Establishment of diffuse astrocytoma cell line utilizing advanced culture system

• Project/Area Number: 16K15641
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Neurosurgery
• Research Institution: Kumamoto University (2017-2018); The University of Tokyo (2016)
• Principal Investigator: 武笠 晃丈 熊本大学, 大学院生命科学研究部(医), 教授 (90463869)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); 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: 神経膠腫 / 星細胞腫 / IDH遺伝子 / 細胞モデル / 星細胞種

Outline of Annual Research Achievements

本研究は、従来型培養方法で樹立困難なIDH遺伝子変異神経膠腫の細胞モデルを、種々の培養方法の工夫を用いることで作成することを目的とした。当初は、正常アストロサイトに変異遺伝子などの遺伝子導入を試みるも、星細胞腫モデルとしてより適しているATRX遺伝子の不活化した細胞の作成は困難であった。そこで、種々の悪性化過程にあるIDH変異星細胞腫腫瘍検体の初代培養を試みたが、継代を続けると細胞増殖が困難となり、やはり樹立にいたらなかった。
そこで平成30年度の繰越期間にむけ、初代培養細胞を一度iPS細胞化し、その後にこれを増殖させ、モデル細胞として用いることとした。まず、その前段階として、正常アストロサイトをiPS化する条件確立のため、エピソーマルベクターにてiPS細胞誘導を行ったところ、7系統のiPS細胞様の細胞株樹立に成功した。次に、IDH変異星細胞腫のiPS化技術確立のため、まずは、悪性度の高い膠芽腫の初代培養により作成した膠芽腫幹細胞様株からのiPS細胞作成を行うこととした。そのため、膠芽腫臨床切除検体を用いて、継続的な膠芽腫幹細胞様株の樹立を行った。これらの膠芽腫幹細胞様株に関しては、xenograft としての腫瘍形成能などの評価を順次施行した。そして、これら膠芽腫幹細胞様株を用い、アストロサイト同様のエピソーマルベクターを用いた方法を基盤として、様々な条件を検討しつつiPS細胞株化を図ったが、残念ながら研究期間中には神経膠腫からのiPS細胞樹立には至らなかった。さらなる技術的改良が必要と考えられた。また、神経膠腫細胞株の増殖を促進する培養条件を検討するため、種々の基質を用いた3次元及び2次元培養法の比較検討による評価を行ったところ、細胞株により増殖を促進する基質があることが明らかとなり、これらを使い分けることによりIDH変異星細胞株の樹立が可能となる可能性も示唆された。

Research Products

• [Journal Article] Pattern of FDG and MET Distribution in High- and Low-Grade Gliomas on PET Images. (2019)
  • Author(s): Takahashi Miwako、Soma Tsutomu、Mukasa Akitake、Tanaka Shota、Yanagisawa Shunsuke、Momose Toshimitsu
  • Journal Title: Clinical Nuclear Medicine Volume: 44 Pages: 265-271
  • DOI: 10.1097/rlu.0000000000002460
  • Peer Reviewed
• [Journal Article] DNA demethylation is associated with malignant progression of lower-grade gliomas. (2019)
  • Author(s): Nomura Masashi、Saito Kuniaki、Aihara Koki、Nagae Genta、Yamamoto Shogo、Tatsuno Kenji、Ueda Hiroki、Fukuda Shiro、Umeda Takayoshi、Tanaka Shota、…、Narita Yoshitaka、Nagane Motoo、Nishikawa Ryo、Ueki Keisuke、Saito Nobuhito、Aburatani Hiroyuki、Mukasa Akitake
  • Journal Title: Sci Rep Volume: 9 Pages: 1903-1903
  • DOI: 10.1038/s41598-019-38510-0
  • Peer Reviewed / Open Access
• [Journal Article] Oligodendrocyte Progenitor Cells and Macrophages/Microglia Produce Glioma Stem Cell Niches at the Tumor Border (2018)
  • Author(s): Hide T, Komohara Y, Miyasato Y, Nakamura H, Makino K, Takeya M, Kuratsu JI, Mukasa A, Yano S.
  • Journal Title: EBioMedicine Volume: 30 Pages: 94-104
  • DOI: 10.1016/j.ebiom.2018.02.024
  • Peer Reviewed / Open Access
• [Journal Article] Employment status and termination among survivors of pediatric brain tumors: a cross-sectional survey (2018)
  • Author(s): Sato I, Higuchi A, Yanagisawa T, Murayama S, Kumabe T, Sugiyama K, Mukasa A, Saito N, Sawamura Y, Terasaki M, Shibui S, Takahashi J, Nishikawa R, Ishida Y, Kamibeppu K.
  • Journal Title: International Journal of Clinical Oncology Volume: － Pages: 801-811
  • DOI: 10.1007/s10147-018-1279-2
  • Peer Reviewed
• [Journal Article] IDH-mutated astrocytomas with 19q-loss constitute a subgroup that confers better prognosis. (2018)
  • Author(s): Otani Ryohei、Uzuka Takeo、Higuchi Fumi、Matsuda Hadzki、Nomura Masashi、Tanaka Shota、Mukasa Akitake、Ichimura Koichi、Kim Phyo、Ueki Keisuke
  • Journal Title: Cancer Sci Volume: 109 Pages: 2327-2335
  • DOI: 10.1111/cas.13635
  • Peer Reviewed / Open Access
• [Journal Article] Concomitant administration of radiation with eribulin improves the survival of mice harboring intracerebral glioblastoma. (2018)
  • Author(s): Miki S, Imamichi S, Fujimori H, Tomiyama A, Fujimoto K, Satomi K, Matsushita Y, Matsuzaki S, Takahashi M, Ishikawa E, Yamamoto T, Matsumura A, Mukasa A, Nishikawa R, Masutomi K, Narita Y, Masutani M, Ichimura K.
  • Journal Title: Cancer Sci Volume: 109 Pages: 2275-2285
  • DOI: 10.1111/cas.13637
  • NAID: 120006501402
  • Peer Reviewed / Open Access
• [Journal Article] BCL2 expression is associated with a poor prognosis independent of cellular origin in primary central nervous system diffuse large B-cell lymphoma. (2018)
  • Author(s): Makino K, Nakamura H, Shinojima N, Kuroda JI, Yano S, Mikami Y, Mukasa A
  • Journal Title: J Neurooncol Volume: 140 Pages: 115-121
  • DOI: 10.1007/s11060-018-2940-3
  • Peer Reviewed / Open Access
• [Journal Article] SIRT2-mediated inactivation of p73 is required for glioblastoma tumorigenicity. (2018)
  • Author(s): Funato K, Hayashi T, Echizen K, Negishi L, Shimizu N, Koyama-Nasu R, Nasu-Nishimura Y, Morishita Y, Tabar V, Todo T, Ino Y, Mukasa A, Saito N & Akiyama T.
  • Journal Title: EMBO Rep Volume: 19
  • DOI: 10.15252/embr.201745587
  • Peer Reviewed / Open Access
• [Journal Article] Machine learning based on multi-parametric magnetic resonance imaging to differentiate glioblastoma multiforme from primary cerebral nervous system lymphoma. (2018)
  • Author(s): Nakagawa M, Nakaura T, Namimoto T, Kitajima M, Uetani H, Tateishi M, Oda S, Utsunomiya D, Makino K, Nakamura H, Mukasa A, Hirai T, Yamashita Y
  • Journal Title: Eur J Radiol Volume: 108 Pages: 147-154
  • DOI: 10.1016/j.ejrad.2018.09.017
  • Peer Reviewed / Open Access
• [Journal Article] CSF TACI and BAFF levels in patients with primary CNS lymphoma as novel diagnostic biomarkers (2018)
  • Author(s): Mizutani H, Nakane S, Ikeda T, Nakamura H, Takamatsu K, Makino K, Tawara N, Mukaino A, Watari M, Matsui H, Mukasa A, Ando Y
  • Journal Title: Ann Clin Transl Neurol. Volume: 5 Pages: 1611-1616
  • DOI: 10.1002/acn3.668
  • Peer Reviewed / Open Access
• [Journal Article] 脳腫瘍のゲノム診療に向けて (2018)
  • Author(s): 武笠 晃丈
  • Journal Title: Clinical Neuroscience Volume: 36 Pages: 539-542
• [Journal Article] Identification of a novel fusion gene HMGA2-EGFR in glioblastoma. (2018)
  • Author(s): Komuro Akiyoshi、Raja Erna、Iwata Caname、Soda Manabu、Isogaya Kazunobu、Yuki Keiko、Ino Yasushi、Morikawa Masato、Todo Tomoki、Aburatani Hiroyuki、Suzuki Hiromichi、Ranjit Melissa、Natsume Atsushi、Mukasa Akitake、Saito Nobuhito、Okada Hitoshi、Mano Hiroyuki、Miyazono Kohei、Koinuma Daizo
  • Journal Title: Int J Cancer. Volume: 142 Pages: 1627-1639
  • DOI: 10.1002/ijc.31179
  • Peer Reviewed
• [Journal Article] Clinical significance of polyglutamylation in primary central nervous system lymphoma. (2018)
  • Author(s): Shinojima Naoki、Fujimoto Kenji、Makino Keishi、Todaka Kohei、Yamada Kazumichi、Mikami Yoshiki、Oda Kazutaka、Nakamura Kazumi、Jono Hirofumi、Kuratsu Jun-ichi、Nakamura Hideo、Yano Shigetoshi、Mukasa Akitake
  • Journal Title: Acta Neuropathol Commun. Volume: 6 Pages: 15-15
  • DOI: 10.1186/s40478-018-0522-4
  • Peer Reviewed / Open Access
• [Journal Article] 神経膠腫の不均一性による治療抵抗性とその治療戦略 (2018)
  • Author(s): 武笠 晃丈
  • Journal Title: 実験医学 Volume: 36 Pages: 307-313
• [Journal Article] Genome-wide DNA methylation profiling identifies primary central nervous system lymphoma as a distinct entity different from systemic diffuse large B-cell lymphoma (2017)
  • Author(s): Nakamura T, Yamashita S, Fukumura K, Tanaka K, Nagane M, Narita Y, Mano H, Ushijima T, Ichimura K
  • Journal Title: Acta Neuropathologica Volume: 133 Pages: 321-324
  • DOI: 10.1007/s00401-016-1664-8
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] An automated voxel-based method for calculating the reference value for a brain tumour metabolic index using 18F-FDG-PET and 11C-methionine PET (2017)
  • Author(s): Takahashi, Soma T, Mukasa A, Koyama, Arai T, Momose T.
  • Journal Title: Annals of Nuclear Medicine Volume: 31 Pages: 250-259
  • DOI: 10.1007/s12149-017-1153-8
  • Peer Reviewed / Open Access
• [Journal Article] Differences in genetic and epigenetic alterations between von Hippel?Lindau disease?related and sporadic hemangioblastomas of the central nervous system (2017)
  • Author(s): Takayanagi S, Mukasa A, Tanaka S, Nomura M, Omata M, Yanagisawa S, Yamamoto S, Ichimura K, Nakatomi H, Ueki K, Aburatani H, Saito N
  • Journal Title: Neuro-Oncology Volume: 印刷中 Pages: 1228-1236
  • DOI: 10.1093/neuonc/nox034
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Brachyury gene copy number gain and activation of the PI3K/Akt pathway: association with upregulation of oncogenic Brachyury expression in skull base chordoma (2017)
  • Author(s): Otani Ryohei、Mukasa Akitake、Shin Masahiro、Omata Mayu、Takayanagi Shunsaku、Tanaka Shota、Ueki Keisuke、Saito Nobuhito
  • Journal Title: Journal of Neurosurgery Volume: なし Pages: 1-10
  • DOI: 10.3171/2016.12.jns161444
  • Peer Reviewed
• [Journal Article] Distinct molecular profile of diffuse cerebellar gliomas (2017)
  • Author(s): Nomura M, Mukasa A, Nagae G, Yamamoto S, Tatsuno K, Ueda H, Fukuda S, Umeda T, Suzuki T, Otani R, Kobayashi K, Maruyama T, Tanaka S, Takayanagi S, Nejo T, Takahashi S, Ichimura K, Nakamura T, Muragaki Y, Narita Y, Nagane M, Ueki K, Nishikawa R, Shibahara J, Aburatani H, Saito N
  • Journal Title: Acta Neuropathol. Volume: 134 Pages: 941-956
  • DOI: 10.1007/s00401-017-1771-1
  • Peer Reviewed / Open Access
• [Journal Article] Spinal Cord Astrocytoma with Isocitrate Dehydrogenase 1 Gene Mutation (2017)
  • Author(s): Takai Keisuke、Tanaka Shota、Sota Takashi、Mukasa Akitake、Komori Takashi、Taniguchi Makoto
  • Journal Title: World Neurosurgery Volume: 108
  • DOI: 10.1016/j.wneu.2017.08.142
  • Peer Reviewed
• [Journal Article] The significance of histone H3 K27M mutation in brain tumors (2017)
  • Author(s): 武笠 晃丈
  • Journal Title: Prog Neuro Oncol Volume: 24 Issue: 1 Pages: 7-17
  • DOI: 10.11452/neurooncology.24.1_7
  • NAID: 130005868650
  • Peer Reviewed
• [Journal Article] Saito N. Genetic and epigenetic stability of oligodendrogliomas at recurrence. (2017)
  • Author(s): Aihara K, Mukasa A, Nagae G, Nomura M, Yamamoto S, Ueda H, Tatsuno K, Shibahara J, Takahashi M, Momose T, Tanaka S, Takayanagi S, Yanagisawa S, Nejo T, Takahashi S, Omata M, Otani R, Saito K, Narita Y, Nagane M, Nishikawa R, Ueki K, Aburatani H, Saito N
  • Journal Title: Acta Neuropathologica Communications Volume: 5 Pages: 1-11
  • DOI: 10.1186/s40478-017-0422-z
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A combination of TERT promoter mutation and MGMT methylation status predicts clinically relevant subgroups of newly diagnosed glioblastomas (2016)
  • Author(s): Arita H, Yamasaki K, Matsushita Y, Nakamura T, Shimokawa A, Takami H, Tanaka S, Mukasa A, Shirahata M, Shimizu S, Suzuki K, Saito K, Kobayashi K, Higuchi F, Uzuka T, Otani R, Tamura K, Sumita K, Ohno M, Miyakita Y, Kagawa N, Hashimoto N, Hatae R, Yoshimoto K, Shinojima N, Nakamura H, Kanemura Y, Okita Y, et al.
  • Journal Title: Acta Neuropathol Commun Volume: 4 Pages: 1-14
  • DOI: 10.1186/s40478-016-0351-2
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Diagnostic value of glutamate with 2-hydroxyglutarate in magnetic resonance spectroscopy for IDH1 mutant glioma. (2016)
  • Author(s): Nagashima H, Tanaka K, Sasayama T, Irino Y, Sato N, Takeuchi Y, Kyotani K, Mukasa A, Mizukawa K, Sakata J, Yamamoto Y, Hosoda K, Itoh T, Sasaki R, Kohmura E.
  • Journal Title: Neuro Oncol. Volume: 18(11) Pages: 1559-1568
  • DOI: 10.1093/neuonc/now090
  • Peer Reviewed
• [Presentation] Molecular characteristics of diffuse cerebellar glioma (2018)
  • Author(s): Akitake Mukasa, Masashi Nomura, Genta Nagae, Shogo Yamamoto, Kenji Tatsuno, Hiroki Ueda, Shiro Fukuda, Takayoshi Umeda, Tomonari Suzuki, Ryohei Otani, Keiichi Kobayashi, Takashi Maruyama, Shota Tanaka, Shunsaku Takayanagi, Takahide Nejo, Satoshi Takahashi, Koichi Ichimura, et al. (26人、1番目）
  • Organizer: International Conference on Brain Tumor Research & Therapy
  • Int'l Joint Research
• [Presentation] 多様なグリオーマに対する precision medicine の可能性と課題 (2018)
  • Author(s): 武笠晃丈
  • Organizer: 脳腫瘍の基礎シンポジウム
  • Invited
• [Presentation] Glioma: What does the average neurosurgeon need to know about current progress in molecular biology and how will this affect our treatment strategies? (2018)
  • Author(s): Akitake Mukasa
  • Organizer: Society of Neurosurgeons of South Africa Congress 2018
  • Int'l Joint Research / Invited
• [Presentation] マルチモダリティー時代のグリオーマ治療戦略 (2018)
  • Author(s): 武笠晃丈
  • Organizer: 日本脳腫瘍の外科学会
• [Presentation] 組織・細胞形態と分子プロファイルの多様性から考える脳腫瘍治療戦略 (2018)
  • Author(s): 武笠晃丈
  • Organizer: 日本組織細胞化学会総会・学術集会
  • Invited
• [Presentation] 脳腫瘍のゲノム医療 (2018)
  • Author(s): 武笠晃丈
  • Organizer: 日本脳神経外科学会学術総会
  • Invited
• [Presentation] グリオーマのゲノム・エピゲノム異常 (2018)
  • Author(s): 武笠晃丈
  • Organizer: 九州遺伝子診断研究会
  • Invited
• [Presentation] The characteristics of molecular profile changes in recurrent lower grade gliomas and their implication for therapeutic strategy (2017)
  • Author(s): Akitake Mukasa
  • Organizer: The 8th Institute for Refractory Cancer Research (IRCR) International Symposium
  • Place of Presentation: Samsung medical center (Seoul, Korea)
  • Year and Date: 2017-01-13
  • Int'l Joint Research / Invited
• [Presentation] The characteristics of molecular profile changes in recurrent lower grade gliomas and their implication for therapeutic strategy (2017)
  • Author(s): Akitake Mukasa, Kuniaki Saito, Shota Tanaka, Shunsaku Takayanagi,Yoshitaka Narita, Motoo Nagane, Keisuke Ueki, Ryo Nishikawa, Hiroyuki Aburatani, Nobuhito Saito
  • Organizer: 5th Quadrennial Meeting of the World Federation of Neuro-Oncology Societies
  • Int'l Joint Research
• [Presentation] Spatial Heterogeneity of Molecular Profiles in Gliomas and its Implications for Cell-of-Origin and Targeted Therapy (2017)
  • Author(s): Akitake Mukasa
  • Organizer: The 72nd Fujihara Seminar
  • Int'l Joint Research / Invited
• [Presentation] Genetic and epigenetic stability of oligodendrogliomas at recurrence (2017)
  • Author(s): Akitake Mukasa, Koki Aihara, Masashi Nomura, Shota Tanaka, Junji Shibahara, Yoshitaka Narita, Motoo Nagane, Ryo Nishikawa, Keisuke Ueki, Hiroyuki Aburatani, Nobuhito Saito
  • Organizer: The 14th Meeting of the Asian Society for Neuro-Oncology
  • Int'l Joint Research
• [Presentation] Characteristic molecular profile changes in primary and recurrent gliomas depending on their histopathology (2017)
  • Author(s): Akitake Mukasa, Masashi Nomura, Kuniaki Saito, Koki Aihara, Genta Nagae, Takahide Nejo, Shota Tanaka, Shunsaku Takayanagi, Satoshi Takahashi, Mayu Omata, Taishi Nakamura, Yoshitaka Narita, Yoshihiro Muragaki, Motoo Nagane, Keisuke Ueki, Ryo Nishikawa, Hiroyuki Aburatani, Nobuhito Saito
  • Organizer: 22nd Annual Meeting of the Society of Neuro-Oncology
  • Int'l Joint Research
• [Presentation] 再発 oligodendroglioma の ゲノム・エピゲノムの安定性 (2017)
  • Author(s): 武笠晃丈、相原功輝、野村昌志、田中將太、高柳俊作、永根基雄、成田善孝、植木敬介、油谷浩幸、斉藤延人
  • Organizer: 第34回日本脳腫瘍病理学会
• [Presentation] Integrated and comparative multi-omics analysis of primary and recurrent glioma and its clinical application (2017)
  • Author(s): Akitake Mukasa, Masashi Nomura, Kuniaki Saito, Koki Aihara, Genta Nagae, Takahide Nejo, Shunsaku Takayanagi, Shota Tanaka, Taishi Nakamura, Yoshitaka Narita, Yoshihiro Muragaki, Motoo Nagane, Keisuke Ueki, Ryo Nishikawa, Hiroyuki Aburatani, Nobuhito Saito
  • Organizer: 第76回日本脳神経外科学会
• [Presentation] 初発・再発神経膠腫のマルチオミクス比較解析から考える悪性転化機序とその治療戦略 (2017)
  • Author(s): 武笠晃丈、野村昌志、齊藤邦昭、相原功輝、永江玄太、上田 宏生、山本 尚吾、辰野 健二、根城 尭英、高橋 慧、高柳俊作、田中將太、花 大洵、北川陽介、中村大志、成田善孝、村垣善浩、永根基雄、西川 亮、植木敬介、油谷浩幸、斉藤延人
  • Organizer: 第35日本脳腫瘍学会
• [Presentation] 遺伝子プロファイル解析から考える神経膠腫の診断と治療戦略 (2016)
  • Author(s): 武笠晃丈
  • Organizer: Temozolomide Forum in HIROSHIMA 2016
  • Place of Presentation: リーガロイヤルホテル広島（広島県・広島市）
  • Year and Date: 2016-12-13
  • Invited
• [Presentation] Molecular pathological analysis on IDH-wildtype adult low-grade gliomas (2016)
  • Author(s): Akitake Mukasa, Koki Aihara, Genta Nagae, Junji Shibahara, Shota Tanaka, Shunsaku Takayanagi, Masashi Nomura, Motoo Nagane, Yoshitaka Narita, Keisuke Ueki, Ryo Nishikawa, Hiroyuki Aburatani, Nobuhito Saito
  • Organizer: 21th Annual Meeting of the Society of Neuro-Oncology
  • Place of Presentation: Farimont Scottsdate Princess Hotel (Scottsdale, Arizona, USA)
  • Year and Date: 2016-11-19
  • Int'l Joint Research
• [Presentation] Influence of anticancer therapy on clonal evolution and intratumoral heterogeneity of gliomas (2016)
  • Author(s): Akitake Mukasa
  • Organizer: 75rd Annual Meeting of the Japanese Cancer Association
  • Place of Presentation: パシフィコ横浜（神奈川県・横浜市）
  • Year and Date: 2016-10-08
  • Invited
• [Presentation] 再発神経膠腫の分子遺伝学的変化に基づく治療戦略の構築 (2016)
  • Author(s): 武笠晃丈、相原功輝、齊藤邦昭、永江玄太、野村昌志、根城 尭英、高柳俊作、田中將太、柳澤 俊介、高橋 慧、西川亮、永根基雄、成田善孝、植木敬介、油谷浩幸、斉藤延人
  • Organizer: 第75回日本脳神経外科学会
  • Place of Presentation: 福岡国際会議場（福岡県・福岡市）
  • Year and Date: 2016-09-30
• [Presentation] グリオーマのゲノム多様性とクローン進化に及ぼす抗がん治療の影響 (2016)
  • Author(s): 武笠晃丈
  • Organizer: 第21回脳腫瘍の外科学会
  • Place of Presentation: 虎ノ門ヒルズフォーラム（東京都・港区）
  • Year and Date: 2016-09-09
  • Invited
• [Book] 上衣系腫瘍, グリオーマ治療のDecision Making 脳神経外科診療プラクティス (2016)
  • Author(s): 武笠晃丈
  • Total Pages: 4
  • Publisher: 文光堂
• [Book] 脳腫瘍の部位別発症頻度と問題点, 脳腫瘍学 日本臨床 2016年9月増刊号 (2016)
  • Author(s): 武笠晃丈、田中將太、斉藤延人
  • Total Pages: 5
  • Publisher: 日本臨床社