The elucidation of the mechanism of cellular localization of driver gene products and its medical application

• Project/Area Number: 20K21639
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 55:Surgery of the organs maintaining homeostasis and related fields
• Research Institution: National Institutes of Biomedical Innovation, Health and Nutrition
• Principal Investigator: Nishida Toshirou 国立研究開発法人医薬基盤・健康・栄養研究所, 医薬基盤研究所 細胞核輸送ダイナミクスプロジェクト, 客員研究員 (40263264)
• Co-Investigator(Kenkyū-buntansha): 小幡 裕希 国立研究開発法人国立がん研究センター, 研究所, ユニット長 (20609408) ; 黒川 量雄 国立研究開発法人理化学研究所, 光量子工学研究センター, 専任研究員 (40333504)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2021: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2020: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
• Keywords: ドライバー遺伝子 / ゴルジ体 / タンパク分解 / ユビキチン化 / プロテアソーム / タンパク代謝 / driver mutation / proteosome / lysosome / autophagosome / ドライバー遺伝子変異 / オルガネラ

Outline of Research at the Start

申請者らは、正常では細胞膜でシグナルを出すKITが、KIT変異を持つがん細胞では、低ユビキチン化状態でエンドソームやゴルジ体に停留し下流分子を恒常的に活性化し、肺癌ではEGFRの変異体はエンドソームで活性化している事を明らかにした。しかし、それら変異体が本来集積される場所以外に集積しかつ分解されない分子機構や、その分解代謝を促進する場合、どの様な経路や分解機構で代謝されるかは不明である。本研究では、超解像三次元イメージングと生化学的解析を組み合わせ、その分子機構を解明し、変異タンパクの分解誘導による新規治療戦術の開発基盤を造る。

Outline of Final Research Achievements

Mutated KIT of GIST and mutated EGFR of lung cancer are retained and activated in the Golgi apparatus and endosomes, respectively. In this study, we elucidated their retention mechanisms and explore the possibility of new development of anti-cancer agents. Mutated KIT of GIST accumulated in the Golgi apparatus is transported to lysosomes and degraded by addition of inhibitor A. Meanwhile, ubiquitination of KIT is also increased. On the contrary, treatment with HSP90 inhibitors may cause KIT degradation in proteasomes. Mutant EGFR, accumulated and activated in endosomes, was also degraded by addition of HSP90 inhibitors. Currently, we are analyzing with multicolor super-resolution three-dimensional live imaging to clarify detailed intracellular migration of molecules and kinase processing mechanism when these inhibitors are treated.

Academic Significance and Societal Importance of the Research Achievements

ドライバー遺伝子変異を持つがん細胞では、その遺伝子産物は、正常タンパク質とは異なる場所に集積し、分解を逃れ長期に増殖シグナルを出し、がん化を促進している。本研究で、GISTの変異KITのオルガネラ停留には、特定の分子が関与していること、この分子を阻害すると変異KITは選択的に本来KITが分解を受けるリソソームに輸送され分解されることが解った。一方、タンパク質の3次元構造を変えるHSP90阻害剤処理では、異常タンパク質の細胞内処理機構が働き、プロテアソームでキナーゼの分解が進むことが解った。前者は腫瘍特異的で変異キナーゼに選択的で、今後、新しい標的治療薬開発の候補となり得る。

Research Products

• [Journal Article] FLT3-ITD transduces autonomous growth signals during its biosynthetic trafficking in acute myelogenous leukemia cells (2021)
  • Author(s): Yamawaki K, Shiina I, Murata T, Tateyama S, Maekawa Y, Niwa M, Shimonaka M, Okamoto K, Suzuki T, Nishida T, Abe R, Obata Y.
  • Journal Title: Sci Rep Volume: 11 Issue: 1 Pages: 22678-22678
  • DOI: 10.1038/s41598-021-02221-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Gastrointestinal stromal tumours. (2021)
  • Author(s): Blay, JY., Kang, YK., Nishida, T. von Mehren M.
  • Journal Title: Nat Rev Dis Primers Volume: 7 Issue: 1 Pages: 22-22
  • DOI: 10.1038/s41572-021-00254-5
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Targeted therapy for drug-tolerant persister cells after imatinib treatment for gastrointestinal stromal tumours (2021)
  • Author(s): Ishida T, Takahashi T, Kurokawa Y, Nishida T, Hirota S, Serada S, Fujimoto M, Naka T, Teranishi R, Saito T, Yamashita K, Tanaka K, Yamamoto K, Makino T, Yamasaki M, Nakajima K, Eguchi H, Doki Y.
  • Journal Title: Br J Cancer Volume: 125 Issue: 13 Pages: 1511-1522
  • DOI: 10.3390/cancers13133158
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of the large‐scale Japanese patient‐derived xenograft (J‐PDX) library (2021)
  • Author(s): Yagishita Shigehiro、Kato Ken、Takahashi Mami、Imai Toshio、Yatabe Yasushi、Kuwata Takeshi、Suzuki Mikiko、Ochiai Atsushi、Ohtsu Atsushi、Shimada Kazuaki、Nishida Toshirou、Hamada Akinobu、Mano Hiroyuki
  • Journal Title: Cancer Science Volume: 112 Issue: 6 Pages: 2454-2466
  • DOI: 10.1111/cas.14899
  • Peer Reviewed / Open Access
• [Journal Article] TAS-116 inhibits oncogenic KIT signalling on the Golgi in both imatinib-naive and imatinib-resistant gastrointestinal stromal tumours. (2020)
  • Author(s): Saito Y, Takahashi T, Obata Y, Nishida T, Ohkubo S, Nakagawa F, Serada S, Fujimoto M, Ohkawara T, Nishigaki T, Sugase T, Koh M, Ishida T, Tanaka K, Miyazaki Y, Makino T, Kurokawa Y, Nakajima K, Yamasaki M, Hirota S, Naka T, Mori M, Doki Y.
  • Journal Title: Br. J. Cancer. Volume: 122 Issue: 5 Pages: 658-667
  • DOI: 10.1038/s41416-019-0688-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] MASTER KEY Project: Powering Clinical Development for Rare Cancers through a Platform Trial (2020)
  • Author(s): Okuma Hitomi S、Yonemori Kan、Narita Shoko N、Sukigara Tamie、Hirakawa Akihiro、Shimizu Toshio、Shibata Taro、Kawai Akira、Yamamoto Noboru、Nakamura Kenichi、Nishida Toshiro、Fujiwara Yasuhiro
  • Journal Title: Clinical Pharmacology & Therapeutics Volume: - Issue: 3 Pages: 596-605
  • DOI: 10.1002/cpt.1817
  • Peer Reviewed / Open Access
• [Journal Article] A New Approach to Refractory GIST with Diverse Acquired Mutations. (2020)
  • Author(s): Nishida T, Doi T.
  • Journal Title: Lancet Oncol. Volume: 21 Issue: 7 Pages: 864-865
  • DOI: 10.1016/s1470-2045(20)30209-6
  • Open Access / Int'l Joint Research
• [Presentation] 希少がん診療の現状 (2020)
  • Author(s): 西田俊朗
  • Organizer: 令和2年度 第1回大阪府がん診療連携協議会
  • Invited