破骨细胞形成的培养系统，我们研究了破骨细胞生成的信号转导，包括RANK-RANKL信号转导，从研究中获得的结果如下。 (1) 在 M-CSF 存在的情况下，TNFα 刺激骨髓源性巨噬细胞培养物中的破骨细胞生成 (2) SaOS-4/3，人骨肉瘤细胞系 SaOS-2 的亚克隆，通过转染人 PTH/PTHrP 建立。受体 cDNA，支持破骨细胞形成，以响应与小鼠骨髓细胞共培养的 PTH，SaOS-4/3 表达 RANKL 和 M-CSF 的 mRNA。细胞对 PTH 的反应上调。这两种因子均表达为膜相关因子 (3) 离子霉素和 A23187 在小鼠骨髓细胞和成骨细胞共培养物中刺激破骨细胞形成。蛋白激酶 C 通过上调成骨细胞中的 RANKL 表达刺激共培养物中的破骨细胞形成 (4) BMP-2 显着增强了由成骨细胞诱导的破骨细胞分化。 RANKL和M-CSF。在培养物中添加可溶形式的BMP受体IA型不仅抑制RANKL和BMP-2诱导的破骨细胞形成，而且抑制仅由RANKL支持的基础破骨细胞形成和成熟的骨髓巨噬细胞。破骨细胞表达 BMP-2 和 IA 型 BMP 受体 mRNA，这些发现为抗骨质疏松药物的开发提供了重要信息。

项目成果

Jimi, E., Akiyama, S., Tsurukai, T., Okahashi, N., Kobayashi, K., Udagawa, N., Nishihara, T., Takahashi, N., Suda T.: "Osteoclast differentiation factor (ODF) acts as a multifunctional regulator in murine osteoclast differentiation and function."J.Immunol

Jimi，E.，Akiyama，S.，Tsurukai，T.，冈桥，N.，小林，K.，宇田川，N.，西原，T.，高桥，N.，须田T.：“破骨细胞分化因子（ODF）

Itoh K, Udagawa N, Katagiri T, Iemura S, Ueno N, Yasuda H, Higashio K, Quinn JMW, Gillespie MT, Martin TJ, Suda T & Takahashi N.: "Bone morphogenetic protein 2 stimulates osteoclast differentiation and survival supported by receptor activator of nuclear f

伊藤 K、宇田川 N、片桐 T、伊村 S、上野 N、安田 H、东尾 K、奎因 JMW、吉莱斯皮 MT、马丁 TJ、须田 T

Takami, M., Takahashi, N., Udagawa, N., Miyaura, C., Suda, K., Woo, J.-T., Martin, T.J., Nagai, K., Suda, T.: "Intracellular calcium and protein kinase C mediate expression of receptor activator of NF-κB ligand (RANKL) and osteoprotegerin in osteoclasts."

Takami, M.、Takahashi, N.、Udakawa, N.、Miyaura, C.、Suda, K.、Woo, J.-T.、Martin, T.J.、Nagai, K.、Suda, T.：“细胞内钙蛋白激酶 C 介导破骨细胞中 NF-κB 配体受体激活剂 (RANKL) 和骨保护素的表达。”

Itoh K, et al.: "BMP-2 stimulates osteoclast differentiation and survival supported by RANKL."Endocrinology. (in press). (2001)

Itoh K 等人：“BMP-2 刺激破骨细胞分化和存活，并由 RANKL 支持。”内分泌学。

Itoh, K., Udagawa, N., Matsuzaki, K., Takami, M., Amano, H., Shinki, T., Ueno, Y., Takahashi, N., Suda, T.: "Importance of membrane-or matrix-associated forms of M-CSF and RANKL/ODF in osteoclastogenesis supported by SaOS-4/3 cells expressing recombinant

Itoh, K.、Udakawa, N.、Matsuzaki, K.、Takami, M.、Amano, H.、Shinki, T.、Ueno, Y.、Takahashi, N.、Suda, T.：“膜的重要性-