Analysis of oncogene addiction and development of molecular-targeted therapy for small animal tumours

• Project/Area Number: 24380173
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Clinical veterinary science
• Research Institution: Nippon Veterinary and Life Science University
• Principal Investigator: BONKOBARA Makoto 日本獣医生命科学大学, 獣医学部, 准教授 (50343611)
• Co-Investigator(Kenkyū-buntansha): TAZAKI Hiroyuki 日本獣医生命科学大学, 獣医学部, 教授 (80231405) ; OMI Toshinori 日本獣医生命科学大学, 獣医学部, 教授 (40296091) ; TOMIHARI Mizuki 帯広畜産大学, 畜産学部, 講師 (00552754)
• Co-Investigator(Renkei-kenkyūsha): SHIMADA Takashi 日本医科大学, 医学部, 教授 (20125074)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000); Fiscal Year 2014: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2013: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2012: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000)
• Keywords: 犬 / 組織球性肉腫 / oncogene addiction / キナーゼ / ダサチニブ / Oncogene addiction

Outline of Final Research Achievements

To establish foundation of molecular-targeted therapy for small animal tumours, we sought canine tumour-derived cell lines that have “oncogene addiction” and analysed signalling pathway(s) on which they depend. A kinase inhibitor, dasatinib, was found to have potent growth inhibitory activity against some histiocytic sarcoma (HS) lines in vitro and in vivo. We then performed kinome analysis using HS lines and found that 14-3-3 protein gamma is constitutively activated in some HS lines that are sensitive to dasatinib. Because 14-3-3 protein gamma does not have kinase activity, it was considered that dasatinib targeted kinase in upper JNK pathway. From these findings, dasatinib could be a potential therapeutic drug for certain HS cases whose tumour cells have constitutively phosphorylated 14-3-3 protein gamma.

Research Products

• [Journal Article] Molecular changes associated with the development of resistance to imatinib in an imatinib-sensitive canine neoplastic mast cell line carrying a KIT c.1523A>T mutation (2015)
  • Author(s): Kobayashi M, Kuroki S, Tanaka Y, Moriya Y, Kozutumi Y, Uehara Y, Ono K, Tamura K, Washizu T, Bonkobara M.
  • Journal Title: Eur J Haematol. Volume: In press Issue: 6 Pages: 524-31
  • DOI: 10.1111/ejh.12526
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Dysregulation of tyrosine kinases and use of imatinib in small animal practice (2015)
  • Author(s): Bonkobara M.
  • Journal Title: Vet J. Volume: In press Issue: 2 Pages: 180-188
  • DOI: 10.1016/j.tvjl.2014.12.015
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Molecular Cloning and Gene Expression of Canine Apoptosis Inhibitor of Macrophage (2014)
  • Author(s): Tomura S, Uchida M, Yonezawa T, Kobayashi M, Bonkobara M, Arai S, Miyazaki T, Tamahara S, Matsuki N.
  • Journal Title: Journal of Veterinary Medical Science Volume: 76 Issue: 12 Pages: 1641-1645
  • DOI: 10.1292/jvms.14-0166
  • NAID: 130004684872
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access
• [Journal Article] The genetic association study between polymorphisms in uncoupling protein 2 and uncoupling protein 3 and metabolic data in dogs. (2014)
  • Author(s): Udagawa C, Tada N, Asano J, Ishioka K, Ochiai K, Bonkobara M, Tsuchida S, Omi T.
  • Journal Title: BMC Res Notes. Volume: 7 Issue: 1 Pages: 904-904
  • DOI: 10.1186/1756-0500-7-904
  • Peer Reviewed / Open Access
• [Journal Article] Decreased Gene Expressions of Insulin Signal Molecules in Canine Hyperadrenocorticism (2014)
  • Author(s): Nozawa S, Oda H, Akiyama R, Ueda K, Saeki K, Shono S, Maruyama N, Murata A, Tazaki H, Mori A, Momota Y, Azakami D, Sako T, Ishioka K.
  • Journal Title: Journal of Veterinary Medical Science Volume: 76 Issue: 8 Pages: 1177-1182
  • DOI: 10.1292/jvms.14-0033
  • NAID: 130004432193
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] The Silicon Concentration in Cat Urine and Its Relationship with Other Elements (2014)
  • Author(s): Takahashi F, Mochizuki M, Yogo T, Ishioka K, Yumoto N, Sako T, Ueda F, Tagawa M, Tazaki H
  • Journal Title: Journal of Veterinary Medical Science Volume: 76 Issue: 4 Pages: 569-572
  • DOI: 10.1292/jvms.13-0356
  • NAID: 130003382405
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access
• [Journal Article] Low blood cell counts in wild Japanese monkeys after the Fukushima Daiichi nuclear disaster. (2014)
  • Author(s): Ochiai K, Hayama S, Nakiri S, Nakanishi S, Ishii N, Uno T, Kato T, Konno F, Kawamoto Y, Tsuchida S, Omi T.
  • Journal Title: Sci Rep. Volume: 4 Issue: 1 Pages: 5793-5793
  • DOI: 10.1038/srep05793
  • Peer Reviewed / Open Access
• [Journal Article] The proteasome inhibitor bortezomib inhibits the growth of canine malignant melanoma cells in vitro and in vivo. (2013)
  • Author(s): Ito K, Kobayashi M, Kuroki S, Sasaki Y, Iwata T, Mori K, Kuroki T, Ozawa Y, Tetsuka M, Nakagawa T, Hiroi T, Yamamoto H, Ono K, Washizu T, Bonkobara M.
  • Journal Title: Vet J. Volume: ３ Issue: 3 Pages: 577-582
  • DOI: 10.1016/j.tvjl.2013.08.003
• [Journal Article] Analysis of mitochondrial DNA HVR1 haplotype of pure-bred domestic dogs in Japan. (2013)
  • Author(s): Sugiyama S, Chong YH, Shito M, Kasuga M, Kawakami T, Udagawa C, Aoki H, Bonkobara M, Tsuchida S, Sakamoto A, Okuda H, Nagai A, Omi T.
  • Journal Title: Leg Med (Tokyo). Volume: 15 Issue: 6 Pages: 303-309
  • DOI: 10.1016/j.legalmed.2013.08.005
• [Journal Article] Imatinib-associated tumour response in a dog with a non-resectable gastrointestinal stromal tumour harbouring a c-kit exon 11 deletion mutation. (2013)
  • Author(s): Kobayashi M, Kuroki S, Ito K, Yasuda A, Sawada H, Ono K, Washizu T, Bonkobara M.
  • Journal Title: Vet J. Volume: 198 Issue: 1 Pages: 271-274
  • DOI: 10.1016/j.tvjl.2013.05.035
  • Peer Reviewed
• [Journal Article] Molecular cloning and tumour suppressor function analysis of canine REIC/Dkk-3 in mammary gland tumours (2013)
  • Author(s): Ochiai, K., Watanabe, M., Azakami, D., Michishita, M., Yoshikawa, Y., Udagawa, C., Metheenukul, P., Chahomchuen, T., Aoki, H., Kumon, H., Morimatsu, M., Omi, T.
  • Journal Title: Vet. J Volume: 197(3) Issue: 3 Pages: 769-775
  • DOI: 10.1016/j.tvjl.2013.04.024
  • Peer Reviewed
• [Journal Article] Semiquantitative multi-analysis of plasma obtained from Romney lambs (Ovis aries) by inductively coupled plasma mass spectrometry, and the classification according to feed type (2013)
  • Author(s): Takahashi F, Mochizuki M, Sato T, Katayama K, Kenyon PR, Morris ST, Kemp PD, Ozawa, T, Ueda F, Tazaki H
  • Journal Title: Animal Science Journal Volume: in press Issue: 6 Pages: 496-501
  • DOI: 10.1111/asj.12027
  • Peer Reviewed
• [Journal Article] Identification of dasatinib as an in vitro potent growth inhibitor of canine histiocytic sarcoma cells (2013)
  • Author(s): Ito K, Kuroki S, Kobayashi M, Ono K, Washizu T, Bonkobara M
  • Journal Title: Veterinary Journal Volume: (In press) Issue: 3 Pages: 536-540
  • DOI: 10.1016/j.tvjl.2012.12.016
  • Peer Reviewed
• [Journal Article] Validation of the prognostic value of histopathological grading or c-kit mutation in canine cutaneous mast cell tumours: a retrospective cohort study (2013)
  • Author(s): Takeuchi Y, Fujino Y, Watanabe M, Takahashi M, Nakagawa T, Takeuchi A, Bonkobara M, Kobayashi T, Ohno K, Uchida K, Asano K, Nishimura R, Nakayama H, Sugano S, Ohashi Y, Tsujimoto H
  • Journal Title: Vet J Volume: 196 Issue: 3 Pages: 492-498
  • DOI: 10.1016/j.tvjl.2012.11.018
  • Peer Reviewed
• [Journal Article] The DC-HIL ligand syndecan-4 is a negative regulator of T-cell allo-reactivity responsible for graft-versus-host disease. (2013)
  • Author(s): Chung JS, Tomihari M, Tamura K, Kojima T, Cruz PD Jr, Ariizumi K
  • Journal Title: Immunology. Volume: 138(2) Issue: 2 Pages: 173-182
  • DOI: 10.1111/imm.12027
  • Peer Reviewed
• [Journal Article] Non-targeted analyses of animal plasma : betaine and choline represent the nutritional and metabolic status (2012)
  • Author(s): Katayama K, Sato T, Arai T, Amao H, Ohta Y, Ozawa T, Kenyon PR, Tazaki H
  • Journal Title: Journal of Animal Physiology and Animal Nutrition Volume: 97 Issue: 1 Pages: 119-125
  • DOI: 10.1111/j.1439-0396.2011.01250.x
  • Peer Reviewed
• [Journal Article] Investigation of cadmium contamination using hair of the Japanese macaque (Macaca fuscata) from Shimokita Peninsula, Aomori Prefecture in Japan. (2012)
  • Author(s): Mariko MOCHIZUKI
  • Journal Title: Bulltein of Environemtal Contamination and Toxicology Volume: 89 Issue: 3 Pages: 577-579
  • DOI: 10.1007/s00128-012-0694-8
  • Peer Reviewed
• [Journal Article] Establishment of a PCR analysis method for canine BRCA2 (2012)
  • Author(s): Yoshikawa Y, Morimatsu M, Ochiai K, Okuda K, Taoda T, Chikazawa S, Shimamura A, Omi T, Bonkobara M, Orino K, Watanabe K.
  • Journal Title: BMC Res Notes Volume: (in press) Issue: 1 Pages: 173-173
  • DOI: 10.1186/1756-0500-5-173
  • Peer Reviewed
• [Presentation] 獣医療における分子標的薬の将来展望 (2015)
  • Author(s): 盆子原誠
  • Organizer: 獣医内科アカデミー
  • Place of Presentation: 横浜
  • Year and Date: 2015-02-20
  • Invited
• [Presentation] イマチニブの登場と肥満細胞腫の治療 (2014)
  • Author(s): 盆子原誠
  • Organizer: 獣医内科アカデミー
  • Place of Presentation: 横浜
• [Presentation] イマチニブが奏効した犬の消化管間質腫瘍の1症例 (2013)
  • Author(s): 小林正人、黒木詩織、伊藤慶太、安田暁子、澤田治、美小野憲一郎、鷲巣月美、盆子原誠
  • Organizer: 獣医臨床病理学会
  • Place of Presentation: 東京
• [Presentation] Molecular basis of imatinib therapy in mast cell tumors (2013)
  • Author(s): Makoto BONKOBARA
  • Organizer: WSAVA FASAVA 2013
  • Place of Presentation: ニュージーランド
• [Presentation] Molecular-targeting therapy with imatinib in mast cell tumors (2013)
  • Author(s): Makoto BONKOBARA
  • Organizer: WSAVA FASAVA 2013
  • Place of Presentation: ニュージーランド
• [Presentation] In vitro Screening of Compounds led to the Identification of Dasatinib as a Potent Growth Inhibitor for Canine Histiocytic Sarcoma Cells (2013)
  • Author(s): Keita ITO, Shiori KUROKI, Masato KOBAYASHI, Jason Wilson, Tsukimi WASHIZU, Makoto BONKOBARA
  • Organizer: WSAVA FASAVA 2013
  • Place of Presentation: ニュージーランド
• [Presentation] Molecular targeted therapy with imatinib in canine and feline mast cell tumor (2012)
  • Author(s): Makoto BONKOBARA
  • Organizer: 2012 RIVM International Symposium
  • Place of Presentation: 韓国
• [Presentation] Development of Y-chromosomal microsatellite markers in Japanese macaque (Macaca fuscata) population of Fukushima (2012)
  • Author(s): Toshinori OMI, Masayuki SHITOH, Chihiro UDAGAWA, Naomi DADA, Yong Hwa CHONG, Kazuhiko OCHIAI, Atushi SAKAMOTO, Shuichi TSUCHIDA, Shinichi HAYAMA
  • Organizer: 33th International Conference on Animal Genetics
  • Place of Presentation: オーストラリア
• [Presentation] 犬組織球性肉腫に対するダサチニブの増殖抑制効果に関する研究 (2012)
  • Author(s): 伊藤慶太、池谷早代、竹下恭平、黒木詩織、小林正人、盆子原誠、鷲巣月美
  • Organizer: 第154回日本獣医学会学術集会
  • Place of Presentation: 岩手
• [Presentation] 犬の悪性黒色腫細胞に対するBortezomibの増殖抑制効果に関する研究 (2012)
  • Author(s): 手塚雅典、中川朋子、廣井輝代、山本仁美、小澤佑太、伊藤慶太、黒木詩織、小林正人、盆子原誠、鷲巣月美
  • Organizer: 第154回日本獣医学会学術集会
  • Place of Presentation: 岩手
• [Presentation] 犬のイマチニブ耐性肥満細胞腫細胞株の樹立と耐性化機構の解析 (2012)
  • Author(s): 田中優、小堤友香梨、守谷友加里、上原裕司、小林正人、杉崎修、伊藤慶太、黒木詩織、盆子原誠、鷲巣月美
  • Organizer: 第154回日本獣医学会学術集会
  • Place of Presentation: 岩手
• [Presentation] イヌ乳腺腫瘍関連遺伝子BRCA2のBRC repeat 3に存在するSNPsの頻度および機能に及ぼす影響の検討 (2012)
  • Author(s): 落合和彦、塩入弓絵、染田沙織、大澤有加、宇田川智野、森松正美、吉川泰永、盆子原誠、土田修一、近江俊徳
  • Organizer: 第155回日本獣医学会学術集会
  • Place of Presentation: 東京
• [Presentation] イヌUCP2およびUCP3 : SNPディスカバリーと2犬種のSNPジェノタイピング (2012)
  • Author(s): 宇田川智野、多田尚美、青木博史、盆子原誠、土田修一、落合和彦、石岡克己、鄭英和、近江俊徳
  • Organizer: 第155回日本獣医学会学術集会
  • Place of Presentation: 東京
• [Remarks] 日本獣医生命科学大学 科学研究費補助金
  • URL: http://www.nvlu.ac.jp/research/005.html/