Development of ex-vivo sensitivity test for anti-cancer drugs using 3D primary culture and patient-derived xenograft

• Project/Area Number: 15K08588
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied pharmacology
• Research Institution: National Cancer Center Japan (2017-2018); Kobe University (2015-2016)
• Principal Investigator: Mukohara Toru 国立研究開発法人国立がん研究センター, 東病院, 科長 (80435718)
• Research Collaborator: Shimono Yohei
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); 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: 3次元培養 / PDX / 3D培養 / トラスツズマブ / PI3K / 脂肪幹細胞 / 初代培養 / 薬効試験

Outline of Final Research Achievements

We comparatively evaluated effects of anti-cancer drugs in 2D and 3D culture conditions using breast cancer cell lines. As a result, for cyto-toxic chemotherapeutic drugs, cells tended to be more sensitive in 2D culture than in 3D culture. On the other hand, for anti-HER2 antibody, cells are more sensitive in 3D culture. Considering efficacy in clinic, 3D culture was suggested to enhance in vivo environment.
While in 3D culture of cancer cells in malignant effusion we faced an issue of growing macrophage and mosothelial cells, we solved it by optimizing cell number to spread and serum concentration in culture media. In a project of 3D culture of patient-derived xenograft (PDX) cells, we found that co-culture with adipose-derived stem cells (ASCs) enhanced cell growth, and the effect was mediated by adipsin secreted by ASCs.

Academic Significance and Societal Importance of the Research Achievements

不確実な抗悪性腫瘍薬の効果を体外で事前に評価し、がん薬物療法の確実性を増すための技術開発は、過去数十年間種々試みられてきた。しかし、従来の2D培養では不正確であることは明らかであり、より生体に近いと推定される3D培養に期待が集まっている。我々の細胞株を用いた研究では、抗がん薬と分子標的薬でそれぞれ3D培養下での薬効が異なることを新たに見出し、3D培養に対する知見を深めることができた。当初の目標である、薬効試験に耐えうる3D培養技術の開発は達成できなかったが、体腔液内癌細胞の3D培養、脂肪幹細胞との3D共培養を試み、その礎を築くことができた。今後も、技術開発を推進したい。

Research Products

• [Journal Article] 3D Culture Represents Apoptosis Induced by Trastuzumab Better than 2D Monolayer Culture (2018)
  • Author(s): Tatara T, Mukohara T, Tanaka R, Shimono Y, Funakoshi Y, Imamura Y, Toyoda M, Kiyota N, Hirai M, Kakeji Y, Minami H.
  • Journal Title: Ａｎｔｉｃａｎｃｅｒ Ｒｅｓ Volume: 38 Issue: 5 Pages: 2831-2839
  • DOI: 10.21873/anticanres.12528
  • Peer Reviewed
• [Journal Article] Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer (2015)
  • Author(s): Imamura Y, Mukohara T, Shimono Y, Funakoshi Y, Chayahara N, Toyoda M, Kiyota N, Takao S, Kono S, Nakatsura T, Minami H
  • Journal Title: Oncol Rep Volume: 33 Issue: 4 Pages: 1837-1843
  • DOI: 10.3892/or.2015.3767
  • NAID: 120005758024
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Adipsin enhanced clonogenicity and proliferation of human breast cancer patient-derived xenograft cells. (2018)
  • Author(s): Goto H, Shimono Y, Mukohara T, et al.
  • Organizer: American Association for Cancer Research 201８
• [Presentation] 3D culture may better represent trastuzumab resistance associated with PIK3CA mutation than 2D culture (2017)
  • Author(s): Takashi Tatara, Toru Mukohara, Rina Tanaka, Yohei Shimono, Masanori Toyoda, Naomi Kiyota, Midori Hirai, Yoshihiro Kakeji, Hironobu Minami
  • Organizer: AACR annual meeting
  • Place of Presentation: Washington DC
  • Year and Date: 2017-04-01
• [Presentation] 3D culture may better represent trastuzumab resistance associated with PIK3CA mutation than 2D culture (2017)
  • Author(s): Tatara T, Mukohara T, Tanaka R, Shimono Y, Funakoshi Y, Imamura Y, Toyoda M, Kiyota N, Hirai M, Kakeji Y, Minami H.
  • Organizer: American Association for Cancer Research 2017
• [Presentation] Comparison of 2D- and 3D-cultures in in vitro evaluation of trastuzumab in HER2-overexpressing breast cancer cell lines (2016)
  • Author(s): Toru Mukohara, Rina Tanaka, Yoshinori Imamura, Tsutomu Nakagawa, Kazuhiro Yamamoto, Naoko Chayahara, Masanori Toyoda, Naomi Kiyota, Midori Hirai, Hironobu Minami
  • Organizer: 日本臨床腫瘍学会学術集会
  • Place of Presentation: 神戸市
  • Year and Date: 2016-07-28
• [Presentation] Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer (2015)
  • Author(s): Imamura Y, Mukohara T, Shimono Y, Funakoshi Y, Chayahara N, Toyoda M, Kiyota N, Takao S, Kono S, Nakatsura T, Minami H.
  • Organizer: Aｍｅｒｉｃａｎ Ａｓｓｏｃｉａｔｉｏｎ ｆｏｒ Ｃａｎｃｅｒ Ｒｅｓｅａｒｃｈ ａｎｎｕａｌ ｍｅｅｔｉｎｇ
  • Place of Presentation: Philadelphia
  • Year and Date: 2015-04-18
  • Int'l Joint Research