Identification of molecular targets for liquid biopsy in patients with bone and soft tissue sarcoma

• Project/Area Number: 16H05449
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Okayama University
• Principal Investigator: Ozaki Toshifumi 岡山大学, 医歯薬学総合研究科, 教授 (40294459)
• Co-Investigator(Kenkyū-buntansha): 杉原 進介 独立行政法人国立病院機構四国がんセンター(臨床研究センター), その他部局等, 医長 (60314671) ; 清野 正普 岡山大学, 大学病院, 医員 (60756097) ; 國定 俊之 岡山大学, 医歯薬学総合研究科, 准教授 (80346428) ; 横尾 賢 岡山大学, 大学病院, 医員 (00781204) ; 森田 卓也 岡山大学, 大学病院, 医員 (10743007) ; 藤原 智洋 岡山大学, 医歯薬学総合研究科, 助教 (80639211) ; 魚谷 弘二 岡山大学, 大学病院, 医員 (30708087)
• Research Collaborator: OCHIYA Takahiro ; YOSHIOKA Yusuke ; UEDA Koji ; UEHARA Takenori
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2018: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2017: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
• Keywords: liquid biopsy / 骨軟部肉腫 / バイオマーカー / エクソソーム / Liquid biopsy / exosome / Ewing 肉腫

Outline of Final Research Achievements

Lack of circulating biomarker is a crucial problem for bone and soft tissue sarcomas. To improve prognosis in patients, identification of sensitive circulating biomarkers is important, since current diagnostic modalities have limitations in monitoring tumor or drug sensitivity during/after treatments for bone and soft tissue sarcomas. In our study, we identified the circulating exosomal marker by proteomic analysis using Ewing sarcoma cell culture media and serum samples from Ewing sarcoma patients. In patients with osteosarcoma, we identified serum miR-25-3p as a circulating biomarker that is secreted from OS cells. Serum miR-25-3p levels reflected tumor response to chemotherapy and recurrence, and were significantly correlated with prognosis in patients. We also identified serum miR-92b-3p as a diagnostic target of liquid biopsy for synovial sarcoma. Overall, our data indicated promising diagnostic role of circulating miRNAs and exosomes for bone and soft tissue sarcomas.

Academic Significance and Societal Importance of the Research Achievements

骨軟部肉腫は、全身の骨や軟部組織から発生するまれな悪性腫瘍である。近年、患者体液を用いた体液診断技術が悪性腫瘍の新しい診療ツールとして開発されつつある。本研究では、骨軟部肉腫循血中腫瘍由来exosomeや分泌型miRNAが特定され、治療経過や臨床病理学的な関連性が見出された。これらのマーカーが、化学療法抵抗性や転移の早期発見の新たな診断技術になれば、肉腫患者予後の改善に寄与することが期待できる。

Research Products

• [Journal Article] Clinical and Functional Significance of Intracellular and Extracellular microRNA-25-3p in Osteosarcoma (2018)
  • Author(s): Yoshida A, Fujiwara T, Uotani K, Morita T, Kiyono M, Yokoo S, Hasei J, Nakata E, Kunisada T, Ozaki T.
  • Journal Title: Acta Medica Okayama Volume: 72 Issue: 2 Pages: 165-174
  • DOI: 10.18926/AMO/55857
  • NAID: 120006459963
  • URL: https://ousar.lib.okayama-u.ac.jp/55857
  • Peer Reviewed / Open Access
• [Journal Article] Clinical significance of circulating miR-25-3p as a novel diagnostic and prognostic biomarker in osteosarcoma (2017)
  • Author(s): Fujiwara T, Uotani K, Yoshida A, Morita T, Nezu Y, Kobayashi E, Yoshida A5, Uehara T1, Omori T1, Sugiu K1, Komatsubara T, Takeda K, Kunisada T, Kawamura M, Kawai A, Ochiya T, Ozaki T
  • Journal Title: Oncotarget Volume: - Issue: 20 Pages: 33375-33392
  • DOI: 10.18632/oncotarget.16498
  • Peer Reviewed / Open Access
• [Journal Article] Circulating MicroRNA-92b-3p as a Novel Biomarker for Monitoring of Synovial Sarcoma (2017)
  • Author(s): Uotani Koji、Fujiwara Tomohiro、Yoshida Aki、Iwata Shintaro、Morita Takuya、Kiyono Masahiro、Yokoo Suguru、Kunisada Toshiyuki、Takeda Ken、Hasei Joe、Numoto Kunihiko、Nezu Yutaka、Yonemoto Tsukasa、Ishii Takeshi、Kawai Akira、Ochiya Takahiro、Ozaki Toshifumi
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 14634-14634
  • DOI: 10.1038/s41598-017-12660-5
  • Peer Reviewed / Open Access
• [Presentation] The role of exosomes in osteosarcoma cell migration, invasion and lung metastasis (2019)
  • Author(s): Masahiro Kiyono; Tomohiro Fujiwara; Aki Yoshida; Suguru Yokoo; Koji Demiya; Yusuke Mochizuki; Joe Hasei; Eiji Nakata; Toshiyuki Kunisada; Toshiyuki Ozaki
  • Organizer: ORS 2019 Annual Meeting
  • Int'l Joint Research
• [Presentation] Clinical and functional significance of single circulating and cellular microRNA dysregulation in osteosarcoma (2019)
  • Author(s): Aki Yoshida; Tomohiro Fujiwara; Koji Uotani; Takuya Morita; Masahiro Kiyono; Suguru Yokoo; Joe Hasei; Eiji Nakata; Toshiyuki Kunisada; Toshifumi Ozaki
  • Organizer: ORS 2019 Annual Meeting
  • Int'l Joint Research
• [Presentation] Detection of exosomal markers in patients with synovial sarcoma (2019)
  • Author(s): Suguru Yokoo; Tomohiro Fujiwara; Koji Uotani; Aki Yoshida; Takuya Morita; Masahiro Kiyono; Toshiyuki Kunisada; Yusuke Yoshioka; Koji Ueda; Takahiro Ochiya; Toshifumi Ozaki
  • Organizer: ORS 2019 Annual Meeting
  • Int'l Joint Research
• [Presentation] 滑膜肉腫由来exosomeの特定とマーカーの網羅的解析 (2018)
  • Author(s): 横尾 賢, 藤原 智洋, 吉田 晶, 清野 正普, 望月 雄介, 出宮 光二, 長谷井 嬢, 国定 俊之, 吉岡 祐亮, 植田 幸嗣, 落谷 孝広, 尾崎 敏文
  • Organizer: 第77回日本癌学会総会
• [Presentation] 骨肉腫における細胞内外の単一microRNAの機能について (2018)
  • Author(s): 吉田 晶, 藤原 智洋, 魚谷 弘二, 森田 卓也, 清野 正普, 横尾 賢, 長谷井 嬢, 国定 俊之, 尾崎 敏文
  • Organizer: 第77回日本癌学会総会
• [Presentation] Identification of surface marker protein by proteomic profiling of exosomes derived from Ewing sarcoma cells (2017)
  • Author(s): Aki Yoshida, Tomohiro Fujiwara, Koji Uotani, Yusuke Yoshioka, Koji Ueda, Takuya Morita, Masahiro Kiyono, Kazuhisa Sugiu, Tadashi Komatsubara, Jo Hasei, Toshiyuki Kunisada, Takahiro Ochiya, Toshifumi Ozaki
  • Organizer: Orthopaedic research Society 2017 Annual Meeting
  • Place of Presentation: San Diego, USA
  • Year and Date: 2017-03-20
  • Int'l Joint Research
• [Presentation] 骨肉腫細胞内と分泌エクソソーム内におけるmicroRNA発現プロファイリングの相違と患者循環エクソソーム内の発現評価 (2017)
  • Author(s): 清野 正普, 藤原 智洋, 魚谷 弘二, 吉田 晶, 森田 卓也, 杉生 和久, 小松原 将, 望月 雄介, 長谷井 嬢, 国定 俊之, 尾崎 敏文
  • Organizer: 第50回日本整形外科学会骨・軟部腫瘍学術集会
• [Presentation] 悪性骨・軟部腫瘍転移制御への道 粘液線維肉腫における分泌型microRNAは腫瘍の浸潤性と関連しているか (2017)
  • Author(s): 森田 卓也, 藤原 智洋, 魚谷 弘二, 吉田 晶, 清野 正普, 杉生 和久, 小松原 将, 望月 雄介, 長谷井 嬢, 国定 俊之, 尾崎 敏文
  • Organizer: 第50回日本整形外科学会骨・軟部腫瘍学術集会
• [Presentation] Ewing肉腫における腫瘍由来exosomeを標的としたliquid biopsy法の確立 (2017)
  • Author(s): 魚谷 弘二, 藤原 智洋, 吉田 晶, 岩田 慎太郎, 森田 卓也, 清野 正普, 長谷井 嬢, 国定 俊之, 吉岡 祐亮, 植田 幸嗣, 落谷 孝広, 尾崎 敏文
  • Organizer: 第50回日本整形外科学会骨・軟部腫瘍学術集会
• [Presentation] Liquid Biopsy in Patients with Sarcoma (2016)
  • Author(s): Toshifumi Ozaki
  • Organizer: The 60th Anniversary Congress of the Korean Orthopaedic Association & The 61st Regular General Assembly
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2016-10-19
  • Int'l Joint Research
• [Presentation] Ewing肉腫から分泌されるexosomeの表面蛋白を利用したliquid biopsyの開発 (2016)
  • Author(s): 望月雄介，魚谷弘二，藤原智洋，吉田晶，森田卓也，清野正普，国定俊之，吉岡裕亮，植田幸嗣，落谷孝広，尾崎敏文
  • Organizer: 第31回日本整形外科学会基礎学術集会
  • Place of Presentation: 福岡市
  • Year and Date: 2016-10-13