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，在干细胞分化过程中起着至关重要的作用。它在间质组织形成中的作用也有牵连，尽管详细的机制尚未分析。为了阐明Notch信号在成骨过程中的功能，我们将构成活性的Notch （Notch胞内结构域，NICD）转染到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）来检测Notch信号。NICD普遍抑制了Kusa-Al和KusaO中成骨标志物基因的表达、钙沉积、体外矿化、Cbfal和Ose2元件的启动子活性。NICD显著抑制了Kusa-Al的体内骨形成。这些结果提示Notch信号在成骨细胞分化中起负向调节作用。为了证实这些结果，Notch信号被一种进化中更保守的转录因子CBF1（也称为RBP-Jic）抑制。短暂转染CBF1显著提高了Ose2和Cbfal启动子活性，但阻断了NICD转染诱导的Hesi启动子活性的升高。在稳定表达CBF1的Kusa-AI/CBF1细胞系中，钙沉积、体外矿化和RANKL表达等成骨特性显著提高。kasa - a1 /CBFI显著促进了体内硬组织的形成。此外，与kasa - a1 /CBF1相比，kasa - a1 /CBF1形成的硬组织结构更紧凑，类似于骨小梁组织。CBF1的表达普遍存在，但在骨组织和骨形成结构中表达尤其高。这些结果表明，转录介质CBF1的过表达强烈促进了间充质祖细胞的成骨分化。综上所述，Notch信号通路在成骨细胞分化过程中起着至关重要的作用，而Notch信号通路的调控是调控成骨细胞矿化的关键。少

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。