Exploring molecular targets for the establishment of drug therapy for the ossification of the posterior longitudinal ligament of the spine

探索建立脊柱后纵韧带骨化药物治疗的分子靶点

• 批准号: 20590245
• 负责人: FURUKAWA Kenichi
• 金额: $ 3.08万
• 依托单位: Hirosaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20590245/
• 关键词: 異所性骨化; 脊柱靱帯; 機械的ストレス; 薬物治療; 脊柱靱帯骨化症; 細胞外ATP受容体; 幹細胞; gremlin; BMP2; P2Y1; 過剰発現; 骨化様式; TSG-6; 形質転換; アルカリフォスファターゼ; メカニカルストレス; 細胞外ヌクレオチド受容体; 遺伝子導入; ATP

To verify the hypothesis that P2Y1 expression causes ossification in the spinal ligaments, we forced expression of P2Y1 in spinal ligament cells obtained from OPLL and non-OPLL patients. The expression of mRNA and protein was investigated by quantitative real-time polymerase chain reaction and immunofluorescence staining, respectively. After transfection, bone morphogenetic protein-2 (BMP-2) and Sox9 mRNA expression was significantly increased in spinal ligament cells derived from OPLL patients compared with cells from non-OPLL patients 2 days after P2Y1 transient transfection. Immunofluorescence analysis showed that BMP-2 and P2Y1 expression was increased in OPLL patients only, while Sox9 expression was increased in OPLL and non-OPLL patients. MRS2279, a selective P2Y1 antagonist, blocked the upregulation of Sox9 and BMP-2 after forced expression of P2Y1. Furthermore, 4 days after transient transfection of P2Y1, mineralization was observed only in spinal ligament cells from OPLL patients. These results suggest that P2Y1 expression plays an important role in ectopic bone formation in the spinal ligaments of OPLL patients and that a drug which modulates the P2Y1 function will be a potential candidate for the drug in the therapy of OPLL.

为了验证P2 Y1表达导致脊柱韧带骨化的假设，我们在从OPLL和非OPLL患者获得的脊柱韧带细胞中强制表达P2 Y1。采用实时定量聚合酶链反应（RT-PCR）和免疫荧光染色法检测mRNA和蛋白的表达。转染后，骨形态发生蛋白-2（BMP-2）和Sox 9 mRNA的表达显着增加脊髓韧带细胞来源于OPLL患者相比，非OPLL患者的细胞P2 Y1瞬时转染后2天。免疫荧光分析显示BMP-2和P2 Y1仅在OPLL患者中表达增加，而Sox 9在OPLL和非OPLL患者中表达增加。MRS 2279是一种选择性P2 Y1拮抗剂，可阻断P2 Y1强制表达后Sox 9和BMP-2的上调。此外，瞬时转染P2 Y1后4天，仅在OPLL患者的脊柱韧带细胞中观察到矿化。这些结果表明，P2 Y1表达在OPLL患者脊柱韧带异位骨形成中起重要作用，并且调节P2 Y1功能的药物将是OPLL治疗药物的潜在候选者。

项目成果

高血圧によるラット大動脈平滑筋の異所性石灰化における一酸化窒素の役割

一氧化氮在高血压诱导的大鼠主动脉平滑肌异位钙化中的作用

• 发表时间: 2010
• 作者: Cui W;Matsuno K;Iwata K;Ibi M;Katsuyama M;Kakehi T;Sasaki M;Ikami K;Zhu K;Yabe-Nishimura C.;古川賢一
• 通讯作者: 古川賢一

Genetic Differences in the Osteogenic Differentiation Potency According to the Classification of Ossification of the Posterior Longitudinal Ligament of the Cervical Spine

• DOI: 10.1097/brs.0b013e3181e9a8a6
• 发表时间: 2011-05-20
• 期刊: SPINE
• 影响因子: 3
• 作者: Kudo, Hitoshi;Furukawa, Ken-Ichi;Toh, Satoshi
• 通讯作者: Toh, Satoshi

Genetic differences in the osteogenic differentiation potency based onaccording to the classification for the ossification of the posterior longitudinal ligament of the cervical spine.

成骨分化能力的遗传差异基于颈椎后纵韧带骨化的分类。

• 发表时间: 2010
• 期刊: Spine (印刷中)
• 作者: Kudo H;Furukawa K-I;他11名
• 通讯作者: 他11名

A functional RNAi scree, n for runx2-regulated genes corresponding to ectopic bone formation in human spinal ligaments

功能性 RNAi 筛选，用于与人类脊柱韧带异位骨形成相对应的 runx2 调节基因

• 发表时间: 2008
• 期刊: Journal of Pharmacological Sciences 106
• 作者: 田中進;児玉亨;本多芳子;臼井節夫;本多真;Kishiya Masaki
• 通讯作者: Kishiya Masaki

脊椎後縦靭帯骨化症患者におけるP2Y1受容体の病態生理学的役割一過剰発現による解析

P2Y1受体在后纵韧带骨化患者中的病理生理作用——过表达分析

• 发表时间: 2009
• 作者: Dong YF;Liu L;et al;古川賢一
• 通讯作者: 古川賢一