Challenge to elucidate molecular pathology of sudden death by focusing on the unique phenomenon that "blood does not clot"

• Project/Area Number: 17K19905
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Health science and related fields
• Research Institution: Osaka University
• Principal Investigator: Nagano Kazuya 大阪大学, 薬学研究科, 准教授 (40548301)
• Project Period (FY): 2017-06-30 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2019: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2017: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 突然死 / プロスタノイド

Outline of Final Research Achievements

In this study, a simultaneous quantification system for prostanoids (PNs) tried to be constructed, to understand the alternation of PN expression profiles which may be related to of the phenomenon "blood does not clot" unique to sudden death. In quantifying PNs, stable metabolites in urine were focused, because PNs were unstable in blood. By purification using a reverse phase column, the matrix effects against the final PGD2, PGE2, and PGF2α metabolites were low, and their accuracies were nearly 100%. In contrast, the matrix effects against the final PGI2 and TXA2 metabolites were high. By combining reverse phase and ion exchange columns, the matrix effects decreased so that the accuracy was nearly 100%. To validate the method, the urinary metabolites of Crohn’s disease (CD) patients and healthy individuals were quantified. The data showed that final PGE2 metabolite levels were significantly higher in CD patients, so that the urinary metabolite profiles of CD patients is determined.

Academic Significance and Societal Importance of the Research Achievements

プロスタノイド：PNは、生理機能を担っている一方で、その破綻は、病態の発症/悪化に関連することが知られている。特に、痛覚過敏では、PGE2とPGI2が相互に協調し合って働いている。そのため、生体内のPN変動を理解することが、突然死を始め、各種病態解明に繋がると期待される。しかし、PNは血中半減期が短いため、適切な定量は困難であった。
本研究では、上記課題を解決するため、二段階固相抽出法を応用し、独自の尿中PN代謝物一斉定量系を開発した。また、ヒト尿試料を用い、その実用性を示してきた。今後、突然死を始め、様々な疾患に本手法を応用することで、新たな分子病態の解明と、診断/治療への応用が期待される。

Research Products

• [Journal Article] Development and evaluation of a simultaneous and efficient quantification strategy for final prostanoid metabolites in urine (2019)
  • Author(s): Tian-qi Zhang, Hirotaka Kuroda, Kazuya Nagano, Soshi Terada, Jian-Qing Gao, Kazuo Harada, Kazumasa Hirata, Hirofumi Tsujino, Kazuma Higashisaka, Hiroshi Matsumoto, Yasuo Tsutsumi
  • Journal Title: Prostaglandins, Leukotrienes and Essential Fatty Acids Volume: 6 Pages: 102032-102032
  • DOI: 10.1016/j.plefa.2019.102032
  • Peer Reviewed / Int'l Joint Research
• [Presentation] 乳がん遠隔転移症例における尿中プロスタノイド代謝物量の解析 (2020)
  • Author(s): 張 天斉, 長野一也, 黒田博隆, 寺田壮志, 原田和生, 平田收正, 辻野博文, 東阪和馬, 松本博志, 堤 康央
  • Organizer: 日本薬学会第140年会
• [Presentation] プロスタノイドの尿中最終代謝物同時定量法の最適化と応用 (2019)
  • Author(s): 張 天斉, 長野一也, 黒田博隆, 寺田壮志, 原田和生, 平田收正, 辻野博文, 東阪和馬, 松本博志, 堤 康央
  • Organizer: 日本薬学会第139年会
• [Presentation] Development and evaluation of a simultaneous quantification method for final metabolites of prostanoids in urine. (2018)
  • Author(s): Zhang T., Nagano K., Kuroda H., Terada S., Harada K., Hirata K., Tsujino H., Higashisaka K., Matsumoto H., Tsutsumi Y.
  • Organizer: 第12回次世代を担う若手医療薬科学シンポジウム
• [Presentation] プロスタノイドの尿中最終代謝物同時定量法の確立とその評価 (2018)
  • Author(s): 張 天斉, 長野一也, 黒田博隆, 原田和生, 平田收正, 東阪和馬, 松本博志, 堤 康央
  • Organizer: 日本薬学会第138年会