Roles of Notch and Nov signaling on regeneration of hard tissues and a development of their clinical application

Notch和Nov信号在硬组织再生中的作用及其临床应用进展

• 批准号: 16390543
• 负责人: KAWASHIMA Nobuyuki
• 金额: $ 8.77万
• 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: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390543/
• 关键词: osteoblasts; Notch; Nov; mesenchymal cells; differentiation; mineralization; CBF1; RBP-Jκ; Kusa-A1; 硬組織形成; 骨髄ストローマ細胞; KusaA1; 骨髄ストローマ

Notch is a transmembrane 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, we transfected the constitutively active Notch 1 (Notch intracellular domain, NICD) into two different osteoblastic mesenchymal cell lines, Kusa-A1 and KusaO, and established stable transformants (KusaA1NICD and KusaONICD) to examine the Notch signaling. NICD generally suppressed the expression of osteogenic marker genes, calcium deposition and in vitro mineralization in both Kusa-A1 and KusaO. The promoter activities of the Cbfa1 and the Ose2 element were attenuated by NICD. In vivo bone formation of Kusa-A1 was significantly suppressed by NICD. These results suggest that Notch signaling functions as a general coordinator of osteogenesis through cell-cell interactions.Nephroblastoma overexpressed gene (Nov : CCN3) is a cysteine-rich protein that is overexpressed in avian nephroblastomas, and it is thought to be involved in the control of cell proliferation and development of various tissues. As Nov has recently reported to bind Notch, we evaluated the Nov signaling in the osteoblastic mesenchymal cell lines. Transfection of Nov expression vector to Kusa-A1 induced downregulation of osteoblastic markers. On the centrally, Notch related genes were upregulated. DNA microarray revealed that Nov expression was highly induced in KusaA1NICD. Therefore, Nov may induce the Notch signaling, which further induced Nov expression, and this cycle may be essential to keep the cells in immature condition.

Notch是一种跨膜受体，在干细胞的分化中起着至关重要的作用。尽管尚未分析详细机制，但它在间充质组织形成中的作用也被暗示。为了阐明Notch信号在成骨中的功能，我们将组成型活性Notch 1（Notch细胞内结构域，NICD）转染到两种不同的成骨间充质细胞系Kusa-A1和KusaO中，并建立稳定的转化体（KusaA1NICD和KusaONICD）来检查缺口信号。 NICD 通常抑制 Kusa-A1 和 KusaO 中成骨标记基因的表达、钙沉积和体外矿化。 Cbfa1 和 Ose2 元件的启动子活性被 NICD 减弱。 NICD 显着抑制 Kusa-A1 的体内骨形成。这些结果表明，Notch 信号传导通过细胞间相互作用充当成骨的总协调子。肾母细胞瘤过度表达基因（Nov：CCN3）是一种富含半胱氨酸的蛋白质，在禽类肾母细胞瘤中过度表达，并且被认为参与控制细胞增殖和各种组织的发育。正如最近报道 Nov 与 Notch 结合一样，我们评估了成骨间充质细胞系中的 Nov 信号传导。 Nov 表达载体转染 Kusa-A1 诱导成骨细胞标志物下调。在中央，Notch 相关基因上调。 DNA微阵列显示KusaA1NICD中Nov表达被高度诱导。因此，Nov可能诱导Notch信号传导，从而进一步诱导Nov表达，并且这个循环对于保持细胞处于不成熟状态可能是必要的。

项目成果

Differentiation of an ameloblast-lineage cell line (ALC) is induced by Sonic hedgehog signaling

• DOI: 10.1016/j.bbrc.2006.12.053
• 发表时间: 2007-02-09
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Takahashi, Satomi;Kawashima, Nobuyuki;Suda, Hideaki
• 通讯作者: Suda, Hideaki

酸化窒素合成阻害薬によるラット実験的歯髄炎におけるサイトカインおよびシクロオキシゲナーゼ2発現の制御

一氧化氮合成抑制剂对大鼠实验性牙髓炎细胞因子和环氧合酶2表达的调节

• 发表时间: 2006
• 期刊: 日本歯科保存学雑誌 49(6)
• 作者: Kawashima N.;Wongyaofa I.;Suzuki N.;Kawanishi H.;Suda H.;川島 伸之
• 通讯作者: 川島 伸之

Molecular and cell biological properties of mouse osteogenic mesenchymal progenitor cells, Kusa

• DOI: 10.1007/s00774-004-0550-y
• 发表时间: 2005-03-01
• 期刊: JOURNAL OF BONE AND MINERAL METABOLISM
• 影响因子: 3.3
• 作者: Kawashima, N;Shindo, K;Katsube, K
• 通讯作者: Katsube, K

NK cells and NKT cells in the rat dental pulp tissues.

大鼠牙髓组织中的NK细胞和NKT细胞。

• 期刊: Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontolog 2006(In press)
• 作者: Kawashima N.;Wongyaofa I.;Suzuki N.;Kawanishi H.;Suda H.
• 通讯作者: Suda H.

Effects of NOS Inhibitor on Cytokine and COX2 Expression in Rat Pulpitis

NOS抑制剂对大鼠牙髓炎细胞因子和COX2表达的影响

• 发表时间: 2006
• 期刊: The Japanese Journal of Conservative Dentistry 49(6)
• 作者: Kawashima N.;Wongyaofa I.;Suzuki N.;Kawanishi H.;Suda H.;川島 伸之;Kawashima N.
• 通讯作者: Kawashima N.