Development of our novel method to analyze cancer-derived circulating exosomes

• Project/Area Number: 16K11002
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Urology
• Research Institution: Osaka University
• Principal Investigator: Ujike Takeshi 大阪大学, 医学系研究科, 助教 (00738875)
• Co-Investigator(Kenkyū-buntansha): 野々村 祝夫 大阪大学, 医学系研究科, 教授 (30263263) ; 中田 渡 大阪大学, 医学系研究科, 招へい教員 (30648019) ; 植村 元秀 大阪大学, 医学系研究科, 特任准教授(常勤) (40631015)
• Research Collaborator: TSUJIKAWA kazutake ; JINGUSHI kentaro ; UEDA koji
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 腎がん / エクソソーム / 血液バイオマーカー / 腎細胞癌 / タンパク発現解析 / 癌 / タンパク

Outline of Final Research Achievements

Cancer-associated extracellular vesicles (EVs) are intimately involved in establishment of tumor microenvironment and the occurrence of metastasis. Hence we extracted EVs directly from surgically resected viable clear cell renal cell carcinoma (ccRCC) tissues and adjacent normal renal tissues. We obtained EVs directly from fresh clear cell renal cell carcinoma (ccRCC) clinical specimens by an ultracentrifuge method. Obtained tissue-exudative EVs (Te-EVs) were analyzed by LC/MS/MS analysis. Quantitative LC/MS analysis identified 3,871 tissue-exudative EV (Te-EV) proteins, among which 106 proteins were highly enriched in tumor Te-EVs. Importantly, several cancer-specific ptoteins were also significantly higher in serum EVs from ccRCC patients compared to those from healthy donors. In summary, this is the first experimental record reporting extraction of EVs from viable human tissue samples (Te-EVs) and identification of a novel serum biomarker.

Academic Significance and Societal Importance of the Research Achievements

本研究において開発した手法は特許申請（バイオマーカー,疾患関連遺伝子の探索方法、及び腎がんマーカー：特願2018-547766）を行った。またその方法を用いて探索同定したタンパクは血液中にがん患者特異的に検出され、血液バイオマーカーとしての臨床応用を目指して研究を継続する予定である。

Research Products

• [Journal Article] Leukocyteassociated immunoglobulinlike receptor 1 promotes tumorigenesis in RCC (2019)
  • Author(s): Jingushi, K., Uemura, M., Nakano, K., Hayashi, Y., Wang, C., Ishizuya, Y., Yamamoto, Y., Hayashi, T., Kinouchi, T., Matsuzaki, K., Kato, T., Kawashima, A., Ujike, T., Nagahara, A., Fujita, K., Ueda, K., Tsujikawa, K., and Nonomura, N.
  • Journal Title: Oncol Rep Volume: 41 Pages: 1293-1303
  • DOI: 10.3892/or.2018.6875
  • Peer Reviewed / Open Access