Single cell screening of molecular target in using malignant neuroblastoma animal model

• Project/Area Number: 16K10051
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Research Institute for Clinical Oncology, Saitama Cancer Center
• Principal Investigator: Takenobu Hisanori 埼玉県立がんセンター(臨床腫瘍研究所), 臨床腫瘍研究所, 研究員 (60392247)
• Co-Investigator(Kenkyū-buntansha): 上條 岳彦 埼玉県立がんセンター(臨床腫瘍研究所), 臨床腫瘍研究所, 所長 (90262708)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 幹細胞 / 発がん / 細胞分化 / トランスジェニック動物 / 小児腫瘍学

Outline of Final Research Achievements

Stem cell marker CD133 was upregulated in neuroblastoma (NB) sphere. CDX1 was identified as a transcription factor that directly induced CD133 expression in NB spheres by analysis of CD133 promoter region. CDX1 upregulated stem cell-related genes in NB cells. On the other hand, CDX1 suppressed tumor-related proliferation pathways. Then we generated CD133 and CDX1 conditional transgenic mice. These mice expressed transgenes in neural crest originated tissues.

Academic Significance and Societal Importance of the Research Achievements

神経芽腫を始めとした小児がんは、成人のがんと異なって遺伝子の変異を伴わずに発生する。したがって、発がんには何らかの遺伝子の発現異常が生じていると考えられるが、その分子機構はほとんど明らかになっていない。本研究で見出したCDX1は、神経芽腫のがん幹細胞に発現している転写因子であり、CD133をはじめとした幹細胞関連遺伝子の発現を上昇させることから、発がんだけでなく、複数の組織幹細胞の維持に関わっているものと考えられる。本課題で作出したCDX1のTgマウスは、発がんだけでなく、神経堤由来の組織の発生メカニズムを明らかにする、非常に有用なモデルとなりうる。

Research Products

• [Journal Article] EZH2 regulates neuroblastoma cell differentiation via NTRK1 promoter epigenetic modifications (2018)
  • Author(s): Li Zhenghao、Takenobu Hisanori、Setyawati Amallia Nuggetsiana、Akita Nobuhiro、Haruta Masayuki、Satoh Shunpei、Shinno Yoshitaka、Chikaraishi Koji、Mukae Kyosuke、Akter Jesmin、Sugino Ryuichi P.、Nakazawa Atsuko、Nakagawara Akira、Aburatani Hiroyuki、Ohira Miki、Kamijo Takehiko
  • Journal Title: Oncogene Volume: 37 Issue: 20 Pages: 2714-2727
  • DOI: 10.1038/s41388-018-0133-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] CFC1 is a cancer stemness-regulating factor in neuroblastoma (2017)
  • Author(s): Chikaraishi Koji、Takenobu Hisanori、Sugino Ryuichi P.、Mukae Kyosuke、Akter Jesmin、Haruta Masayuki、Kurosumi Masafumi、Endo Takaho A.、Koseki Haruhiko、Shimojo Naoki、Ohira Miki、Kamijo Takehiko
  • Journal Title: Oncotarget Volume: 8 Issue: 28 Pages: 45046-45059
  • DOI: 10.18632/oncotarget.18464
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Reevaluation of PDGFRα signal in neuroblastoma: high expression and a new nonsynonymous mutation (2018)
  • Author(s): Shunpei Satoh, Miki Ohira, Hisanori Takenobu, Akira Nakagawara, Takehiko Kamijo
  • Organizer: Advanced Neuroblastoma Research 2018
  • Int'l Joint Research
• [Presentation] CDX1 regulates neuroblastoma stemness through MYC pathway modulation and reprogramming gene activation (2018)
  • Author(s): Hisanori Takenobu, Miki Ohira, Ryuichi Sugino, Koji Chikaraishi, Kyosuke Mukae, Nobuhiro Akita, Masayuki Haruta, Akira Nakagawara, Manabu Nakayama, Haruhiko Koseki, Takehiko Kamijo
  • Organizer: Advanced Neuroblastoma Research 2018
  • Int'l Joint Research
• [Presentation] EZH2 regulates neuroblastoma cell differentiation via NTRK1 promoter epigenetic modifications (2018)
  • Author(s): Zhenghao Li, Hisanori Takenobu, Amallia Nuggetsiana Setyawati, Nobuhiro Akita, Masayuki Haruta, Shunpei Satoh, Yoshitaka Shinno, Koji Chikaraishi, Kyosuke Mukae, Jesmin Akter, Ryuichi P. Sugino, Atsuko Nakazawa, Akira Nakagawara, Hiroyuki Aburatani, Miki Ohira, Takehiko Kamijo
  • Organizer: Advanced Neuroblastoma Research 2018
  • Int'l Joint Research
• [Presentation] CFC1 is a cancer stemness-regulating factor in neuroblastoma. (2018)
  • Author(s): Chikaraishi K, Takenobu H, Sugino RP, Mukae K, Akter J, Haruta M, Kurosumi M, Endo TA, Koseki H, Shimojo N, Ohira M, Kamijo T
  • Organizer: Advanced Neuroblastoma Research 2018
  • Int'l Joint Research
• [Presentation] Identification of molecules that regulate the sensitivity of EZH2 inhibitor in neuroblastoma (2018)
  • Author(s): Yuki Endo, Hisanori Takenobu, Ryuichi Sugino, Masayuki Haruta, Kyosuke Mukae, Shunpei Satoh, Yoshitaka Shinno, Mariko Hasegawa, Ryu Okada, Yutaka Katai, Masaki Nio, Miki Ohira, Takehiko Kamijo
  • Organizer: 第77回日本癌学会学術総会
• [Presentation] Polycomb molecule L3MBTL2 suppresses apoptotic cell death cooperating with BMI1 in neuroblastoma (2018)
  • Author(s): Ryu Okada, Hisanori Takenobu, Miki Ohira, Ryuichi Sugino, Koji Chikaraishi, Nobuhiro Akita, Masayuki Haruta, Kyosuke Mukae, Yuki Endo, Shunpei Satoh, Yutaka Katai, Haruhiko Koseki, Takehiko Kamijo
  • Organizer: 第77回日本癌学会学術総会
• [Presentation] CDX1 regulates stemness through MYC pathway modulation and reprogramming gene activation in neuroblastoma (2018)
  • Author(s): Hisanori Takenobu, Miki Ohira, Ryuichi Sugino, Koji Chikaraishi, Ryu Okada, Kyosuke Mukae, Yuki Endo, Parvin Sultana, Yutaka Katai, Nobuhiro Akita, Masayuki Haruta, Haruhiko Koseki, Takehiko Kamijo
  • Organizer: 第77回日本癌学会学術総会
• [Presentation] Development of neuroblastoma model from iPS-based neural crest cells for analysis of neuroblastoma tumorigenesis (2018)
  • Author(s): Kyosuke Mukae, Hisanori Takenobu, Ryuichi Sugino, Miki Ohira, Shunpei Satoh, Yuki Endo, Ryu Okada, Masayuki Haruta, Junya Toguchida, Kenji Osafune, Tatsutoshi Nakahata, Takehiko Kamijo
  • Organizer: 第77回日本癌学会学術総会
• [Presentation] EZH2 regulates neuroblastoma cell differentiation via NTRK1 promoter epigenetic modifications (2018)
  • Author(s): Zhenghao Li, Hisanori Takenobu, Amallia Nuggetsiana Setyawati, Nobuhiro Akita, Masayuki Haruta, Shunpei Satoh, Yoshitaka Shinno, Koji Chikaraishi, Kyosuke Mukae, Jesmin Akter, Ryuichi P. Sugino, Atsuko Nakazawa, Akira Nakagawara, Hiroyuki Aburatani, Miki Ohira, Takehiko Kamijo
  • Organizer: 第60回 日本小児血液がん学会
• [Presentation] Screening of EZH2i effector molecules in neuroblastoma by multi-omics analysis (2018)
  • Author(s): Yuki Endo, Hisanori Takenobu, Ryuichi Sugino, Masayuki Haruta, Kyosuke Mukae, Shunpei Satoh, Yoshitaka Shinno, Mariko Hasegawa, Ryu Okada, Yutaka Katai, Masaki Nio, Miki Ohira, Takehiko Kamijo
  • Organizer: 第60回 日本小児血液がん学会
• [Presentation] Effect of ATRX deletion by CRISPR-Cas9 system in neuroblastoma (2018)
  • Author(s): Yutaka Katai, Jesmin Akter, Hisanori Takenobu, Miki Ohira, Ryuichi Sugino, Masayuki Haruta, Shunpei Satoh, Kyosuke Mukae, Parvin Sultana, Takehiko Kamijo
  • Organizer: 第41回 日本分子生物学会
• [Presentation] Neuroblastoma cell dedifferentiation via epigenetic modification (2018)
  • Author(s): Hisanori Takenobu, Ryuichi P. Sugino, Miki Ohira, Takehiko Kamijo
  • Organizer: 第41回 日本分子生物学会
  • Invited
• [Presentation] CFC1 regulates cancer stemness phenotypes in neuroblastoma via Activin signaling suppression (2017)
  • Author(s): Koji Chikaraishi, Hisanori Takenobu, Ryuichi Sugino, Kyosuke Mukae, Masayuki Haruta, Masafumi Kurosumi, Yoshitaka Shinno, Akter Jesmin, A Takaho Endo, Haruhiko Koseki, Miki Ohira, Takehiko Kamijo
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] Sphere specific transcription factor CDX1 has dual functions for regulating stemness of neuroblastoma (2017)
  • Author(s): Hisanori Takenobu, Ryuichi Sugino, Miki Ohira, Koji Chikaraishi, Kyosuke Mukae, Masayuki Haruta, Haruhiko Koseki, Takehiko Kamijo
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] Characterization of cell type by DNA methylation pattern using principal component analysis-based approach (2017)
  • Author(s): Ryuichi Sugino, Miki Ohira, Satoshi Yamashita, Hisanori Takenobu, Junya Toguchida, Toshikazu Ushijima, Takehiko Kamijo
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] EZH2 regulates neuroblastoma cell differentiation via NTRK1 promoter epigenetic modifications (2017)
  • Author(s): Zhenghao Li, Hisanori Takenobu, Masayuki Haruta, Shunpei Satoh, Yoshitaka Shinno, Kyosuke Mukae, Jesmin Akter, Ryuichi Sugino, Akira Nakagawara, Hiroyuki Aburatani, Miki Ohira, Takehiko Kamijo
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] CRISPR/Cas9-based knockout of the chromatin remodeling factor ATRX and its effect in neuroblastoma (NB) cells (2017)
  • Author(s): Jesmin Akter, Yutaka Katai, Hisanori Takenobu, Masayuki Haruta, Parvin Sultana, Kyosuke Mukae, Ryuichi Sugino, Miki Ohira, Takehiko Kamijo
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] EZH2 regulates neuroblastoma cell differentiation via NTRK1 promoter epigenetic modifications (2017)
  • Author(s): Hisanori Takenobu, Zhenghao Li, Masayuki Haruta, Shunpei Satoh, Yoshitaka Shinno, Kyosuke Mukae, Jesmin Akter, Ryuichi Sugino, Akira Nakagawara, Hiroyuki Aburatani, Miki Ohira, Takehiko Kamijo
  • Organizer: The 3rd Asia-Pacific International Symposium of Neuroblastoma
• [Presentation] EZH2 regulates neuroblastoma cell differentiation via NTRK1 promoter epigenetic modifications (2017)
  • Author(s): Hisanori Takenobu, Zhenghao Li, Amallia Nuggetsiana Setyawati, Nobuhiro Akita, Masayuki Haruta, Shunpei Satoh, Yoshitaka Shinno, Koji Chikaraishi, Kyosuke Mukae, Jesmin Akter, Yuki Endo, Ryuichi P. Sugino, Akira Nakagawara, Hiroyuki Aburatani, Miki Ohira, Takehiko Kamijo
  • Organizer: 第59回 日本小児血液がん学会学術集会
• [Presentation] Sphere specific transcription factor CDX1 regulates stemness of neuroblastoma (2017)
  • Author(s): Hisanori Takenobu, Ryuichi Sugino, Miki Ohira, Koji Chikaraishi, Kyosuke Mukae, Yuki Endo, Sultana Parvin, Ryu Okada, Yutaka Katai, Masayuki Haruta, Haruhiko Koseki, Takehiko Kamijo
  • Organizer: 第59回 日本小児血液がん学会学術集会
• [Presentation] Effect of CFC1 on cancer cell stemness in neuroblastoma (2017)
  • Author(s): Koji Chikaraishi, Hisanori Takenobu, Ryuichi Sugino, Kyosuke Mukae, Jesmin Akter, Masayuki Haruta, Masafumi Kurosumi, Takaho Endo, Haruhiko Koseki, Naoki Shimojo, Miki Ohira, Takehiko Kamijo
  • Organizer: 第59回 日本小児血液がん学会学術集会
• [Presentation] ヒトiPS細胞を用いた神経芽腫発がんモデル (2016)
  • Author(s): Takenobu H, Sugino R, Ohira M, Yamashita S, Toguchida J, Kamijo T
  • Organizer: 第58回日本小児血液がん学会学術集会
  • Place of Presentation: 品川
  • Year and Date: 2016-12-15
• [Presentation] CFC-1遺伝子は神経芽腫細胞の幹細胞化制御因子である (2016)
  • Author(s): Chikaraishi K, Takenobu H, Mukae K, Sugino R, Ohira M, Koseki H, Kamijo T
  • Organizer: 第58回日本小児血液がん学会学術集会
  • Place of Presentation: 品川
  • Year and Date: 2016-12-15
• [Presentation] 神経芽腫幹細胞を制御する新規シグナル経路の解析 (2016)
  • Author(s): Hisanori Takenobu, Miki Ohira, Koji Chikaraishi, Kyosuke Mukae, Nobuhiro Akita, Masayuki Haruta, Haruhiko Koseki, Takehiko Kamijo
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-06
• [Presentation] iPS細胞から神経堤細胞への分化における統合解析 (2016)
  • Author(s): Sugino R, Ohira M, Yamashita S, Takenobu H, Toguchida J, Ushijima T, Kamijo T
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-06
• [Presentation] EZH2による神経芽腫の分化抑制 (2016)
  • Author(s): Li Z, Setyawati AN, Takenobu H, Akita N, Haruta M, Chikaraishi K, Mukae K, Akter J, Sugino R, Ohira M, Kamijo K
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-06
• [Presentation] CDX1 regulates cancer stemness pathway in neuroblastoma, CDX1 regulates cancer stemness pathway in neuroblastoma (2016)
  • Author(s): Hisanori Takenobu, Miki Ohira, Koji Chikaraishi, Nobuhiro Akita, Yohko Yamaguchi, Masayuki Haruta, Akira Nakagawara, Manabu Nakayama, Haruhiko Koseki, Takehiko Kamijo
  • Organizer: 国際神経芽腫学会 (ANR2016)
  • Place of Presentation: ケアンズ (オーストラリア)
  • Year and Date: 2016-06-19
  • Int'l Joint Research