Identification of molecules that suppressed osteoclastgenesis in extracellular vesicles derived from osteosarcoma cells

• Project/Area Number: 19K18529
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 56020:Orthopedics-related
• Research Institution: Kanazawa University
• Principal Investigator: Yoshihiro Araki 金沢大学, 医学系, 協力研究員 (10800625)
• Project Period (FY): 2019-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2020: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: 骨肉腫 / 破骨細胞 / エクソソーム / マイクロRNA / 転移 / 骨微小環境の変化 / TRAF6 / miRNA146a-5p / 弾性線維関連蛋白 / NFATc1 / NFκB / IκBα / リン酸化抑制 / 血管新生 / 肺転移 / 骨肉腫由来エクソソーム

Outline of Research at the Start

骨髄環境の恒常性を維持している破骨細胞は骨肉腫細胞の存在下にその分化が抑制されて数が減少し、さらにその状況下に肺転移が促進されていることが報告されている。一方、骨肉腫など希少がんの腫瘍細胞がエクソソームの分泌を介してどのように生体内の細胞と情報伝達しているかについてはほとんど知られていない。そこで、骨肉腫細胞由来エクソソームが、骨髄環境の恒常性維持のために働いている破骨細胞にどのような分子や機序を通じてがん細胞の進展・環境形成に寄与しているかを解明することで、破骨細胞の分化を維持し、肺転移の抑制につながる骨肉腫の新たな治療戦略の可能性を探索する。

Outline of Final Research Achievements

In an orthotopic implantation study, we found that osteosarcoma-derived SEVs had the potential to enhance metastases and angiogenesis. In addition, osteosarcoma derived SEVs decreased the number of mature osteoclasts in vivo. In vitro osteoclastogenesis studies revealed that the inhibition of osteoclast maturation by osteosarcoma‐derived SEVs was mediated by suppressing the NFκB signal pathway. MicroRNA analysis of SEVs from different malignant human osteosarcomas revealed that miR-146a-5p was involved in the inhibition of osteoclastogenesis. In osteosarcoma patients, lower numbers of osteoclasts in biopsy specimens were correlated with higher malignancy. These findings indicated that osteosarcoma-derived SEVs enhance distant metastasis of osteosarcomas by inhibiting osteoclast maturation, which may be a useful prognostic marker. This diagnostic method may enable to predict malignancy at early stage, and help to provide optimal care to patients with risk of high malignancy.

Academic Significance and Societal Importance of the Research Achievements

本研究により，骨肉腫の進展に細胞外小胞が深く関与しており，その産生を抑えることで，腫瘍の浸潤・転移を抑えることができる可能性が示されました。今後，骨肉腫の早期発見や予後診断，新たな治療法の開発へと研究が発展することが期待されます。

Research Products

• [Journal Article] Osteosarcoma-Derived Small Extracellular Vesicles Enhance Tumor Metastasis and Suppress Osteoclastogenesis by miR-146a-5p (2021)
  • Author(s): Yoshihiro Araki, Hisaki Aiba, Takeshi Yoshida, Norio Yamamoto, Katsuhiro Hayashi, Akihiko Takeuchi, Shinji Miwa, Kentaro Igarashi, Tuan D. Nguyen, Kiyo-aki Ishii, Takayuki Nojima, Satoru Takahashi, Hideki Murakami, Hiroyuki Tsuchiya and Rikinari Hanayama
  • Journal Title: Frontiers in Oncology Volume: 11 Pages: 667109-667109
  • DOI: 10.3389/fonc.2021.667109
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The number of osteoclasts in a biopsy specimen can predict the efficacy of neoadjuvant chemotherapy for primary osteosarcoma (2021)
  • Author(s): Araki Yoshihiro、Yamamoto Norio、Hayashi Katsuhiro、Takeuchi Akihiko、Miwa Shinji、Igarashi Kentaro、Higuchi Takashi、Abe Kensaku、Taniguchi Yuta、Yonezawa Hirotaka、Morinaga Sei、Asano Yohei、Ikeda Hiroko、Nojima Takayuki、Tsuchiya Hiroyuki
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 1989-1989
  • DOI: 10.1038/s41598-020-80504-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structural and mechanical characteristics of exosomes from osteosarcoma cells explored by 3D-atomic force microscopy. (2021)
  • Author(s): Ayhan Yurtsever, Takeshi Yoshida, Arash Badami Behjat, Yoshihiro Araki, Rikinari Hanayama and Takeshi Fukuma
  • Journal Title: Nanoscale Volume: 13 Pages: 6661-77
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Molecular mechanism for suppression of osteoclastogenesis by osteosarcoma cell-derived extracellular vesicles (2021)
  • Author(s): Yoshihiro Araki, Norio Yamamoto, Katsuhiro Hayashi, Akihiko Takeuchi, Shinji Miwa, Kentaro Igarashi, Takashi Higuchi, Kensaku Abe, Yuta Taniguchi, Hirotaka Yonezawa, Sei Morinaga, Yohei Asano, Hiroyuki Tsuchiya
  • Organizer: 13th Asia Pacific Musculoskeletal Tumor Society
  • Int'l Joint Research
• [Presentation] 骨肉腫細胞由来エクソソームが破骨細胞の分化に及ぼす影響とその機序 (2020)
  • Author(s): 荒木麗博、山本憲男、林克洋、武内章彦、三輪真嗣、五十嵐健太郎、樋口貴史、米澤宏隆、森永整、淺野洋平、土屋弘行
  • Organizer: 第93回日本整形外科学会学術総会
• [Presentation] 骨肉腫細胞由来エクソソームが破骨細胞の分化抑制に及ぼす分子メカニズム (2019)
  • Author(s): 荒木麗博、山本憲男、林克洋、武内章彦、三輪真嗣、五十嵐健太郎、谷口裕太、米澤宏隆、森永整、淺野洋平、吉田孟史、華山力成、土屋弘行
  • Organizer: 第34回日本整形外科学会基礎学術集会
• [Presentation] 骨肉腫細胞由来エクソソームによる破骨細胞の分化抑制に関する分子メカニズム (2019)
  • Author(s): 荒木麗博、山本憲男、林克洋、武内章彦、三輪真嗣、五十嵐健太郎、谷口裕太、米澤宏隆、森永整、淺野洋平、吉田孟史、華山力成、土屋弘行
  • Organizer: 第57回日本癌治療学会
• [Patent(Industrial Property Rights)] 骨腫瘍の予後診断方法又は予後診断補助方法 (2021)
  • Inventor(s): 土屋弘行、華山力成、山本憲男、吉田孟史、荒木麗博
  • Industrial Property Rights Holder: 土屋弘行、華山力成、山本憲男、吉田孟史、荒木麗博
  • Industrial Property Rights Type: 特許
  • Filing Date: 2021