Investigation of miRNA targeting Notch1

• Project/Area Number: 26830115
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Tumor therapeutics
• Research Institution: Tokai University
• Principal Investigator: OKUYAMA Kazuki 東海大学, 医学部, 客員准教授 (60712750)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: T-ALL / Notch1 / miRNA / Notch / T細胞性リンパ芽球性白血病 / B細胞分化

Outline of Final Research Achievements

Notch1 mutation is one of the most frequent mutation found in T-ALL. miRNA is non-coding RNA regulating target genes post-transcriptionally. Recently miRNAs are suggested as potential biologics. In this study, I investigated miRNA targeting Notch1.
Focusing on the function of Notch receptors as adhesion molecules, I established a screening system for miRNA targeting Notch1. Library of murine miRNAs was transduced into T-ALL-derived MOLT-4, and MOLT-4 cells were incubated on OP9 expressing Notch ligand, Dll4. If transduced miRNA represses Notch1 in MOLT-4, it is suggested the adhesion of MOLT4 to Dll4 would be interfered. By use of this system, I obtained 5 candidate miRNAs. One of them, miR-669m-1 functionally suppressed Notch1 in vitro. Next I performed bone-marrow transfer experiments, and found that miR-669m-1 expressing LSK cells efficiently gave rise to B cells. Since Notch1 suppresses B-lymphopoiesis, miR-669m-1 was suggested to enhance B cell development by repressing Notch1.

Research Products

• [Journal Article] 白血病細胞の分化誘導と非コードRNA miR-126 (2015)
  • Author(s): 奥山一生、幸谷愛
  • Journal Title: 感染 免疫 炎症 Volume: 45(2) Pages: 68-68
• [Journal Article] An evolutionary-conserved function of mammalian notch family members as cell adhesion molecules. (2014)
  • Author(s): Akihiko Murata, Miya Yoshino, Mari Hikosaka, Kazuki Okuyama, Lan Zhou, Seiji Sakano, Hideo Yagita, Shin-Ichi Hayashi
  • Journal Title: PLoS One Volume: 9 Issue: 9 Pages: e108535-e108535
  • DOI: 10.1371/journal.pone.0108535
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Novel functional small RNAs are selectively loaded onto mammalian Ago1 (2014)
  • Author(s): Yamakawa, N. Okuyama, K. Ogata, J. Kanai, A. Helwak, A. Takamatsu, M. Imadome, K. I. Takakura, K. Chanda, B. Kurosaki, N. Yamamoto, H. Ando, K. Matsui, H. Inaba, T. Kotani, A.
  • Journal Title: Nucleic Acids Res Volume: 未確定 Issue: 8 Pages: 5289-5301
  • DOI: 10.1093/nar/gku137
  • Peer Reviewed / Open Access
• [Journal Article] Small RNA as a regulator of hematopoietic development, immune response in infection and tumorigenesis (2014)
  • Author(s): Okuyama K, Ogata J, Yamakawa N, Chanda B, Kotani A.
  • Journal Title: International Journal of Hematology Volume: 99 Issue: 5 Pages: 553-60
  • DOI: 10.1007/s12185-014-1564-4
  • Peer Reviewed
• [Journal Article] マイクロRNAによる血液細胞分化制御 (2014)
  • Author(s): 奥山一生, 幸谷愛
  • Journal Title: 血液内科 Volume: 68 Pages: 725-729
• [Presentation] Identification of Notch-targeting miRNA that regulates B cell development and has therapeutic potential in T-ALL (2015)
  • Author(s): 那須亮、奥山一生、扇屋大輔、穂積勝人、村田暁彦、安藤 潔、幸谷愛
  • Organizer: 第74回 日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場（愛知県名古屋市）
  • Year and Date: 2015-10-08
• [Presentation] Functional and comprehensive investigation of microRNA targeting Notch receptors (2014)
  • Author(s): K. Okuyama, K. Hozumi, A. Murata, K. Ando, A. Kotani
  • Organizer: 第42回日本免疫学会総会・学術集会
  • Place of Presentation: 京都 日本
  • Year and Date: 2014-12-10
• [Presentation] The critical interaction between Epstein-Barr virus positive B-cells and tumor associated macrophages (2014)
  • Author(s): A. Sato, N. Yamakawa, K. Okuyama, A. Kotani, N. Nakamura, K. Ando
  • Organizer: 56th ASH Annual Meeting
  • Place of Presentation: CA USA
  • Year and Date: 2014-12-07
• [Presentation] EBF deficient hematopoietic progenitor cells potentially differentiated into immature B cell: EBF1 dispensable for B cell lineage commitment from pro-B to pre-B stage (2014)
  • Author(s): B. Chanda, T. Ikawa, K. Okuyama, K. Hozumi, K. Ando, A. Tojo, H. Kawamoto, A. Kotani
  • Organizer: 56th ASH Annual Meeting
  • Place of Presentation: CA USA
  • Year and Date: 2014-12-06
• [Presentation] MicroRNAライブラリーを用いたNotch受容体標的MicroRNAの探索 (2014)
  • Author(s): 奥山一生, 黒﨑なつみ, 水谷隆之, 大庭成喜, 恵口豊, 幸谷愛
  • Organizer: 第6回日本RNAi研究会
  • Place of Presentation: 広島 日本
  • Year and Date: 2014-08-29
• [Presentation] 血液腫瘍におけるMicroRNA (2014)
  • Author(s): 奥山一生, 幸谷愛
  • Organizer: 総合医学研究所・糖鎖科学研究所・創造科学技術研究機構 第6回公開合同シンポジウム
  • Place of Presentation: 神奈川 日本
  • Year and Date: 2014-07-25
  • Invited
• [Presentation] MicroRNA-126による転写因子非依存的？なB細胞運命決定制御機構 (2014)
  • Author(s): 奥山一生
  • Organizer: 第10回麒麟塾
  • Place of Presentation: 東京 日本
  • Year and Date: 2014-07-12
  • Invited