High- throughput screening for personalized medicine targeting gastrointestinal cancer stem cell.

• Project/Area Number: 15K19349
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Gastroenterology
• Research Institution: Keio University
• Principal Investigator: TAKANO AI 慶應義塾大学, 医学部(信濃町), 研究員 (50647584)
• Research Collaborator: SATO Toshiro 慶應義塾大学, 医学部, 特任准教授 (70365245) ; MATANO Mami 慶應義塾大学, 医学部, 研究員 (20439889)
• Project Period (FY): 2015-04-01 – 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: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 消化器がん / HTS / 大腸癌 / がん幹細胞 / オルガノイド / PKI / スクリーニング / 幹細胞

Outline of Final Research Achievements

We have established gastrointestinal cancer organoids from the patient. As a result, we succeeded in isolating 4 kinds of tumor organoids including colorectal, gastric , liver, and pancreatic cancer from patient. It was possible to continuously and long-term culture of each of the human gastrointestinal cancer organoids by optimizing the culture condition based on the orgnoid culture protocol. We also have been successful knocked-in reporter to LGR5 locus of intestinal organoids using genome editing. Each gastrointestinal cancer organoids derived from patients became culture in 384-well plate. Furthermore, we have established tumor organoids; based High-throughput drug screen system. Image analysis was performed with 360 drugs on tumor organoids using High Contents Analyzer. Our results suggest that our established HTS for tumor organids will be apply to genetic mutation based drug discovery and personalized medicine.

Research Products

• [Journal Article] Visualization and Targeting of LGR5+ Human Colon Cancer Stem Cells (2017)
  • Author(s): Shimokawa M, Ohta Y, Nishikori S, Matano M, Takano A, Fujii M, Date S, Sugimoto S, Kanai T, Sato T
  • Journal Title: Nature Volume: 印刷中 Issue: 7653 Pages: 13-14
  • DOI: 10.1038/nature22081
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] A colorectal tumor organoid library demonstrates progressive loss of niche factor requirements. (2016)
  • Author(s): Fujii M, Shimokawa M, Date S, Takano A, Matano M, Ohta Y, Nanki K, Kawasaki K, Nakazato Y, Uraoka T, Watanabe T, Kanai T, Sato T.
  • Journal Title: Cell Stem Cell Volume: 18 Issue: 6 Pages: 827-838
  • DOI: 10.1016/j.stem.2016.04.003
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A Colorectal Tumor Organoid Library Demonstrates Progressive Loss of Niche Factor Requirements (2016)
  • Author(s): Masayuki Fujii,Mariko Shimokawa,Shoichi Date,Ai Takano, Mam iMatano,Kosaku Nanki,Yuki Ohta,Kohta Toshimitsu,YoshihiroNakazato,Kenta Kawasaki,Toshio Uraoka,Toshiaki Watanabe,Takanori Kanai,Toshiro Sato
  • Journal Title: Cell Stem Cell Volume: in press
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Modelling colorectal cancer using CRISPR-Cas9-mediated engineering of human intestinal organoids. (2015)
  • Author(s): Matano M,Date S, Shimokawa M, Takano A, Fujii M, Ohta Y, Watanabe T, Kanai T, Sato T.
  • Journal Title: Nature Medicine Volume: 21(3) Issue: 3 Pages: 256-262
  • DOI: 10.1038/nm.3802
  • Peer Reviewed / Open Access
• [Book] 細胞工学別冊 2015年上半期バイオ論文総集編 (2015)
  • Author(s): 高野愛, 佐藤俊朗
  • Total Pages: 2
  • Publisher: 学研メディカル秀潤社