Identification of protein factors regulating secretion of specific miRNAs

• Project/Area Number: 26640095
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Tumor diagnostics
• Research Institution: National Cancer Center Japan
• Principal Investigator: Tsuchiya Naoto 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 (30322712)
• Research Collaborator: YOSHIMATSU Yuki
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 細胞外マイクロRNA / 分泌複合体 / 分泌マイクロRNA / エクソソーム / miRNA

Outline of Final Research Achievements

MicroRNA (miRNA), a class of small non-coding RNAs, have been attracting major interest in the field of cancer research to analyze molecular pathways that are deeply involved in cancer progression. In addition to their important cellular functions, secreted miRNAs (extra-cellular miRNA, ex-miRNAs) mediated by exosomes have also been considered to contribute the connection of inter-cellular networks. Although the increasing a body of evidences indicates important biological functions of ex-miRNAs, the mechanism how specific miRNAs transported into exosomes is still unknown. We, here, conducted the detailed analysis of biochemical properties for miRNA complexes, which are detectable in exosome-enriched fraction, and found that considerable number of ex-miRNAs is secreted by unknown protein complex. Furthermore, 14 proteins were identified as candidates of a novel complex, whose knockdown was suggested to regulates secretion of specific miRNAs from cancer cells.

Research Products

• [Journal Article] マイクロRNAの特性を利用した“がん医療構築戦略” (2016)
  • Author(s): 土屋直人
  • Journal Title: バイオエクススプレス Volume: 1 Pages: 8-13
• [Journal Article] An integrated prognostic classifier for stage I lung adenocarcinoma based on mRNA, microRNA and methylation biomarkers (2015)
  • Author(s): Robles AI., Kana Y., Mathe E., Okayama H., Schetter A., Brown D., Petersen D., Bowman ED., Noro R., Welsh J., Edelman D., Stevenson H., Tsuchiya N., Kohno T., Skaug V., Mollerup S., Haugen A., Meltzer P., Yokota J., Harris CC.
  • Journal Title: J Thoracic Oncol. Volume: 10 Issue: 7 Pages: 1037-1048
  • DOI: 10.1097/jto.0000000000000560
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A Three-microRNA Signature Predicts Responses to Platinum-Based Doublet Chemotherapy in Patients with Lung Adenocarcinoma. (2014)
  • Author(s): Saito M, Shiraishi K, Matsumoto K, Schetter AJ, Ogata-Kawata H, Tsuchiya N, Kunitoh H, Nokihara H, Watanabe SI, Tsuta K, Kumamoto K, Takenoshita S, Yokota J, Harris CC, Kohno T.
  • Journal Title: Clinical Cancer Research Volume: 20（18） Issue: 18 Pages: 4784-93
  • DOI: 10.1158/1078-0432.ccr-14-1096
  • Peer Reviewed / Open Access
• [Journal Article] NEK9-dependent proliferation of cancer cells lacking functional p53 (2014)
  • Author(s): Kurika D, Takeshita F, Tsuta K, Sakamoto H, Watanabe S, Matsumoto K, Watanabe M, Nakagama H, Ochiya T, Yokota J, Kohno T, Tsuchiya N.
  • Journal Title: Sci. Rep. Volume: 4 Issue: 1 Pages: 6111-6111
  • DOI: 10.1038/srep06111
  • Peer Reviewed / Open Access
• [Journal Article] Circulating Exosomal microRNAs as Biomarkers of Colon Cancer (2014)
  • Author(s): Ogata-Kawata H., Izumiya M., Kurioka D., Honnma Y., Yamada Y., Furuta K., Gunji T., Ohta H., Okamoto H., Sonoda H., Watanabe M., Nakagama H., Yokota J., Kohno T., Tsuchiya N.
  • Journal Title: Plos ONE Volume: 9 Issue: 4 Pages: e92921-e92921
  • DOI: 10.1371/journal.pone.0092921
  • Peer Reviewed / Open Access
• [Presentation] Fine-tuning of p53-dependent nucleolus stress response by miR-101 (2016)
  • Author(s): Fujiwara Y., Kohno T., Tsuchiya N.
  • Organizer: 第３９回日本分子生物学会年会
  • Place of Presentation: 神奈川県横浜市
  • Year and Date: 2016-11-30
• [Presentation] エクソソームマイクロRNAのがん診断マーカーとしての重要性 (2016)
  • Author(s): 土屋直人
  • Organizer: 第５４回日本癌治療学会学術集会
  • Place of Presentation: 神奈川県横浜市
  • Year and Date: 2016-10-20
  • Invited
• [Presentation] Nucleolar stress-specified molecular connection of miR-101 and p53 as a novel circuitry in an intrinsic tumor-suppressor network. (2016)
  • Author(s): Tsuchiya N.
  • Organizer: 第７６回日本癌学会総会
  • Place of Presentation: 神奈川県横浜市
  • Year and Date: 2016-10-06
• [Presentation] MicroRNAs in colon carcinogenesis ~ basic research to clinical applications ~ (2015)
  • Author(s): Tsuchiya N.
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 愛知県名古屋市
  • Year and Date: 2015-10-08
  • Invited
• [Presentation] Extra-cellular microRNAs - Basic research to clinical application - (2015)
  • Author(s): Tsuchiya N
  • Organizer: Seminar in Laboratory of Human Carcinogenesis, NCI.
  • Place of Presentation: Bethesda, MD, USA
  • Year and Date: 2015-04-23
  • Invited
• [Presentation] MicroRNA target screen identifies NEK9 pathway as a crucial component regulating proliferation of cancer cells lacking functional p53. (2015)
  • Author(s): Tsuchiya N.
  • Organizer: AACR Annual Meeting 2015
  • Place of Presentation: Philadelphia USA
  • Year and Date: 2015-04-17
  • Int'l Joint Research
• [Presentation] マイクロRNA：発がん機構の理解とTranslational Research (2014)
  • Author(s): 土屋 直人
  • Organizer: RNA科学総合研究センター公開シンポジウム
  • Place of Presentation: 東京理科大学
  • Year and Date: 2014-12-22
  • Invited
• [Presentation] p53不活化がん細胞のNEK9依存性増殖 (2014)
  • Author(s): 土屋 直人
  • Organizer: 第３７回分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Regulation of p53-dependent G2 arrest and apoptosis by miR-101 through the regulation of EG5 and p53 pathway (2014)
  • Author(s): Yuko Fujiwara, Takashi Kohno, Naoto Tsuchiya
  • Organizer: 第３７回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 大腸がん診断マーカーとしてのエクソソームマイクロRNAの有用性 (2014)
  • Author(s): 土屋 直人
  • Organizer: 第６９回大腸肛門病学会学術集会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-07 – 2014-11-08
  • Invited
• [Presentation] EG5-p53 axis, a novel pathway to control cell cycle and apoptosis, is regulated by miR-101 (2014)
  • Author(s): Naoto Tsuchiya
  • Organizer: 第７３回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-25 – 2015-09-27