Establishment of smart-diagnosis&treatment for canine tumor by exosome

• Project/Area Number: 17H03926
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: Kagoshima University
• Principal Investigator: Miura Naoki 鹿児島大学, 農水産獣医学域獣医学系, 教授 (80508036)
• Co-Investigator(Kenkyū-buntansha): 川原 幸一 大阪工業大学, 工学部, 教授 (10381170) ; 福島 隆治 東京農工大学, (連合)農学研究科(研究院), 教授 (10466922) ; 丸山 征郎 鹿児島大学, 医歯学総合研究科, 特任教授 (20082282) ; 中川 貴之 東京大学, 大学院農学生命科学研究科(農学部), 准教授 (40447363) ; 畑井 仁 鹿児島大学, 農水産獣医学域獣医学系, 准教授 (40566535) ; 岡本 芳晴 鳥取大学, 農学部, 教授 (50194410) ; 後藤 章暢 兵庫医科大学, 医学部, 教授 (70283885) ; 原田 陽一郎 地方独立行政法人大阪府立病院機構大阪国際がんセンター(研究所), その他部局等, 糖鎖オンコロジー部主任研究員 (80464147)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000); Fiscal Year 2019: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2018: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2017: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
• Keywords: ノンコーディングRNA / マイクロRNA / エクソソーム / 腫瘍 / 犬 / メラノーマ / 肝細胞癌 / 乳腺腫瘍 / イヌ / 犬腫瘍 / 診断・治療 / non-coding RNA / イヌ腫瘍 / エキソソーム / small RNA

Outline of Final Research Achievements

We studied the small-non-coding RNA and mRNA that change specifically in canine tumors to development for new diagnosis and treatment strategy. We also analyzed the tumor-specific exosome as well. As a result, we discovered RNA molecules that cause specific alternative expression in several different canine tumors using the next-generation sequence technology. We also identified the deferentially expressed small-non-coding RNA in the exosome from melanoma. This deferentially expressed pattern depends on the melanoma development and metastasis condition. Finally, we also found the similarities of RNA molecule expression pattern between humans and dogs with our transcriptome data.

Academic Significance and Societal Importance of the Research Achievements

腫瘍に特異的な変化を示すsmall-non-coding RNAは、人医療でも獣医医療でも、液体バイオプシー（Liquid Biopsy）として、早期の的確な診断を達成するために重要なテーマである。本研究の成果である犬の腫瘍（エクソソーム）特異的なこれらの分子の発見は、直接、獣医医療の発展に貢献する。さらに、人と犬の腫瘍の類似性をRNAレベルで比較することで、本研究の成果は獣医医療のみならず、人医療にも応用発展できる可能性を示した。

Research Products

• [Journal Article] Aberrantly expressed snoRNA, snRNA, piRNA and tRFs in canine melanoma (2019)
  • Author(s): Rahman Md. Mahfuzur、Lai Yu-Chang、Husna Al Asmaul、Chen Hui-Wen、Tanaka Yuiko、Kawaguchi Hiroaki、Hatai Hitoshi、Miyoshi Noriaki、Nakagawa Takayuki、Fukushima Ryuji、Miura Naoki
  • Journal Title: Veterinary and Comparative Oncology Volume: - Issue: 3 Pages: 353-361
  • DOI: 10.1111/vco.12558
  • Peer Reviewed / Open Access
• [Journal Article] Transcriptome analysis of dog oral melanoma and its oncogenic analogy with human melanoma (2019)
  • Author(s): Rahman Md. Mahfuzur、Lai Yu-Chang、Husna Al、Chen Hui-Wen、Tanaka Yuiko、Kawaguchi Hiroaki、Hatai Hitoshi、Miyoshi Noriaki、Nakagawa Takayuki、Fukushima Ryuji、Miura Naoki
  • Journal Title: Oncology Reports Volume: 43 Pages: 16-30
  • DOI: 10.3892/or.2019.7391
  • Peer Reviewed / Open Access
• [Journal Article] Micro RNA Transcriptome Profile in Canine Oral Melanoma (2019)
  • Author(s): Rahman Md. Mahfuzur、Lai Yu-Chang、Husna Al Asmaul、Chen Hui-wen、Tanaka Yuiko、Kawaguchi Hiroaki、Miyoshi Noriaki、Nakagawa Takayuki、Fukushima Ryuji、Miura Naoki
  • Journal Title: International Journal of Molecular Sciences Volume: 20 Issue: 19 Pages: 4832-4832
  • DOI: 10.3390/ijms20194832
  • Peer Reviewed / Open Access
• [Journal Article] Bovine milk transcriptome analysis reveals microRNAs and RNU2 involved in mastitis (2019)
  • Author(s): Lai Yu-Chang、Lai Yu-Ting、Rahman Md Mahfuzur、Chen Hui-Wen、Husna Al Asmaul、Fujikawa Takuro、Ando Takaaki、Kitahara Go、Koiwa Masateru、Kubota Chikara、Miura Naoki
  • Journal Title: The FEBS Journal Volume: 287 Issue: 9 Pages: 1899-1918
  • DOI: 10.1111/febs.15114
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] N‐glycome inheritance from cells to extracellular vesicles in B16 melanomas (2019)
  • Author(s): Harada Yoichiro、Kizuka Yasuhiko、Tokoro Yuko、Kondo Kiyotaka、Yagi Hirokazu、Kato Koichi、Inoue Hiromasa、Taniguchi Naoyuki、Maruyama Ikuro
  • Journal Title: FEBS Letters Volume: 印刷中 Issue: 9 Pages: 942-951
  • DOI: 10.1002/1873-3468.13377
  • Peer Reviewed / Open Access
• [Journal Article] Application of high-mannose-type glycan-specific lectin from Oscillatoria Agardhii for affinity isolation of tumor-derived extracellular vesicles (2019)
  • Author(s): Yamamoto Mika、Harada Yoichiro、Suzuki Takehiro、Fukushige Tomoko、Yamakuchi Munekazu、Kanekura Takuro、Dohmae Naoshi、Hori Kanji、Maruyama Ikuro
  • Journal Title: Analytical Biochemistry Volume: 580 Pages: 21-29
  • DOI: 10.1016/j.ab.2019.06.001
  • Peer Reviewed
• [Journal Article] Generation of the heterogeneity of extracellular vesicles by membrane organization and sorting machineries (2019)
  • Author(s): Harada Yoichiro、Suzuki Takehiro、Fukushige Tomoko、Kizuka Yasuhiko、Yagi Hirokazu、Yamamoto Mika、Kondo Kiyotaka、Inoue Hiromasa、Kato Koichi、Taniguchi Naoyuki、Kanekura Takuro、Dohmae Naoshi、Maruyama Ikuro
  • Journal Title: Biochimica et Biophysica Acta (BBA) - General Subjects Volume: 1863 Issue: 4 Pages: 681-691
  • DOI: 10.1016/j.bbagen.2019.01.015
  • Peer Reviewed
• [Journal Article] Comparison between blood coagulability in the intra-atrial and peripheral regions during the acute phase after rapid atrial pacing (2019)
  • Author(s): Yamada S, Hirao D, Miura N, Iwanaga T, Kawaguchi T, Yoshimura A, Oomori T, Nagasato T, Maruyama I, Fukushima R
  • Journal Title: Experimental Animals Volume: 68 Issue: 2 Pages: 137-146
  • DOI: 10.1538/expanim.18-0100
  • NAID: 130007642989
  • ISSN: 0007-5124, 1341-1357, 1881-7122
  • Peer Reviewed / Open Access
• [Journal Article] DNA aneuploidy and centrosome amplification in canine tumor cell lines (2019)
  • Author(s): Endo Yoshifumi、Watanabe Manabu、Miyajima-Magara Nozomi、Igarashi Maki、Mochizuki Manabu、Nishimura Ryohei、Sugano Sumio、Sasaki Nobuo、Nakagawa Takayuki
  • Journal Title: Tissue and Cell Volume: 61 Pages: 67-71
  • DOI: 10.1016/j.tice.2019.09.003
  • Peer Reviewed
• [Journal Article] Anti-tumor effects of the histone deacetylase inhibitor vorinostat on canine urothelial carcinoma cells (2019)
  • Author(s): Eto Shotaro、Saeki Kohei、Yoshitake Ryohei、Yoshimoto Sho、Shinada Masahiro、Ikeda Namiko、Kamoto Satoshi、Tanaka Yuiko、Kato Daiki、Maeda Shingo、Tsuboi Masaya、Chambers James、Uchida Kazuyuki、Nishimura Ryohei、Nakagawa Takayuki
  • Journal Title: PLOS ONE Volume: 14 Issue: 6 Pages: 0218382-0218382
  • DOI: 10.1371/journal.pone.0218382
  • Peer Reviewed / Open Access
• [Journal Article] A Basic Study of Photodynamic Therapy with Glucose-Conjugated Chlorin e6 Using Mammary Carcinoma Xenografts (2019)
  • Author(s): Osaki Tomohiro、Hibino Shota、Yokoe Inoru、Yamaguchi Hiroaki、Nomoto Akihiro、Yano Shigenobu、Mikata Yuji、Tanaka Mamoru、Kataoka Hiromi、Okamoto Yoshiharu
  • Journal Title: Cancers Volume: 11 Issue: 5 Pages: 636-636
  • DOI: 10.3390/cancers11050636
  • Peer Reviewed / Open Access
• [Journal Article] Antimalarial Drugs Enhance the Cytotoxicity of 5-Aminolevulinic Acid-Based Photodynamic Therapy against the Mammary Tumor Cells of Mice In Vitro (2019)
  • Author(s): Osaki Tomohiro、Takahashi Kiwamu、Ishizuka Masahiro、Tanaka Tohru、Okamoto Yoshiharu
  • Journal Title: Molecules Volume: 24 Issue: 21 Pages: 3891-3891
  • DOI: 10.3390/molecules24213891
  • Peer Reviewed
• [Journal Article] Lentiviral vector-mediated transfection of p53, p16 and PTEN genes against human renal cell carcinoma cell lines. (2019)
  • Author(s): Wataru Matsunaga, Misa Ichikawa, Takahiro Ishikawa, Akinobu Gotoh.
  • Journal Title: Personalized Medicine Universe. Volume: 8 Pages: 10-14
  • Peer Reviewed
• [Journal Article] Lentiviral vector-mediated gene transfer combined with cisplatin enhances tumor suppression in human bladder cancer cell lines. (2019)
  • Author(s): Misa Ichikawa, Wataru Matsunaga, Takahiro Ishikawa, Akinobu Gotoh.
  • Journal Title: Personalized Medicine Universe Volume: 8 Pages: 15-19
  • Peer Reviewed
• [Journal Article] Identification of dysregulated microRNAs in canine malignant melanoma. (2019)
  • Author(s): Ushio N, Rahman MM, Maemura T, Lai YC, Iwanaga T, Kawaguchi H, Miyoshi N, Momoi Y, Miura N.
  • Journal Title: Oncolgy Letter Volume: 17 Pages: 1080-1088
  • DOI: 10.3892/ol.2018.9692
  • Peer Reviewed / Open Access
• [Journal Article] Aberrant expression of microRNAs and the miR-1/MET pathway in canine hepatocellular carcinoma (2018)
  • Author(s): Lai YC, Ushio N, Rahman MM, Katanoda Y, Ogihara K, Naya Y, Moriyama A, Iwanaga T, Saitoh Y, Sogawa T, Sunaga T, Momoi Y, Izumi H, Miyoshi N, Endo Y, Fujiki M, Kawaguchi H, Miura N.
  • Journal Title: Vet Comp Oncol. Volume: in press Issue: 2 Pages: 0-0
  • DOI: 10.1111/vco.12379
  • Peer Reviewed / Open Access
• [Journal Article] Rapid Communication: MiR-92a as a housekeeping gene for analysis of bovine mastitis-related microRNA in milk. (2017)
  • Author(s): Lai YC, Fujikawa T, Ando T, Kitahara G, Koiwa M, Kubota C, Miura N
  • Journal Title: J Anim Sci Volume: 95 Issue: 6 Pages: 2732-2735
  • DOI: 10.2527/jas.2017.1384
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Effects of toceranib phosphate (Palladia) monotherapy on multidrug resistant lymphoma in dogs (2017)
  • Author(s): Yamazaki H, Miura N, Lai YC, Takahashi M, Goto-Koshino Y, Yasuyuki M, Nakaichi M, Tsujimoto H, Setoguchi A, Endo Y.
  • Journal Title: Journal of Veterinary Medical Science Volume: 79 Issue: 7 Pages: 1225-1229
  • DOI: 10.1292/jvms.16-0457
  • NAID: 130005808374
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Inflammation-related microRNA expression level in the bovine milk is affected by mastitis. (2017)
  • Author(s): Lai YC, Fujikawa T, Maemura T, Ando T, Kitahara G, Endo Y, Yamato O, Koiwa M, Kubota C, Miura N
  • Journal Title: PLoS One Volume: 12(5) Issue: 5 Pages: 0177182-0177182
  • DOI: 10.1371/journal.pone.0177182
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Hypoxia-activated prodrug TH-302 decreased survival rate of canine lymphoma cells under hypoxic condition (2017)
  • Author(s): Hiroki Yamazakia, Yu-Chang Laia, Morihiro Tatenob, Asuka Setoguchic, Yuko Goto-Koshinod, Yasuyuki Endob, Munekazu Nakaichie, Hajime Tsujimotod, Naoki Miura
  • Journal Title: PLOS ONE Volume: 印刷中 Issue: 5 Pages: 0177305-0177305
  • DOI: 10.1371/journal.pone.0177305
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] The application of small non-coding RNA in the veterinary field. Primarily focus the transcriptome analysis in canine tumor and bovine mastitis. (2020)
  • Author(s): Miura N
  • Organizer: ASCon XXVI, BSVER
  • Invited
• [Presentation] Transcriptome analysis of canine oral melanoma and its human analogy (2019)
  • Author(s): Rahman R, Lai YC, Husna A, Chen HW, Nakagawa T, Fukushima R, Miura N
  • Organizer: 9th Mildred Scheel cancer conference
  • Int'l Joint Research
• [Presentation] Evaluation of the Total-Thrombus Analysis System (T-TAS) in dogs at risk of bleeding (2019)
  • Author(s): Iwanaga T, Miura N, Brainard B, Brooks MB, Goggs R
  • Organizer: IVECCS
  • Int'l Joint Research
• [Presentation] Lentiviral vector mediated transfection of tumor suppressor genes. (2019)
  • Author(s): Matsunaga, W. Ichikawa, M. Ishikawa, T. Gotoh, A.
  • Organizer: 第25回日本遺伝子細胞治療学会
• [Presentation] Combined effect of Lentiviral vector mediated gene transfer and cisplatin. (2019)
  • Author(s): Ichikawa, M. Matsunaga, W. Ishikawa, T. Gotoh, A.
  • Organizer: 第25回日本遺伝子細胞治療学会
• [Presentation] TNF-a induction by Histone H2A and H2B is mediated via ERK1/2 in RAW264.7 cells. (2019)
  • Author(s): Ueno H, Nakatomi K, Anan S, Tancharoen S, Kikuchi K, Ito T, Maruyama I, Kawahara KI
  • Organizer: 9th International DAMPs and Alarmins Symposium
  • Int'l Joint Research
• [Presentation] NSAIDs inhibit IDO1 gene expression in canine transitional cell carcinoma cell lines. (2019)
  • Author(s): Ikeda N, Kato D, EtoS, Yoshitake R, Yoshimoto S, Shinada M, Kamoto S, Fujita N, Nishimura R, Nakagawa T.
  • Organizer: The 6th Asian Meeting of Animal Medicine Specialties, Shanghai, China(Shanghai Convention & Exhibition Center of International Sourcing)
  • Int'l Joint Research
• [Presentation] Sugar-linked chlorin e6 for photodynamic therapy (2019)
  • Author(s): Yano S, Kataoka H, Nishie H, Tanaka M, Osaki T, Okamoto Y, Nomoto A.
  • Organizer: The 26th International SPACC Symposium
  • Int'l Joint Research
• [Presentation] microRNA-mRNA transcriptomic crosstalk of canine melanoma (2017)
  • Author(s): Rahman MM, Lai YC, Ushio N, Tanaka Y, Hatai H, Nakagawa T, Miyoshi N, Miura N.
  • Organizer: ConBio2017(第40回日本分子生物学会年会）