Establishment of Analysis of Genetic Diversity and Mutations and Method for Diagnosing Malignancy of Salivary Gland Tumor using Salivary Exosomes

• Project/Area Number: 16K11524
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pathobiological dentistry/Dental radiology
• Research Institution: Tokyo Dental College
• Principal Investigator: Kokubun Katsutoshi 東京歯科大学, 歯学部, 講師 (90535808)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: エクソソーム / バイオマーカー / DNA / 腫瘍 / exosome / 唾液 / DNA解析

Outline of Final Research Achievements

This study explored the nature of dsDNA contained in the exosomes of cancer cells. Ultracentrifugation was used to purify the exosomes, and the expression of the exosome markers was confirmed by FACS and Western blot analysis. DNA was extracted from the exosomes, and the size of the DNA was evaluated by Bioanalyzer. TEM observation confirmed the expression of dsDNA in the exosomes. Analysis of dsDNA in the exosome revealed that it reflects a tumor specific gene mutation possessed by the parent cell. In addition, whole exome sequencing indicated that the DNA of the parent cell matches the DNA in the exosomes.

Academic Significance and Societal Importance of the Research Achievements

本研究により、癌細胞由来のエクソソームを解析することで、癌細胞の遺伝子情報を得ることができ、診断やその後の治療方針に大変有効な情報を得ることができることがわかった。これにより、血液や唾液といった患者の体液中のエクソソームを解析するだけで、癌の遺伝子情報を踏まえた診断が可能となることも期待できる。

Research Products

• [Journal Article] Platelets Enhance Multiple Myeloma Progression via IL-1β Upregulation (2018)
  • Author(s): Takagi S, Tsukamoto S, Park J, Johnson KE, Kawano Y, Moschetta M, Liu CJ, Mishima Y, Kokubun K, Manier S, Salem KZ, Huynh D, Sacco A, Forward J, Roccaro AM, Battinelli EM, Ghobrial IM.
  • Journal Title: Clinical Cancer Research Volume: 24 Issue: 10 Pages: 2430-2439
  • DOI: 10.1158/1078-0432.ccr-17-2003
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Blocking IFNAR1 inhibits multiple myeloma?driven Treg expansion and immunosuppression (2018)
  • Author(s): Kawano Y, Zavidij O, Park J, Moschetta M, Kokubun K, Mouhieddine TH, Manier S, Mishima Y, Murakami N, Bustoros M, Pistofidis RS, Reidy M, Shen YJ, Rahmat M, Lukyanchykov P, Karreci ES, Tsukamoto S, Shi J, Takagi S, Huynh D, Sacco A, Tai YT, Chesi M, Bergsagel PL, Roccaro AM, Azzi J, Ghobrial IM.
  • Journal Title: Journal of Clinical Investigation Volume: 128 Issue: 6 Pages: 2487-2499
  • DOI: 10.1172/jci88169
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Inhibition of microRNA-138 enhances bone formation in multiple myeloma bone marrow niche (2018)
  • Author(s): Tsukamoto S, Lovendorf MB, Park J, Salem KZ, Reagan MR, Manier S, Zavidij O, Rahmat M, Huynh D, Takagi S, Kawano Y, Kokubun K, Thrue CA, Nagano K, Petri A, Roccaro AM, Capelletti M, Baron R, Kauppinen S, Ghobrial IM.
  • Journal Title: Leukemia Volume: 32 Issue: 8 Pages: 1739-1750
  • DOI: 10.1038/s41375-018-0161-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Congenital Epulis: A Case and Review of the Literature (2018)
  • Author(s): Kokubun Katsutoshi、Matsuzaka Kenichi、Akashi Yoshihiko、Sumi Masami、Nakajima Kei、Murakami Satoshi、Narita Masato、Shibahara Takahiko、Inoue Takashi
  • Journal Title: The Bulletin of Tokyo Dental College Volume: 59 Issue: 2 Pages: 127-132
  • DOI: 10.2209/tdcpublication.2017-0028
  • NAID: 130007386303
  • ISSN: 0040-8891
  • Peer Reviewed / Open Access
• [Journal Article] Profiling of circulating exosomal miRNAs in patients with Waldenstr?m Macroglobulinemia (2018)
  • Author(s): Bouyssou JM, Liu CJ, Bustoros M, Sklavenitis-Pistofidis R, Aljawai Y, Manier S, Yosef A, Sacco A, Kokubun K, Tsukamoto S, Perilla Glen A, Huynh D, Castillo JJ, Treon SP, Leblond V, Hermine O, Roccaro AM, Ghobrial IM, Capelletti M.
  • Journal Title: PLOS ONE Volume: 13 Issue: 10 Pages: e0204589-e0204589
  • DOI: 10.1371/journal.pone.0204589
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Immobilization of a carbon nanomaterial-based localized drug-release system using a bispecific material-binding peptide. (2018)
  • Author(s): Kokubun K, Matsumura S, Yudasaka M, Iijima S, Shiba K
  • Journal Title: Int J Nanomedicine. Volume: 13 Pages: 1643-1652
  • DOI: 10.2147/ijn.s155913
  • Peer Reviewed / Open Access
• [Presentation] 上顎洞に発生した ALK 陽性未分化大細胞リンパ腫の一例 (2018)
  • Author(s): 中條 貴俊, 明石 良彦, 根本 淳, 小谷地雅秀、中島 啓, 國分, 克寿, 橋本 和彦, 菅原 圭亮, 松坂 賢一, 井上 孝
  • Organizer: 第２９回日本臨床口腔病理学会・第１１回日本口腔検査学会総会共催学術大会
• [Presentation] 顎骨内のリンパ増殖性病変が診断の契機となったIgG4関連疾患の一例 (2018)
  • Author(s): 中島 啓, 中條 貴俊, 鷲見 正美, 明石 良彦, 國分 克寿, 橋本 和彦, 佐々木 文, 冨田 茂樹, 松坂 賢一, 井上 孝
  • Organizer: 第107回日本病理学会
• [Presentation] ラット唾液腺創傷部に入れたマトリゲルの影響 in vivoからin vitroでの検討 (2018)
  • Author(s): 金内 洋光, 小林 史卓, 遠藤 多加史, 外村 岳央, 明石 良彦, 鷲見 正美, 根本 淳, 中條 貴俊, 矢野 尚, 中島 啓, 國分 克寿, 村上 聡, 松坂 賢一, 井上 孝
  • Organizer: 第305回東京歯科大学学会
• [Presentation] FGF7はラット唾液腺創傷治癒における幹細胞の細胞増殖と分化を促進させる (2018)
  • Author(s): 遠藤 多加史, 小林 史卓, 金内 洋光, 外村 岳央, 明石 良彦, 鷲見 正美, 根本 淳, 中條 貴俊, 中島 啓, 矢野 尚, 國分 克寿, 村上 聡, 松坂 賢一, 井上 孝
  • Organizer: 第305回東京歯科大学学会