Development of novel mouse brain tumor model using in vivo electroporation and piggyBac system

• Project/Area Number: 16K07125
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Tumor biology
• Research Institution: Keio University
• Principal Investigator: Onishi Nobuyuki 慶應義塾大学, 医学部(信濃町), 訪問研究員 (40534540)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); 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: マウス脳腫瘍モデル / In vivoエレクトロポレーション / piggyBacシステム / 神経幹細胞

Outline of Final Research Achievements

Glioblastoma multiforme (GBM), is one of the most malignant brain tumors, has highly-proliferative and invasive characters. To understand these malignant characters of GBM, an appropriate carcinogenesis model is required. Loss of INK4A/ARF and stimulation of common signal transduction pathways involving RAS are frequently found in GBM. Previously, we have established a stable mouse models of brain tumors, transplanting the genetically modified neural stem cells (NSCs). Recently, for the purpose of reproducing a clinical tumor initiating process, we are developing a novel mouse model of brain tumors by gene transfer into mouse brain directly. Using in vivo electroporation and piggyBac system, transduction of activated-H-RAS and shInk4a/Arf into NSCs in mouse brain, efficiently formed highly proliferative, invasive and heterogeneous brain tumors. Based on these findings, we propose this in vivo carcinogenesis technique is efficient method to generate appropriate mouse brain tumor models.

Academic Significance and Societal Importance of the Research Achievements

悪性脳腫瘍、特にグリオブラストーマ(glioblastoma multiforme: GBM)は原発性脳腫瘍のうち悪性度が最も高く、浸潤の早さから手術による全摘は困難とされており、平均生存期間は約1年と極めて予後不良な悪性腫瘍である。GBMは放射線・化学療法に抵抗性を持ち、効果的な治療は未だ確立されていない。GBMの性状を理解し、新たな治療戦略を考案するためには適切な発がんモデルの構築が必須である。本研究で構築したマウス脳腫瘍モデルは、ヒトGBMに酷似した特徴を有する脳腫瘍を簡便な方法で作製することができ、最も予後が悪く難治性のがんの一つであるGBMの治療を目指した研究を促進するものである。

Research Products

• [Journal Article] Development of a functional thyroid model based on an organoid culture system. (2018)
  • Author(s): Saito Y, Onishi N, Takami H, Seishima R, Inoue H, Hirata Y, Kameyama K, Tsuchihashi K, Sugihara E, Uchino S, Ito K, Kawakubo H, Takeuchi H, Kitagawa Y, Saya H, Nagano O.
  • Journal Title: Biochem Biophys Res Commun Volume: 497 Issue: 2 Pages: 783-789
  • DOI: 10.1016/j.bbrc.2018.02.154
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Activation of Transforming Growth Factor Beta 1 Signaling in Gastric Cancer-associated Fibroblasts Increases Their Motility, via Expression of Rhomboid 5 Homolog 2, and Ability to Induce Invasiveness of Gastric Cancer Cells. (2017)
  • Author(s): Ishimoto T, Miyake K, Nandi T, Yashiro M, Onishi N, Huang KK, Joyce LN, Kalpana R, Tay ST, Suzuki Y, Cho BC, Kuroda D, Arima K, Izumi D, Iwatsuki M, Baba Y, Oki E, Watanabe M, Saya H, Hirakawa K, Baba H, Tan P.
  • Journal Title: Gastroenterology Volume: in press Issue: 1 Pages: 191-204
  • DOI: 10.1053/j.gastro.2017.03.046
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Simvastatin-Induced Apoptosis in Osteosarcoma Cells: A Key Role of RhoA-AMPK/p38 MAPK Signaling in Antitumor Activity. (2017)
  • Author(s): Kamel WA, Sugihara E, Nobusue H, Yamaguchi-Iwai S, Onishi N, Maki K, Fukuchi Y, Matsuo K, Muto A, Saya H, Shimizu T.
  • Journal Title: Molecular Cancer Therapeutics Volume: 16 Issue: 1 Pages: 182-192
  • DOI: 10.1158/1535-7163.mct-16-0499
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The EGF receptor promotes the malignant potential of glioma by regulating amino acid transport system xc(-) (2016)
  • Author(s): Tsuchihashi K, Okazaki S, Ohmura M, Ishikawa M, Sampetrean O, Onishi N, Wakimoto H, Yoshikawa M, Seishima R, Iwasaki Y, Morikawa T, Abe S, Takao A, Shimizu M, Masuko T, Nagane M, Furnari FB, Akiyama T, Suematsu M, Baba E, Akashi K, Saya H, Nagano O.
  • Journal Title: Cancer Research Volume: in press Issue: 10 Pages: 2954-2963
  • DOI: 10.1158/0008-5472.can-15-2121
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Tumour resistance in induced pluripotent stem cells derived from naked mole-rats (2016)
  • Author(s): Miyawaki S, Kawamura Y, Oiwa Y, Shimizu A, Hachiya T, Bono H, Koya I, Okada Y, Kimura T, Yoshihiro Tsuchiya, Suzuki S, Onishi N, Kuzumaki N, Matsuzaki Y, Narita M, Ikeda E, Okanoya K, Seino K, Saya H, Okano H & Miura K
  • Journal Title: Nature Communications Volume: 7 Issue: 1 Pages: 11471-11471
  • DOI: 10.1038/ncomms11471
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] In vivoエレクトロポレーションを用いたマウス発がんモデルならびに簡便な遺伝子改変マウス作製法の開発 (2018)
  • Author(s): 大西 伸幸、大塚 正人、佐藤 正宏、佐谷 秀行
  • Organizer: 第77回 日本癌学会学術総会
• [Presentation] Development of novel mouse brain tumor model using in vivo electroporation and piggyBac system (2016)
  • Author(s): Nobuyuki Onishi, Hideyuki Saya
  • Organizer: 第75回 日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜（神奈川）
  • Year and Date: 2016-10-06
• [Presentation] In vivoエレクトロポレーションを用いた新規マウス脳腫瘍モデルの開発 (2016)
  • Author(s): 大西 伸幸
  • Organizer: 平成28年度新学術領域研究「学術研究支援基盤形成【先端モデル動物支援プラットフォーム】」若手支援技術講習会
  • Place of Presentation: 蓼科グランドホテル滝の湯（長野）
  • Year and Date: 2016-09-14
• [Presentation] In vivoエレクトロポレーションを用いた新規マウス脳腫瘍モデルの開発ならびに性状解析 (2016)
  • Author(s): 大西 伸幸、佐谷 秀行
  • Organizer: 第4回 がんと代謝研究会
  • Place of Presentation: かごしま県民交流センター（鹿児島）
  • Year and Date: 2016-07-07
• [Presentation] In vivoエレクトロポレーションを用いた新規マウス脳腫瘍モデルの開発 (2016)
  • Author(s): 大西 伸幸、佐谷 秀行
  • Organizer: 第43回 BMSコンファレンス (BMS2016)
  • Place of Presentation: ホテル ニューアカオ（静岡）
  • Year and Date: 2016-07-04
• [Presentation] Development of mouse brain tumor model using in vivo electroporation and piggyBac system (2016)
  • Author(s): Nobuyuki Onishi, Hideyuki Saya
  • Organizer: 第14回 幹細胞シンポジウム
  • Place of Presentation: 淡路夢舞台国際会議場（兵庫）
  • Year and Date: 2016-05-20