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 P2Y1 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 P2Y1。分别通过实时定量聚合酶链反应和免疫荧光染色研究mRNA和蛋白质的表达。转染后，P2Y1 瞬时转染 2 天后，与非 OPLL 患者的细胞相比，OPLL 患者来源的脊柱韧带细胞中的骨形态发生蛋白 2 (BMP-2) 和 Sox9 mRNA 表达显着增加。免疫荧光分析显示，BMP-2和P2Y1表达仅在OPLL患者中增加，而Sox9表达在OPLL和非OPLL患者中增加。 MRS2279 是一种选择性 P2Y1 拮抗剂，在 P2Y1 强制表达后阻断 Sox9 和 BMP-2 的上调。此外，短暂转染 P2Y1 4 天后，仅在 OPLL 患者的脊髓韧带细胞中观察到矿化。这些结果表明P2Y1表达在OPLL患者脊柱韧带异位骨形成中起重要作用，调节P2Y1功能的药物将成为治疗OPLL的潜在候选药物。

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

Calcification of aortic smooth muscle cells isolated from spontaneously hypertensive rats.

• DOI: 10.1254/jphs.fp0072013
• 发表时间: 2008
• 期刊: Journal of pharmacological sciences
• 影响因子: 3.5
• 作者: K. Kanemaru;K. Seya;I. Miki;S. Motomura;K. Furukawa

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

根据颈椎后纵韧带骨化分类的成骨分化能力的遗传差异。

• 发表时间: 2010
• 作者: Kudo H;Ono A;Numasawa T;Wada W;Tanaka S;Asari1 T;Toh S;Motomura S;Furukawa K.-I.
• 通讯作者: Furukawa K.-I.

TNF-αはNF-kB経路を介して石灰化大動脈弁狭窄症患者由来の大動脈弁間質細胞の石灰化を促進する

TNF-α通过NF-kB途径促进钙化主动脉瓣狭窄患者的主动脉瓣间质细胞钙化

• 发表时间: 2010
• 作者: 杉浦寧;三村喬生;望月大二郎;徳野博信;臼井節夫;田中いく子;本多芳子;児玉亨;石橋英俊;中村俊;小柴満美子;于在強,瀬谷和彦,大徳和之,元村成,福田幾夫,古川賢一
• 通讯作者: 于在強,瀬谷和彦,大徳和之,元村成,福田幾夫,古川賢一

Genetic Association in Progression of the Ossification Area of the Posterior Longitudinal Ligament of the Cervical Spine

颈椎后纵韧带骨化区进展的遗传关联

• 发表时间: 2009
• 作者: Dong YF;Liu L;Kataoka K;Nakamura T;Fukuda M;Tokutomi Y;et al.;Furukawa Ken-Ichi
• 通讯作者: Furukawa Ken-Ichi