Identification of phosphoproteins contributing to Epithelial-Mesenchymal Transition.

• Project/Area Number: 17K08648
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Kitasato University
• Principal Investigator: Shirakihara Takuya 北里大学, 医学部, 講師 (30548756)
• Project Period (FY): 2017-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: EMT / チロシンリン酸化 / リン酸化タンパク質 / チロシンキナーゼ / 癌 / シグナル伝達 / プロテオーム / 発生・分化

Outline of Final Research Achievements

Long-term TGF-β stimulation of epithelial cells induces myofibroblast-like changes (EMyoT), and co-stimulation of TGF-β and FGF-2 enhances EMT (eEMT). Therefore, this study aimed to reveal the phosphorylation signaling pathway involved in the induction of eEMT. Comprehensive shotgun proteomic analysis revealed approximately 250 elevated protein expression in EMyoT and eEMT, respectively, and a combined search using the SILAC method and purification with tyrosine phosphorylation antibodies revealed approximately 800 distinct protein phosphorylations in EMyoT and eEMT. Knockdown of multiple eEMT-specific phosphoproteins did not reveal any change in eEMT inducibility, but inhibitors of Axl suppressed eEMT induction.

Academic Significance and Societal Importance of the Research Achievements

TGF-βとFGF-2の長期曝露によるeEMTは創傷治癒部位や繊維化形成部位、がん微小環境など実際の生体内で誘導されている現象であることが推測できる。それ故、本研究で明らかにしたAxl阻害剤によるeEMT誘導の抑制は、EMTを標的とした治療法開発に繋がる可能性がある。また、SILAC法から新規に同定した123のeEMT特異的なチロシンリン酸化タンパク質の中にも線維化やがんの浸潤・転移促進に重要な未知のシグナル因子が含まれていることが予測される。

Research Products

• [Journal Article] Transferrin receptor 1 promotes the fibroblast growth factor receptor-mediated oncogenic potential of diffused-type gastric cancer (2022)
  • Author(s): Shirakihara Takuya、Yamaguchi Hideki、Kondo Tadashi、Yashiro Masakazu、Sakai Ryuichi
  • Journal Title: Oncogene Volume: 41 Issue: 18 Pages: 2587-2596
  • DOI: 10.1038/s41388-022-02270-5
  • Peer Reviewed
• [Presentation] FGFR2遺伝子増幅型スキルス胃がんの進展に対するTfR1の作用機序について (2022)
  • Author(s): 白木原琢哉, 堺隆一
  • Organizer: 第31回日本がん転移学会学術集会・総会
• [Presentation] The role of TfR1 in the progression of FGFR2 gene-amplified scirrhous gastric cancer (2022)
  • Author(s): 白木原琢哉, 堺隆一
  • Organizer: 第80回日本癌学会学術総会
• [Presentation] スキルス胃がんの進展に関わるFGFR2結合タンパク質の機能解析 (2021)
  • Author(s): 白木原琢哉
  • Organizer: 第30回日本がん転移学会学術集会・総会
• [Presentation] Functional analysis of FGFR2 binding target protein in scirrhous gastric cancer (2021)
  • Author(s): 白木原琢哉
  • Organizer: 第80回日本癌学会学術総会
• [Presentation] スキルス胃がんの進展に関わるFGF受容体結合タンパク質TfR1の機能解析 (2019)
  • Author(s): 白木原琢哉, 堺隆一
  • Organizer: 第28回日本がん転移学会学術集会・総会
• [Presentation] びまん性胃がんの進展に関わるFGFR2結合タンパク質の探索と機能解析 (2019)
  • Author(s): 白木原琢哉, 堺隆一
  • Organizer: 第78回日本癌学会学術総会
• [Presentation] Identification and functional analysis of FGFR2 binding proteins in diffuse-type gastric carcinoma (2019)
  • Author(s): Takuya Shirakihara, Ryuichi Sakai
  • Organizer: American Association for Cancer Research Annual Meeting
  • Int'l Joint Research
• [Presentation] The effects of FGF signaling on TGF-beta-induced EMT (2017)
  • Author(s): Takuya Shirakihara
  • Organizer: 8th International EMT Meeting
  • Int'l Joint Research