Biological roles of ALK-binding molecules in neuroblastoma

• Project/Area Number: 26290043
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Tumor biology
• Research Institution: Kitasato University (2015-2017); National Cancer Center Japan (2014)
• Principal Investigator: Sakai Ryuichi 北里大学, 医学部, 教授 (40215603)
• Co-Investigator(Kenkyū-buntansha): 白木原 琢哉 北里大学, 医学部, 助教 (30548756)
• Research Collaborator: YAMAGUCHI Hideki 公立財団法人佐々木研究所, 腫瘍細胞研究部, 部長 ; Tomiyama Arata 防衛医科大学校, 脳神経外科, 講師
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥16,770,000 (Direct Cost: ¥12,900,000、Indirect Cost: ¥3,870,000); Fiscal Year 2016: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2015: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2014: ¥7,150,000 (Direct Cost: ¥5,500,000、Indirect Cost: ¥1,650,000)
• Keywords: 神経芽腫 / SHP2 / ALK / チロシンキナーゼ / チロシンリン酸化 / Flotillin / Shp2 / 癌 / 細胞・組織 / 分子標的治療薬 / 薬剤感受性 / ＡＬＫ / ＳＨＰ２ / Flotillin-1

Outline of Final Research Achievements

By the screening of phosphotyrosine-containing proteins associated with ALK in neuroblastoma cells, we identified SHP2 as one of the binding partners of ALK. Immunoprecipitation study revealed binding of SHP2 with ALK by ALK activity-dependent manner, and knockdown of ALK or inhibition of kinase activity of ALK by ALK inhibitors suppressed phosphorylation of SHP2 at Tyr540 and Tyr580 in NB-39-nu neuroblastoma cells which have ALK addiction. In addition, knockdown of ALK-binding docking protein ShcC resulted in decrease of ALK-SHP2 interaction. On the other hands, treatment of SHP2 inhibitor PHPS1 or knockdown of SHP2 resulted in down-regulation of ERK1/2 activation, proliferation and migration of NB-39-nu cell. However, phosphorylation of ALK was up-regulated by inhibition or knockdown of SHP2. From these results, very complicated interplay between ALK and SHP2 during the regulation of oncogenesis of the neuroblastoma cells is suggested.

Research Products

• [Journal Article] Intracellular cholesterol level regulates sensitivity of glioblastoma cells against temozolomide-induced cell death by modulation of caspase-8 activation via death receptor 5-accumulation and activation in the plasma membrane lipid raft (2018)
  • Author(s): Yamamoto Yutaro、Tomiyama Arata、Sasaki Nobuyoshi、Yamaguchi Hideki、Shirakihara Takuya、Nakashima Katsuhiko、Kumagai Kosuke、Takeuchi Satoru、Toyooka Terushige、Otani Naoki、Wada Kojiro、Narita Yoshitaka、Ichimura Koichi、Sakai Ryuichi、Namba Hiroki、Mori Kentaro
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 495 Issue: 1 Pages: 1292-1299
  • DOI: 10.1016/j.bbrc.2017.11.113
  • Peer Reviewed
• [Journal Article] Aberrant alternative splicing of RHOA is associated with loss of its expression and activity in diffuse-type gastric carcinoma cells (2018)
  • Author(s): Miyamoto Shingo、Nagamura Yuko、Nakabo Ayaka、Okabe Akira、Yanagihara Kazuyoshi、Fukami Kiyoko、Sakai Ryuichi、Yamaguchi Hideki
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 495 Issue: 2 Pages: 1942-1947
  • DOI: 10.1016/j.bbrc.2017.12.067
  • Peer Reviewed / Open Access
• [Journal Article] The p53 activator overcomes resistance to ALK inhibitors by regulating p53-target selectivity in ALK-driven neuroblastomas (2018)
  • Author(s): Makoto Miyazaki, Ryo Otomo, Yuko Matsushima-Hibiya, Hidenobu Suzuki, Ayana Nakajima, Arata Tomiyama, Koichi Ichimura, Koichi Matsuda, Toshiki Watanabe, Takahiro Ochiya, Hitoshi Nakagama, Ryuichi Sakai, Yoichi Taya, and Masato Enari
  • Journal Title: Cell Death Discovery Volume: -
  • Peer Reviewed
• [Journal Article] Novel small molecule inhibiting CDCP1-PKCδ pathway reduces tumor metastasis and proliferation. (2017)
  • Author(s): Nakashima K, Uekita T, Yano S, Kikuchi JI, Nakanishi R, Sakamoto N, Fukumoto K, Nomoto A, Kawamoto K, Shibahara T, Yamaguchi H, Sakai R.
  • Journal Title: Cancer Sci. Volume: 印刷中 Issue: 5 Pages: 1049-1057
  • DOI: 10.1111/cas.13218
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Actinin-1 and actinin-4 play essential but distinct roles in invadopodia formation by carcinoma cells (2017)
  • Author(s): Yamaguchi Hideki、Ito Yuumi、Miura Nami、Nagamura Yuko、Nakabo Ayaka、Fukami Kiyoko、Honda Kazufumi、Sakai Ryuichi
  • Journal Title: European Journal of Cell Biology Volume: 96 Issue: 7 Pages: 685-694
  • DOI: 10.1016/j.ejcb.2017.07.005
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Report of the Japan diabetes society/Japanese cancer association joint committee on diabetes and cancer, Second report. (2016)
  • Author(s): Goto A, Noto H, Noda M, Ueki K, Kasuga M, Tajima N, Ohashi K, Sakai R, Tsugane S, Hamajima N, Tajima K, Imai K, Nakagama H.
  • Journal Title: Cancer Science Volume: 107(3) Issue: 3 Pages: 369-71
  • DOI: 10.1111/cas.12889
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Recurrent neomorphic mutations of MTOR in central nervous system and testicular germ cell tumors may be targeted for therapy (2016)
  • Author(s): Ichimura K, Fukushima S, Nagane M, The Intracranial Berm Cell Tumor Genome Analysis Consortium, et al
  • Journal Title: Acta Neuropathol Volume: Published online: 08 March Issue: 6 Pages: 889-901
  • DOI: 10.1007/s00401-016-1557-x
  • Peer Reviewed
• [Journal Article] Direct interaction between carcinoma cells and cancer associated fibroblasts for the regulation of cancer invasion (2015)
  • Author(s): Yamaguchi H, and Sakai R
  • Journal Title: Cancers Volume: 7 Issue: 4 Pages: 2054-2062
  • DOI: 10.3390/cancers7040876
  • Peer Reviewed / Open Access
• [Journal Article] Augmentation of invadopodia formation in temozolomide-resistant or adopted glioma is regulated by c-Jun terminal kinase-paxillin axis (2015)
  • Author(s): Ueno H, Tomiyama A, Yamaguchi H, Uekita T et al.
  • Journal Title: Biochem Biophys Res Commun Volume: 468 Issue: 1-2 Pages: 240-247
  • DOI: 10.1016/j.bbrc.2015.10.122
  • Peer Reviewed
• [Journal Article] Oncogenic Ras/ERK signaling activates CDCP1 to promote tumor invasion and metastasis (2014)
  • Author(s): Uekita T, Fujii S et al.
  • Journal Title: Molecular Cancer Research Volume: 12 Issue: 10 Pages: 1449-1459
  • DOI: 10.1158/1541-7786.mcr-13-0587
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Stromal fibroblasts mediate extracellular matrix remodeling and invasion of scirrhous gastric carcinoma cells. (2014)
  • Author(s): Yamaguchi H, Yoshida H, Takanashi M, Ito Y, Fukami K, Yanagihara K, Yashiro M, Sakai R.
  • Journal Title: PlosOne Volume: 9 Issue: 1 Pages: e85485-e85485
  • DOI: 10.1371/journal.pone.0085485
  • Peer Reviewed / Open Access
• [Journal Article] Saracatinib impairs the peritoneal dissemination of diffuse-type gastric carcinoma cells resistant to Met and FGFR inhibitors. (2014)
  • Author(s): Yamaguchi H, Takanashi M, Yoshida N, Ito Y, Kamata R, Fukami K, Yanagihara K, Sakai R.
  • Journal Title: Cancer Sci. Volume: 105 Issue: 5 Pages: 528-536
  • DOI: 10.1111/cas.12387
  • Peer Reviewed / Open Access
• [Journal Article] Flotillin-1 regulates oncogenic signaling in neuroblastoma cells by regulating ALK membrane association. (2014)
  • Author(s): Tomiyama A, Uekita T, Kamata R, Sasaki K, Takita J, Ohira M, Nakagawara A, Kitanaka C, Mori K, Yamaguchi H, Sakai R.
  • Journal Title: Cancer Res. Volume: 74(14) Issue: 14 Pages: 3790-3801
  • DOI: 10.1158/0008-5472.can-14-0241
  • Peer Reviewed / Open Access
• [Presentation] スキルス胃癌細胞におけるSHP2の機能解析 (2017)
  • Author(s): 山口英樹、富山新太、堺隆一
  • Organizer: 第７６回日本癌学会学術総会
• [Presentation] The signaling complex of tyrosine phosphstase SHP2 and docking protein ShcC regulates oncogenicity of neuroblastoma cells in a tyrosine-phosphorylation dependent manner (2017)
  • Author(s): Arata Tomiyama and Ryuichi Sakai
  • Organizer: The 3rd Asia-Pacific International Symposium of Neuroblastoma
  • Int'l Joint Research
• [Presentation] Interaction of SHP2 with ALK regulates oncogenicity of neuroblastoma cells (2016)
  • Author(s): Ryuichi Sakai
  • Organizer: 12th International Conference on Protein Phosphatase
  • Place of Presentation: 近畿大学（大阪府東大阪市）
  • Year and Date: 2016-10-27
  • Int'l Joint Research
• [Presentation] 神経芽腫におけるShcCを介したSHP2とALKの結合は、神経芽腫細胞の腫瘍原性を制御する (2016)
  • Author(s): Arata Tomiyama, Takuya Shirakihara, Katsuhiko Nakashima, Hideki Yamaguchi, Kentaro Mori, Ryuichi Sakai
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-10-06
• [Presentation] The signaling complex of tyrosine phosphatase SHP2 and docking protein ShcC regulates oncogenicity of neuroblastoma cells in a tyrosine-phosphorylation dependent manner. (2016)
  • Author(s): Arata Tomiyama, Ryuichi Sakai
  • Organizer: Advances in Neuroblastoma Research 2016
  • Place of Presentation: Cairns, Australia
  • Year and Date: 2016-06-19
  • Int'l Joint Research
• [Presentation] 神経芽腫におけるALK受容体シグナルのFlotillin-1とShcC-Shp2経路による制御 (2016)
  • Author(s): 堺隆一、富山新太
  • Organizer: 第7回日本プロテインホスファターゼ研究会学術集会
  • Place of Presentation: 自然科学研究機構山手地区（愛知県岡崎市）
  • Year and Date: 2016-01-29
• [Presentation] 神経芽腫のALK阻害剤感受性を制御するALK結合リン酸化蛋白質群 (2015)
  • Author(s): 堺隆一、富山新太
  • Organizer: 第38回日本分子生物学会年会第88回日本生化学大会・合同大会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
• [Presentation] Identification of ALK-binding phosphotyrosine-containing proteins as the critical regulators of oncogenic properties of neuroblastoma cells (2015)
  • Author(s): 富山新太、堺隆一
  • Organizer: 2015 Asia-Pacific Symposium of Neuroblasoma
  • Place of Presentation: 台北（台湾）
  • Year and Date: 2015-11-14
  • Int'l Joint Research / Invited
• [Presentation] 神経芽腫細胞におけるSHP2とALKの結合は、神経芽腫の腫瘍原性を制御する (2015)
  • Author(s): 富山新太、上野英明、白木原琢哉、中島克彦、山口英樹、森健太郎、堺隆一
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場（愛知県名古屋市）
  • Year and Date: 2015-10-10
• [Presentation] Regulation of oncogenic signaling in neuroblastoma through binding partners of anaplastic lymphoma kinase. (2014)
  • Author(s): Sakai R
  • Organizer: 第11回プロテインホスファターゼ国際カンファレンス
  • Place of Presentation: 仙台
  • Year and Date: 2014-11-12 – 2014-11-14
• [Presentation] 新規ALK結合蛋白質であるFlotillin-1は、ALKの細胞膜結合の調節を介してALKシグナルを制御する． (2014)
  • Author(s): 富山新太，上北尚正，山口英樹，上野英明，滝田順子，佐々木一樹，中川原章，森健太郎，堺隆一
  • Organizer: 第73回日本癌学会学術総会
  • Place of Presentation: 横浜
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Flotillin-1 regulates oncogenic signaling in neuroblastoma cells through receptor endocytosis of anaplastic lymphoma kinase (2014)
  • Author(s): Tomiyama A，Uekita T，Sakai R
  • Organizer: Advances in Neuroblastoma Research 2014
  • Place of Presentation: ケルン、ドイツ
  • Year and Date: 2014-05-13 – 2014-05-16
• [Remarks] 新領域創成グループ 難治進行がん研究分野
  • URL: http://www.ncc.go.jp/jp/nccri/divisions/rac/index.html