In vivo characterization of carcinogenesis and cancer-host interaction in colorectal cancer using two photon laser scanning microscopy

• Project/Area Number: 21390377
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: Mie University
• Principal Investigator: KUSUNOKI Masato 三重大学, 医学(系)研究科(研究院), 教授 (50192026)
• Co-Investigator(Kenkyū-buntansha): INOUE Yasuhiro 三重大学, 医学部附属病院, 講師 (20324535) ; MIKI Chikao 三重大学, 大学院・医学系研究科, 客員教授 (50242962) ; TOIYAMA Yuji 三重大学, 大学院・医学系研究科, 助教 (00422824)
• Project Period (FY): 2009-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥12,480,000 (Direct Cost: ¥9,600,000、Indirect Cost: ¥2,880,000); Fiscal Year 2013: ¥520,000 (Direct Cost: ¥400,000、Indirect Cost: ¥120,000); Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2009: ¥8,060,000 (Direct Cost: ¥6,200,000、Indirect Cost: ¥1,860,000)
• Keywords: 二光子レーザー顕微鏡 / 癌幹細胞 / 癌-間質反応 / 二光子励起顕微鏡 / 大腸癌 / 肝転移 / 上皮間葉移行 / 大陽癌

Research Abstract

We attempted to visualize in vivo dynamic interactions between cancer stem cells (CSCs) and their surrounding stromal cells during the process of carcinogenesis, tumor invasion and metastasis in colorectal cancer using two photon laser scanning microscopy (TPLSM). We hypothesized that residual cancer cells after chemoradiotherapy (CRT) may contain putative CSCs which show not only CRT-resistant property, but also epithelial-mesenchymal transition (EMT) phenotype.
On liver metastasis model, dynamic tumor-host interactions including tumor-endothelial, tumor-platelet, and tumor-leukocyte interactions were observed in vivo real-time by TPLSM. Treatment resistant clones (made by repeated exposure of anti-cancer drug or irradiation) were inoculated into mice, and visualized their characteristics in vivo real-time using TPLSM. The small fraction of treatment resistant clones showed spindle shaped morphology. Their growth pattern was changed from localized growth to diffusely invasive growth.

Research Products

• [Journal Article] Serum miR-200c is a novel prognostic and metastasis-predictive biomarker in patients with colorectal cancer (2014)
  • Author(s): Toiyama Y, Hur K, Tanaka K, Inoue Y, Kusunoki M, Boland CR, Goel A
  • Journal Title: Ann Surg Volume: 259(4) Pages: 735-743
  • Peer Reviewed
• [Journal Article] C-reactive protein as predictor of recurrence in patients with rectal cancer undergoing chemoradiotherapy followed by surgery (2013)
  • Author(s): Toiyama Y, Inoue Y, Saigusa S, Kawamura M, Kawamoto A, Okugawa Y, Hiro J, Tanaka K, Mohri Y, Kusunoki M
  • Journal Title: Anticancer Res Volume: 33(11) Pages: 5065-5074
  • Peer Reviewed
• [Journal Article] Multimodality therapy in fistula-associated perianal mucinous adenocarcinoma (2013)
  • Author(s): Inoue Y, Kawamoto A, Okigami M, Okugawa Y, Hiro J, Toiyama Y, Tanaka K, Uchida K, Mohri Y, Kusunoki M
  • Journal Title: Am Surg Volume: 79(9)
  • Peer Reviewed
• [Journal Article] Increased expression of Slug and Vimentin as novel predictive biomarkers for lymph node metastasis and poor prognosis in colorectal cancer (2013)
  • Author(s): Toiyama Y, Yasuda H, Saigusa S, Tanaka K, Inoue Y, Goel A, Kusunoki M
  • Journal Title: Carcinogenesis Volume: 34(11) Pages: 2548-2557
  • Peer Reviewed
• [Journal Article] Advances and directions in chemotherapy using implantable port systems for colorectal cancer : a historical review (2013)
  • Author(s): Inoue Y, Kusunoki M
  • Journal Title: Surg Today
  • Peer Reviewed
• [Journal Article] Leucine-rich repeat-containing G protein-coupled receptor 5 expression in ductular reactions after chemotherapy for metastatic colorectal cancer (2013)
  • Author(s): Saigusa S, Tanaka K, Toiyama Y, Matsushita K, Kawamura M, Okugawa Y, Uchida K, Inoue Y, Mohri Y, Kusunoki M
  • Journal Title: Hepatol Res Volume: 43(1) Pages: 84-90
  • NAID: 10031183461
  • Peer Reviewed
• [Journal Article] Intravital imaging of gastrointestinal diseases in preclinical models using two-photon laser scanning microscopy (2013)
  • Author(s): Tanaka K, Toiyama Y, Inoue Y, Uchida K, Araki T, Mohri Y, Mizoguchi A, Kusunoki M
  • Journal Title: Surg Today Volume: 43(2) Pages: 123-129
  • NAID: 10031167976
  • Peer Reviewed
• [Journal Article] Advances and directions in chemotherapy using implantable port systems for colorectal cancer: a historical review. (2013)
  • Author(s): Inoue Y, Kusunoki M.
  • Journal Title: Surg Today. Volume: Jul 28
  • Peer Reviewed
• [Journal Article] Increased expression of Slug and Vimentin as novel predictive biomarkers for lymph node metastasis and poor prognosis in colorectal cancer. (2013)
  • Author(s): Toiyama Y, Yasuda H, Saigusa S, Tanaka K, Inoue Y, Goel A, Kusunoki M.
  • Journal Title: Carcinogenesis. Volume: 34 Issue: 11 Pages: 2548-2557
  • DOI: 10.1093/carcin/bgt282
  • Peer Reviewed
• [Journal Article] Serum miR-21 as a diagnostic and prognostic biomarker in colorectal cancer. (2013)
  • Author(s): Toiyama Y, Takahashi M, Hur K, Nagasaka T, Tanaka K, Inoue Y, Kusunoki M, Boland CR, Goel A.
  • Journal Title: J Natl Cancer Inst. Volume: 105 Issue: 12 Pages: 849-859
  • DOI: 10.1093/jnci/djt101
  • Peer Reviewed
• [Journal Article] Intravital imaging of gastrointestinal diseases in preclinical models using two-photon laser scanning microscopy. (2013)
  • Author(s): Tanaka K, Toiyama Y, Inoue Y, Uchida K, Araki T, Mohri Y, Mizoguchi A, Kusunoki M.
  • Journal Title: Surg Today. Volume: 43 Issue: 2 Pages: 123-129
  • DOI: 10.1007/s00595-012-0283-9
  • NAID: 10031167976
  • Peer Reviewed
• [Journal Article] The preventive effects of low-dose enteric-coated aspirin tablets on the development of colorectal tumours in Asian patients: a randomised trial. (2013)
  • Author(s): Ishikawa H, Mutoh M, Suzuki S, Tokudome S, Saida Y, Abe T, Okamura S, Tajika M, Joh T, Tanaka S, Kudo SE, Matsuda T, Iimuro M, Yukawa T, Takayama T, Sato Y, Lee K, Kitamura S, Mizuno M, Sano Y, Gondo N, Sugimoto K, Kusunoki M, Goto C, Matsuura N, Sakai T, Wakabayashi K.
  • Journal Title: Gut. Volume: Jan 31 Issue: 11 Pages: 1755-1759
  • DOI: 10.1136/gutjnl-2013-305827
  • Peer Reviewed
• [Journal Article] Allelic Imbalance at an 8q24 Oncogenic SNP is Involved in Activating MYC in Human Colorectal Cancer. (2013)
  • Author(s): Sugimachi K, Niida A, Yamamoto K, Shimamura T, Imoto S, Iinuma H, Shinden Y, Eguchi H, Sudo T, Watanabe M, Tanaka J, Kudo S, Hase K, Kusunoki M, Yamada K, Shimada Y, Sugihara K, Maehara Y, Miyano S, Mori M, Mimori K.
  • Journal Title: Ann Surg Oncol. Volume: Jan 6.
• [Journal Article] Clinical significance of LGR5 and CD44 expression in locally advanced rectal cancer after preoperative chemoradiotherapy (2012)
  • Author(s): Saigusa S, Inoue Y, Tanaka K, Toiyama Y, Matsushita K, Kawamura M, Okugawa Y, Hiro J, Uchida K, Mohri Y, Kusunoki M
  • Journal Title: Int J Oncol Volume: 41(5) Pages: 1643-4652
  • Peer Reviewed
• [Journal Article] In vivo real-time imaging of chemotherapy response on the liver metastatic tumor microenvironment using multiphoton microscopy (2012)
  • Author(s): Tanaka K, Okigami M, Toiyama Y, Morimoto Y, Matsushita K, Kawamura M, Hashimoto K, Saigusa S, Okugawa Y, Inoue Y, Uchida K, Araki T, Mohri Y, Mizoguchi A, Kusunoki M
  • Journal Title: Oncol Rep Volume: 28(5) Pages: 1822-1830
  • Peer Reviewed
• [Journal Article] High TrkB expression levels are associated with poor prognosis and EMT induction in colorectal cancer cells (2012)
  • Author(s): Fujikawa H, Tanaka K, Toiyama Y, Saigusa S, Inoue Y, Uchida K, Kusunoki M
  • Journal Title: J Gastroenterol Volume: 47(7) Pages: 775-784
  • NAID: 10030962144
  • Peer Reviewed
• [Journal Article] In vivo time-course imaging of tumor angiogenesis in colorectal liver metastases in the same living mice using two-photon laser scanning microscopy (2012)
  • Author(s): Tanaka K, Morimoto Y, Toiyama Y, Matsushita K, Kawamura M, Koike Y, Okugawa Y, Inoue Y, Uchida K, Araki T, Mizoguchi A, Kusunoki M
  • Journal Title: J Oncol Volume: 2012 Pages: 265487-265487
  • NAID: 120003596966
  • Peer Reviewed
• [Journal Article] Radiation induces epithelial-mesenchymal transition in colorectal cancer cells (2012)
  • Author(s): Kawamoto A, Yokoe T, Tanaka K, Saigusa S, Toiyama Y, Yasuda H, Inoue Y, Miki C, Kusunoki M
  • Journal Title: Oncol Rep Volume: 27(1) Pages: 51-57
  • Peer Reviewed
• [Journal Article] Intravital dual-colored visualization of colorectal liver metastasis in living mice using two photon laser scanning microscopy (2012)
  • Author(s): Tanaka K, Morimoto Y, Toiyama Y, Okugawa Y, Inoue Y, Uchida K, Kimura K, Mizoguchi A, Kusunoki M
  • Journal Title: Microsc Res Tech Volume: 75(3) Pages: 307-315
  • Peer Reviewed
• [Journal Article] In vivo real-time imaging of chemotherapy response on the liver metastatic tumor microenvironment using multiphoton microscopy. (2012)
  • Author(s): Tanaka K et al.
  • Journal Title: Oncol Rep. Volume: 28 Issue: 5 Pages: 1822-1830
  • DOI: 10.3892/or.2012.1983
  • Peer Reviewed
• [Journal Article] In vivo time-course imaging of tumor angiogenesis in colorectal liver me tastases in the same living mice using two-photon laser scanning microscopy (2012)
  • Author(s): Tanaka K, Morimoto Y, Toiyama Y, Matsushita K, Kawamura M, Koike Y, Okugawa Y, Inoue Y, Uchida K, Araki T, Mizoguchi A, Kusunoki M
  • Journal Title: J Oncol Volume: 2012 Pages: 265487-265487
  • NAID: 120003596966
  • Peer Reviewed
• [Journal Article] Cancer-associated fibroblasts correlate with poor prognosis in rectal cancer after chemoradiotherapy (2011)
  • Author(s): Saigusa S, Toiyama Y, Tanaka K, Yokoe T, Okugawa Y, Fujikawa H, Matsusita K, Kawamura M, Inoue Y, Miki C, Kusunoki M
  • Journal Title: Int J Oncol Volume: 38(3) Pages: 655-663
  • Peer Reviewed
• [Journal Article] CXCL5, a promoter of cell proliferation, migration and invasion, is a novel serum prognostic marker in patients with colorectal cancer (2011)
  • Author(s): Kawamura M, Toiyama Y, Tanaka K, Saigusa S, Okugawa Y, Hiro J, Uchida K, Mohri Y, Inoue Y, Kusunoki M
  • Journal Title: Eur J Cancer
  • Peer Reviewed
• [Journal Article] Intravital imaging of DSS-induced cecal mucosal damage in GFP-transgenic mice using two-photon microscopy. (2010)
  • Author(s): Toiyama Y, Mizoguchi A, Okugawa Y, Koike Y, Morimoto Y, Araki T, Uchida K, Tanaka K, Nakashima H, Hibi M, Kimura K, Inoue Y, Miki C, Kusunoki M.
  • Journal Title: J Gastroenterol. Volume: 45 Pages: 544-553
  • NAID: 120002777869
  • Peer Reviewed
• [Journal Article] Resection of rectal cancer : a historical review. (2010)
  • Author(s): Inoue Y, Kusunoki M.
  • Journal Title: Surg Today. Volume: 40 Pages: 501-506
  • NAID: 10027395844
  • Peer Reviewed
• [Journal Article] Loss of tumoral expression of soluble IL-6 receptor is associated with disease progression in colorectal cancer. (2010)
  • Author(s): Okugawa Y, Miki C, Toiyama Y, Yasuda H, Yokoe T, Saigusa S, Hiro J, Tanaka K, Inoue Y, Kusunoki M.
  • Journal Title: Br J Cancer. Volume: 103 Pages: 787-795
  • Peer Reviewed
• [Journal Article] Microdissection is essential for gene expression analysis of irradiated rectal canter tissues (2009)
  • Author(s): Inoue Y, Tanaka K, Yokoe T, Saigusa S, Toiyama Y, Miki C, Kusunoki M.
  • Journal Title: Oncol Rep. 22 Pages: 901-906
  • Peer Reviewed
• [Journal Article] Elevated CD133, but not VEGF or EGFR, as a predictive marker of distant recurrence after preoperative chemoradiotherapy in rectal cancer (2009)
  • Author(s): Yasuda H, Tanaka K, Saigusa S, Toiyama Y, Koike Y, Okugawa Y, Yokoe T, Kawamoto A, Inoue Y, Miki C, Kusunoki M.
  • Journal Title: Oncol Rep. 22 Pages: 709-717
  • Peer Reviewed
• [Journal Article] Correlation of CD133, OCT4, and SOX2 in rectal cancer and their association with distant recurrence after chemoradiotherapy (2009)
  • Author(s): Saigusa S, Tanaka K, Toiyama Y, Yokoe T, Okugawa Y, Ioue Y, Miki C, Kusunoki M.
  • Journal Title: Ann Surg Oncol. 16 Pages: 3488-3498
  • Peer Reviewed
• [Presentation] Clinical significance of serum angiopoietin-like protein 2 in colorectal cancer patients (2014)
  • Author(s): Takahito Kitajima, Yuji Toiyama, Tadanobu Shimura, Shozo Ide, Hiroki Imaoka, Satoru Kondo, Mikio Kawamura, Yoshinaga Okugawa, Aya Kawamoto, Junichiro Hiro, Susumu Saigusa, Masaki Ohi, Koji Tanaka, Yasuhiro Inoue, Yasuhiko Mohri, Masato Kusunoki
  • Organizer: 2014 Gastrointestinal Cancers Symposium
  • Place of Presentation: San Francisco, CA, USA
• [Presentation] Serum angiopoietin-like protein 2 as a novel biomarker of diagnosis and prognosis in gastric cancer (2014)
  • Author(s): Yuji Toiyama, Yasuhiro Inoue, Takahito Kitajima, Tadanobu Shimura, Hiroki Imaoka, Satoru Kondo, Shozo Ide, Masato Okigami, Hiromi Yasuda, Susumu Saigusa, Masaki Ohi, Koji Tanaka, Yasuhiko Mohri, Masato Kusunoki
  • Organizer: 2014 Gastrointestinal Cancers Symposium
  • Place of Presentation: San Francisco, CA, USA
• [Presentation] The clinical significance of tissue miR-124 methylation in sporadic and ulcerative colitis-associated colorectal cancer (2013)
  • Author(s): Yuji Toiyama, Keun Hur, Koji Tanaka, Yasuhiro Inoue, Takeshi Nagasaka, Masato Kusunoki, C. Richard Boland, Ajay Goel
  • Organizer: AACR Annual Meeting 2013
  • Place of Presentation: Chicago, Illinois, USA
• [Presentation] Prognostic value of angiopoietin-like protein 2 (ANGPTL2) at the tumor margins in patients with gastric cancer (2013)
  • Author(s): Tadanobu Shimura, Koji Tanaka, Susumu Saigusa, Satoru Kondo, Takahito Kitajima, Yuji Toiyama, Yoshinaga Okugawa, Yasuhiro Inoue, Toshimitsu Araki, Keiichi Uchida, Yasuhiko Mohri, Tsutomu Nobori, Masato Kusunoki
  • Organizer: AACR Annual Meeting 2013
  • Place of Presentation: Chicago, Illinois, USA
• [Presentation] Expression of tropomyosin-related kinase B (TrkB) at the tumor invasive front as an independent prognostic factor in gastric cancer (2013)
  • Author(s): Koji Tanaka, Tadanobu Shimura, Susumu Saigusa, Satoru Kondo, Takahito Kitajima, Yuji Toiyama, Yoshinaga Okugawa, Yasuhiro Inoue, Masaki Ohi, Toshimitsu Araki, Keiichi Uchida, Yasuhiko Mohri, Masato Kusunoki
  • Organizer: AACR Annual Meeting 2013
  • Place of Presentation: Chicago, Illinois, USA
• [Presentation] Prognostic impact of Vasohibin 1 (VASH1) expression in tumor cells of colorectal cancer patients (2013)
  • Author(s): Takahito Kitajima, Koji Tanaka, Susumu Saigusa, Tadanobu Shimura, Satoru Kondo, Yuji Toiyama, Yoshinaga Okugawa, Yasuhiro Inoue, Masaki Ohi, Toshimitsu Araki, Keiichi Uchida, Yasuhiko Mohri, Hiroki Hori, Masato Kusunoki
  • Organizer: AACR Annual Meeting 2013
  • Place of Presentation: Chicago, Illinois, USA
• [Presentation] Aberrant expression of trefoil factor 3 (TFF3) is associated with tumor progression and an unfavorable prognosis in patients with colorectal cancer (2013)
  • Author(s): Satoru Kondo, Koji Tanaka, Susumu Saigusa, Takahito Kitajima, Tadanobu Shimura, Yuji Toiyama, Yoshinaga Okugawa, Yasuhiro Inoue, Toshimitsu Araki, Keiichi Uchida, Yasuhiko Mohri, Kenichiro Ishii, Masato Kusunoki
  • Organizer: AACR Annual Meeting 2013
  • Place of Presentation: Chicago, Illinois, USA
• [Presentation] Time-series assessment of 5-fluorouracil efficacy on murine colorectal liver metastasis model using intravital multiphoton microscopy (2012)
  • Author(s): Tanaka K et al.
  • Organizer: AACR Annual Meeting 2012
  • Place of Presentation: Chicago, Illinois
• [Presentation] Clinical and molecular comparison between short- and long-course preoperative radiotherapy for rectal cancer (2011)
  • Author(s): Yasuhiro Inoue, Susumu Saigusa, Junichiro Hiro, Yuji Toiyama, Koji Tanaka, Yasuhiko Mohri, Masato Kusunoki
  • Organizer: The 2011 ASCO Annual Meeting
  • Place of Presentation: Chicago, IL
• [Presentation] Macrophage inflammatory protein-3 alpha (MIP-3a) is a novel serum marker for prognosis of colorectal cancer (2010)
  • Author(s): Takashi Iwata, Koji Tanaka, Yasuhiro Inoue, Yuji Toiyama, Junichiro Hiro, Susumu Saigusa, Yasuhiko Mohri, Chikao Miki, Tsutomu Tabata, Masato Kusunoki
  • Organizer: 2010 ASCO Annual Meeting
  • Place of Presentation: CHICAGO, IL
• [Presentation] CD133, OCT4, and NANOG in ulcerative colitis-associated colorectal cancer (2009)
  • Author(s): Tanaka K, Yasuda H, Okita Y, Araki T, Saigusa S, Toiyama Y, Yokoe T, Kawamoto A, Ioue Y, Miki C, Kusunoki M.
  • Organizer: AACR 100th Annual Meeting 2009
  • Place of Presentation: Denver, CO, USA
  • Year and Date: 2009-04-21
• [Book] Oncology Market Research Indicates Standardization of Colorectal Cancer Chemotherapy (2012)
  • Author(s): Yasuhiro Inoue, Koji Tanaka, Masato Kusunoki
  • Publisher: Nova Publishers, New York
• [Book] The Role of Cytoreduction as a Multidisciplinary Treatment Modality for Metastatic Colorectal Cancer. (2010)
  • Author(s): Koji Tanaka, Yasuhiro Inoue, Masato Kusunoki.
  • Publisher: Nova Publishers, New York
• [Book] PTMA (prothymosin, alpha) (2009)
  • Author(s): Tanaka K, Kusunoki M.
  • Publisher: Atlas Genet Cytogenet Oncol Haematol
• [Patent(Industrial Property Rights)] 個体内部の組織学的イメージ像を観察・取得する方法 (2013)
  • Inventor(s): 溝口 明, 山口 輪作
  • Industrial Property Rights Holder: 有限会社 楠屋
  • Acquisition Date: 2013-05-17