Osteoblast Differentiation and Notch Signaling

成骨细胞分化和Notch信号传导

• 批准号: 14370615
• 负责人: KAWASHIMA Nobuyuki
• 金额: $ 7.42万
• 依托单位: Tokyo Medical and Dental University
• 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 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370615/
• 关键词: Notch; GBF1; Osteoblasts; Osteogenesis; Mesenchymal stem cell; Kusa; Mineralization; Hes; 骨髄間葉細胞; KusaA1; 硬組織形成; 骨髄ストローマ細胞; 分化; オステオカルシン; ALPase; 幹細胞

Notch is a trausmembrane receptor that plays a crucial role in differentiation of stem cells. Its role in the formation of mesenchymal tissues has also been implicated although detailed mechanism has not yet been analyzed. To elucidate the function of Notch signaling in osteogenesis, the constitutively active Notchi (Notch intracellular domain, NICD) was transfected into two different osteoblastic mesenchymal cell lines, Kusa-Al and Kusa-O, and established stable transformants (KusaANICD and KusaONICD) to examine the Notch signaling. NICD generally suppressed the expression of osteogenic marker genes, calcium deposition, in vitro mineralization, the promoter activities of the Cbfal and the Ose2 element in both Kusa-Al and KusaO. In vivo bone formation of Kusa-Al was significantly suppressed by NICD. These results imply that Notch signaling functions as a negative regulator on osteoblast differentiation. In order to confirm these results, the Notch signaling was suppressed by an evoluti … More onarily conserved transcription factor CBF1, also known as RBP-Jic. Transient transfection of CBF1 remarkably increased the promoter activities of Ose2 and Cbfal, but blocked the elevation of Hesi promoter activity induced by NICD transfection. In Kusa-AI/CBF1 cell line, which is a CBF1 expressing stable cell line, the osteogenic properties, including calcium deposition, in vitro mineralization and the RANKL expression were significantly promoted. The in vivo hard tissue formation was greatly facilitated in Kusa-A1/CBFI. Furthermore, the hard tissues formed by Kusa-A1/CBF1 showed more compact structure similar to trabecular bone tissues than those by Kusa-AI/host. The expression of CBF1 was ubiquitous, but it was especially high in bone tissues and bone forming structures. These, results indicated that over-expression of transcription mediator CBF1 strongly promoted the osteogenic differentiation of mesenchymal progenitor cells. Taken together, the Notch signaling should be essential in the osteoblastic differentiation, and modification of its signaling would be key to control the mineralization. Less

Notch 是一种跨膜受体，在干细胞的分化中发挥着至关重要的作用，但其在间充质组织形成中的作用也受到影响，但尚未分析其详细机制。将Notchi（Notch胞内结构域，NICD）转染至两种不同的成骨间充质细胞系Kusa-Al和Kusa-O中，并建立稳定的转化子（KusaANICD 和 KusaONICD）检查了 Notch 信号传导在 Kusa-Al 和 KusaO 体内骨形成中普遍抑制成骨标记基因、钙沉积、体外矿化、Cbfal 和 Ose2 元件的启动子活性。 Kusa-Al 的表达被 NICD 显着抑制。这些结果表明 Notch 信号传导对成骨细胞分化起到负调节作用。为了证实这些结果，Notch 信号传导被抑制。在 Kusa-AI/ 中，通过进化保守的转录因子 CBF1（也称为 RBP-Jic），CBF1 的瞬时转染显着增加了 Ose2 和 Cbfal 的启动子活性，但阻止了 NICD 转染诱导的 Hesi 启动子活性的升高。 CBF1细胞系是表达CBF1的稳定细胞系，其成骨特性，包括钙沉积、体外矿化和体内RANKL表达均得到显着促进。 Kusa-A1/CBFI 大大促进了硬组织的形成，此外，Kusa-A1/CBF1 形成的硬组织比 Kusa-AI/宿主形成的硬组织表现出更致密的结构，类似于骨小梁组织。CBF1 的表达无处不在。但它在骨组织和骨形成结构中尤其高，这些结果表明转录介质 CBF1 的强烈过度表达促进了间充质祖细胞的成骨分化。 Notch 信号传导在成骨细胞分化中至关重要，对其信号传导的修饰将是控制矿化的关键。

项目成果

Shindo K., Kawashima N., Sakamoto K., Umezawa A., et al.: "Osteogenic differentiation of the mesenchymal progenitor cells, Kusa is suppressed by Nothc signaling."Experimental Cell Research. 290. 370-380 (2003)

Shindo K.、Kawashima N.、Sakamoto K.、Umezawa A. 等人：“间充质祖细胞 Kusa 的成骨分化受到 Nothc 信号传导的抑制。”实验细胞研究。

Shindo K., Kawashima N., Sakamoto K., Umezawa A., et al.: "Osteogenic differentiation of the mesenchymal progenitor cells, Kusa is suppressed by Notch signaling."Experimental Cell Research. 290. 370-380 (2002)

Shindo K.、Kawashima N.、Sakamoto K.、Umezawa A. 等人：“间充质祖细胞 Kusa 的成骨分化受到 Notch 信号传导的抑制。”实验细胞研究。

Shindo K., Kawashima N., Sakamoto K., Umezawa A., et al.: "Osteogenic differentiation of the mesenchymal progenitor cells, Kusa is suppressed by Notch signaling."Experimental Cell Research. 290. 370-380 (2003)

Shindo K.、Kawashima N.、Sakamoto K.、Umezawa A. 等人：“间充质祖细胞 Kusa 的成骨分化受到 Notch 信号传导的抑制。”实验细胞研究。