Analysis of genes related to the ectopic bone formation induced by mechanical stress and search for drugs targeting the genes.

分析与机械应力诱导的异位骨形成相关的基因并寻找针对该基因的药物。

• 批准号: 16390063
• 负责人: FURUKAWA Ken-Ichi
• 金额: $ 7.74万
• 依托单位: Hirosaki 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 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390063/
• 关键词: ectopic bone formation; mechanical stress; microarray; gene expression; siRNA; Cbfa1; nucleotide receptor; endochondral ossification

To clarify the cause of the ossification of posterior longitudinal ligament (OPLL), comprehensive analyses by cDNA microarray method were carried out using cultured human spinal ligament cells that had been subjected to uniaxial cyclic stretching (mechanical stress). Microarray analyses showed the up-regulation of many ossification-marker genes including alkaline phosphatase, osteocalcin, collagen type I and Cbfa1, by cyclic stretching in ligament cells from OPLL patients but not in normal ligament cells, suggesting that OPLL cells are higher sensitive to mechanical stress than normal ligament cells. The higher sensitivity to mechanical stress was also confirmed in ligament tissue from OPLL patients. These results suggest that mechanical stress participates in the progression of OPLL through changes in expressions of various genes related to bone metabolism.Spectrum of P2 receptor subtype expression in OPLL cells was quite different from that in non-OPLL cells. Especially, P2Y1 expression in OPLL cells was much higher than that in non-OPLL cells and almost equal to that in SaOS2 (osteosarcoma cell line). ATP as a P2Y1 agonist increased the expressions of osteogenic markers such as ALP and osteopontin in OPLL cells but not in non-OPLL cells. These increases were almost diminished in the presence of P2Y1-specific antagonist. Furthermore, mechanical stress increased ATP release from OPLL cells remarkably. These results suggest that as one of extracellular factors influencing the process of ossification, a pathophysiological action of ATP via P2Y1 is important. They become a novel pharmacotherapeutic target for OPLL.

为了阐明后纵韧带骨化（OPLL）的原因，采用基因芯片技术对体外培养的人脊柱韧带细胞进行了单轴周期性拉伸（机械应力）后的OPLL进行了综合分析。微阵列分析显示，上调的许多骨化标志物基因，包括碱性磷酸酶，骨钙素，I型胶原和Cbfa1，周期性拉伸韧带细胞从OPLL患者，但不是在正常的韧带细胞，表明OPLL细胞比正常的韧带细胞更敏感的机械应力。在OPLL患者的韧带组织中也证实了对机械应力的更高敏感性。这些结果表明，机械应力通过改变骨代谢相关基因的表达参与了OPLL的发生发展，OPLL细胞中P2受体亚型的表达谱与非OPLL细胞有很大不同。尤其是P2Y1在OPLL细胞中的表达明显高于非OPLL细胞，与骨肉瘤细胞SaOS 2的表达基本一致。ATP作为P2Y1激动剂可增加OPLL细胞中成骨标志物如ALP和骨桥蛋白的表达，但对非OPLL细胞无影响。在P2Y1特异性拮抗剂的存在下，这些增加几乎减少。此外，机械应力显著增加OPLL细胞ATP释放。这些结果表明，作为影响骨化过程的细胞外因素之一，ATP通过P2Y1的病理生理作用是重要的。它们成为OPLL的一个新的药理学靶点。

项目成果

Molecular basis of ectopic bone formation induced by mechanical stress.

机械应力诱导异位骨形成的分子基础。

• 发表时间: 2006
• 期刊: Journal of Pharmacological Science 100
• 作者: Yamaya M.;Shibahara S.;Ken-Ichi Furukawa;Ken-Ichi Furukawa
• 通讯作者: Ken-Ichi Furukawa

メカニカルストレスと生体応答

机械应力和生物反应

• 发表时间: 2004
• 期刊: 化学と生物 42
• 作者: Ituro Inoue;et al.;Ken-Ichi Furukawa;Ken-Ichi Furukawa;Ryuji Ikeda;Ryuji Ikeda et al.;Koei Iwasaki;Koei Iwasaki et al.;Koei Iwasaki;古川賢一
• 通讯作者: 古川賢一

脊柱靱帯骨化症発症におけるメカニカルストレスの関与

机械应力参与脊柱韧带骨化病的发生

• 发表时间: 2004
• 期刊: 別冊整形外科 45
• 作者: Ituro Inoue;et al.;Ken-Ichi Furukawa;Ken-Ichi Furukawa;Ryuji Ikeda;Ryuji Ikeda et al.;Koei Iwasaki;Koei Iwasaki et al.;Koei Iwasaki;古川賢一;岩崎弘英
• 通讯作者: 岩崎弘英

難病と在宅ケア

疑难杂症及家庭护理

• 发表时间: 2006
• 期刊: 脊柱靱帯骨化症の薬物治療法開発を目指して 12
• 作者: Yamaya M.;Shibahara S.;Ken-Ichi Furukawa;Ken-Ichi Furukawa;Ituro Inoue;古川賢一
• 通讯作者: 古川賢一

Molecular basis of cctopic bone formation induced by mechanical stress.

机械应力诱导异位骨形成的分子基础。

• 发表时间: 2006
• 期刊: Journal of Pharmacological Science 100
• 作者: Ituro Inoue;et al.;Ken-Ichi Furukawa;Ken-Ichi Furukawa
• 通讯作者: Ken-Ichi Furukawa