Analysis of heterogeneity of tumor stem cells and EMT using Crk adaptor protein

• Project/Area Number: 15K15106
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Experimental pathology
• Research Institution: Hokkaido University
• Principal Investigator: Tsuda Masumi 北海道大学, 医学研究科, 講師 (30431307)
• Co-Investigator(Kenkyū-buntansha): 谷野 美智枝 北海道大学, 医学研究科, 講師 (90360908)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: エクソソーム / CRK / 浸潤 / 転移 / 腫瘍ヘテロジェナイティ / 癌 / ヘテロジェナイティー / Crk / EMT

Outline of Final Research Achievements

We unveiled a novel function of CRK adaptors via exosomes in tumor progression and metastasis in invasive bladder cancer (BC). CRK is overexpressed in invasive UM-UC-3 BC cells, and CRK knockdown downregulated the expression of ErbB2. In an orthotopic xenograft model, metastases to lung, liver, and bone of UM-UC- 3 cells were completely abolished by CRK elimination. Mass spectrometry analysis identified that ErbB2 is contained in UM-UC-3-derived exosomes, which significantly increased proliferation and invasion of low-grade 5637 BC cells and HUVECs. In athymic mice educated by UM-UC-3-derived exosomes, intravenously implanted UM-UC-3 cells developed lung metastasis with increased tumor angiogenesis, whereas exosomes with CRK depletion could not. In conclusion, CRK adaptors contribute to tumor heterogeneity of invasive BC via exosome, resulting in the tumor progression and metastasis.

Research Products

• [Journal Article] Notch1 regulates invasion and metastasis of head and neck squamous cell carcinoma by inducing EMT through c-Myc. (2017)
  • Author(s): Inamura N, Kimura T, Wang L, Yanagi H, Tsuda M, Tanino M, Nishihara H, Fukuda S, Tanaka S.
  • Journal Title: Auris Nasus Larynx Volume: 44 Issue: 4 Pages: 447-457
  • DOI: 10.1016/j.anl.2016.08.003
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Clinicopathological evaluation of Sox10 expression in diffuse-type gastric adenocarcinoma. (2017)
  • Author(s): Kato M, Nishihara H, Hayashi H, Kimura T, Ishida Y, Wang L, Tsuda M, Tanino MA, Tanaka S.
  • Journal Title: Med Oncol. Volume: 34 Pages: 8-8
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Preclinical evidence of anti-tumor activity induced by EZH2 inhibition in human models of synovial sarcoma. (2016)
  • Author(s): Kawano S, Grassian AR, Tsuda M, Knutson SK, Warholic NM, Kuznetsov G, Xu S, Xiao Y, Pollock RM, Smith JS, Kuntz KK, Ribich S, Minoshima Y, Matsui J, Copeland RA, Tanaka S, Keihack H.
  • Journal Title: PLoS One Volume: 11 Issue: 1 Pages: e0170539-e0170539
  • DOI: 10.1371/journal.pone.0170539
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Aldo-keto reductase 1C1 induced by interleukin-1beta mediates the invasive potential and drug resistance of metastatic bladder cancer cells. (2016)
  • Author(s): Matsumoto R, Tsuda M, Yoshida K, Tanino M, Kimura T, Nishihara H, Abe T, Shinohara N, Nonomura K, Tanaka S
  • Journal Title: Scientific reports Volume: 4 Issue: 1 Pages: 34625-34625
  • DOI: 10.1038/srep34625
  • Peer Reviewed / Open Access
• [Journal Article] Rapid immunocytochemistry based on alternating current electric field using squash smear preparation of central nervous system tumors. (2016)
  • Author(s): Moriya J, Tanino AM, Takenami T, Endoh T, Urushido M, Kato Y, Wang L, Kimura T, Tsuda M, Nishihara H, Tanaka S, R-IHC study Group
  • Journal Title: Brain Tumor Pathol Volume: 33 Issue: 1 Pages: 13-18
  • DOI: 10.1007/s10014-015-0238-0
  • NAID: 120005905603
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification and analysis of CXCR4-positive synovial sarcoma initiating cells. (2016)
  • Author(s): Kimura T, Wang L, Tabu K, Tsuda M, Tanino M, Maekawa A, Nishihara H, Hiraga H, Taga T, Oda Y, Tanaka S.
  • Journal Title: Oncogene Volume: 未定 Issue: 30 Pages: 3932-3943
  • DOI: 10.1038/onc.2015.461
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Synthetic PAMPS gel activates BMP/Smad signaling pathway in ATDC5 cells, which plays a significant role in the gel-induced chondrogenic differentiation. (2016)
  • Author(s): Goto K, Kimura T, Kitamura N, Semba S, Ohmiya Y, Aburatani S, Matsukura S, Tsuda M, Kurokawa T, Gong JP, Tanaka S, Yasuda K.
  • Journal Title: J Biomed Mater Res A Volume: 未定 Issue: 3 Pages: 734-746
  • DOI: 10.1002/jbm.a.35615
  • NAID: 120005981343
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Attenuation of ligand-induced activation of angiotensin II type 1 receptor signaling by the type 2 receptor via protein kinase C (2016)
  • Author(s): Inuzuka T, Fujioka Y, Tsuda M, Fujioka M, Satoh AO, Horiuchi K, Nishide S, Nanbo A, Tanaka S, Ohba Y
  • Journal Title: Sci Rep Volume: 6 Issue: 1 Pages: 21613-21613
  • DOI: 10.1038/srep21613
  • NAID: 120005722549
  • Peer Reviewed / Open Access
• [Journal Article] Analysis of NAB2-STAT6 gene fusion in 17 cases of meningeal solitary fibrous tumor/hemangiopericytoma: review of the literature. (2016)
  • Author(s): Yuzawa S, Nishihara H, Wang L, Tsuda M, Kimura T, Tanino M, Tanaka S.
  • Journal Title: Am J Surg Pathol Volume: 未定
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Receptor activator of NF-κB ligand induces cell adhesion and integrin α2 expression via NF-κB in head and neck cancers (2016)
  • Author(s): T.Yamada, M. Tsuda, T. Wagatsuma, Y. Fujioka, M. Fujioka, A. O. Satoh, K. Horiuchi, S. Nishide, A. Nanbo, Y. Totsuka, H. Haga, S. Tanaka, M. Shindoh, and Y. Ohba
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 23545-23545
  • DOI: 10.1038/srep23545
  • NAID: 120005753715
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Novel signaling collaboration between TGF-beta and adaptor protein Crk facilitates EMT in human lung cancer (2016)
  • Author(s): Elmansuri AZ, Tanino MA, Mahabir R, Wang L, Kimura T, Nishihara H, Kinoshita I, Dosaka-Akita H, Tsuda M and Tanaka S
  • Journal Title: Oncotarget Volume: （in press） Issue: 19 Pages: 27094-107
  • DOI: 10.18632/oncotarget.8314
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Clinical impact of targeted amplicon sequence for meningioma as a practical clinical sequence system. (2016)
  • Author(s): Yuzawa S, Nishihara H, Yamaguchi S, Mohri H, Wang L, Kimura T, Tsuda M, Tanino M, Kobayashi H, Terasaka S, Houkin K, Sato N, Tanaka S.
  • Journal Title: Mod Pathol Volume: 未定
  • NAID: 120005905622
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Clinicopathological Study of Pilomyxoid-Spectrum Astrocytomas:An Analysis of the BRAF Gene. Report of Two Cases (2015)
  • Author(s): Ito T, Sato K, Oikawa M, Sugio H, Asanome T, Ozaki Y, Nakamura H, Tanaka S, Tsuda M, Nagashima K
  • Journal Title: Neurological Surgery 脳神経外科 Volume: 43 Issue: 9 Pages: 825-833
  • DOI: 10.11477/mf.1436203129
  • ISSN: 0301-2603, 1882-1251
  • Year and Date: 2015-09-10
  • Peer Reviewed
• [Journal Article] CRK adaptor protein induces epithelial-mesenchymal transition and metastasis of bladder cancer cells via HGF/c-Met feedback loop. (2015)
  • Author(s): Matsumoto R, Tsuda M, Wang L, Maishi N, Abe t, Kimura T, Tanino M, Nishihara H, Hida K, Ohba Y, Shinohara N, Nonomura K, Tanaka S.
  • Journal Title: Cancer Sci Volume: in press Issue: 6 Pages: 709-17
  • DOI: 10.1111/cas.12662
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive glycomics of a multistep human brain tumor model reveals specific glycosylation patterns related to malignancy. (2015)
  • Author(s): Furukawa J, Tsuda M, Okada K, Kimura T, Piao J, Tanaka S, Shinohara Y.
  • Journal Title: PLoS One Volume: 10 Issue: 7 Pages: e0128300-e0128300
  • DOI: 10.1371/journal.pone.0128300
  • NAID: 120005650133
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Tyr724 phosphorylation of ELMO1 by Src is involved in cell spreading and migration via Rac1 activation. (2015)
  • Author(s): Y. Makino, M. Tsuda, Y. Ohba, H. Nishihara, H. Sawa, K. Nagashima and S. Tanaka.
  • Journal Title: Cell Commun. Signal. Volume: 13 Issue: 1 Pages: 42-42
  • DOI: 10.1186/s12964-015-0113-y
  • NAID: 120005651855
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] AKR1C1 mediates bladder cancer metastasis and drug resistance. (2016)
  • Author(s): 津田真寿美、松本隆児、吉田一彦、谷野美智枝、木村太一、西原広史、阿部崇重、篠原信雄、野々村克也、田中伸哉
  • 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] NGS-based MSK-IMPACT analysis reveals specific genetic alterations in recurrent glioblastoma. (2015)
  • Author(s): Tsuda M, Kohsaka S, Wang L, Kimura T, Tanino M, Nishihara H, Ladanyi M, Tanaka S
  • Organizer: 20th Annual Society for Neuro-Oncology Annual Scientific Meeting
  • Place of Presentation: San Antonio, USA
  • Year and Date: 2015-11-19
  • Int'l Joint Research
• [Presentation] NGS-based MSK-IMPACT analysis reveals specific genetic alterations in recurrent glioblastoma. (2015)
  • Author(s): 津田真寿美、高阪真路、王磊、木村太一、谷野美智枝、西原広史、Marc Ladanyi、田中伸哉
  • Organizer: 第74回日本癌学会総会
  • Place of Presentation: 名古屋国際会議場（愛知県・名古屋市）
  • Year and Date: 2015-10-08
• [Presentation] 膠芽腫における分子標的阻害薬耐性獲得責任分子としてのIGFBP2の同定 (2015)
  • Author(s): 津田真寿美、王磊、谷野美智枝、木村太一、西原広史、田中伸哉
  • Organizer: 第33回日本脳腫瘍病理学会
  • Place of Presentation: JRホテルクレメント高松（香川県・高松市）
  • Year and Date: 2015-05-29
• [Remarks] http://patho2.med.hokudai.ac.jp/
• [Patent(Industrial Property Rights)] 癌幹細胞の製造方法 (2017)
  • Inventor(s): 田中伸哉、安田和則、グン剣萍、津田真寿美、黒川孝幸
  • Industrial Property Rights Holder: 北海道大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2017-02-20
• [Patent(Industrial Property Rights)] 固定生体組織内での活性型低分子量GTP結合蛋白質検出方法 (2016)
  • Inventor(s): 田中伸哉、津田真寿美、谷野美智枝
  • Industrial Property Rights Holder: 北海道大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-03-15
  • Overseas
• [Patent(Industrial Property Rights)] グリオーマの診断マーカー、診断方法、糖鎖マーカーを検出する方法及び糖鎖マーカー (2015)
  • Inventor(s): 篠原康郎、田中伸哉、古川潤一、津田真寿美
  • Industrial Property Rights Holder: 北海道大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-08-07
  • Overseas