Contraction of wild-life iHep cell and development of the evaluation method for metabolism of the top predator

• Project/Area Number: 17K20038
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Environmental analyses and evaluation and related fields
• Research Institution: Hokkaido University
• Principal Investigator: Ikenaka Yoshinori 北海道大学, 獣医学研究院, 准教授 (40543509)
• Co-Investigator(Kenkyū-buntansha): 川合 佑典 帯広畜産大学, 畜産学部, 助教 (10709546) ; 鈴木 淳史 九州大学, 生体防御医学研究所, 教授 (30415195) ; 三谷 曜子 北海道大学, 北方生物圏フィールド科学センター, 准教授 (40538279) ; 石塚 真由美 北海道大学, 獣医学研究院, 教授 (50332474) ; 水川 葉月 愛媛大学, 農学研究科, 准教授 (60612661) ; 中山 翔太 北海道大学, 獣医学研究院, 助教 (90647629)
• Project Period (FY): 2017-06-30 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2018: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2017: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: 野生動物 / iHep / Metabolism / トッププレデター / P450 / UGT / SULT / 化学物質感受性 / 種差 / 誘導性肝臓様細胞（iHep） / 誘導性肝臓様細胞(iHep)

Outline of Final Research Achievements

The purpose of this study is to develop the evaluation method of metabolism, which is the key to chemical sensitivity, and to clarify the chemical metabolic ability of rare top predator. Fresh liver is required for the evaluation of metabolism, but rare species are difficult to obtain. Therefore, in this study, we attempted to differentiate and induce liver-like cells (iHep cells) from skin fibroblasts by direct reprogramming. First, as a result of this study, we succeeded in culturing fibroblasts of several top predators such as brown bear and Sea lion. Furthermore, as a result of attempting to induce iHep cells using cultured fibroblasts, it was found that the cells were transformed into epithelial cells expressing albumin by introducing a specific factor. In this study, it is highly probable that iHep cells were successfully generated starting from wild animals.

Academic Significance and Societal Importance of the Research Achievements

トッププレデターの化学物質感受性を適切に評価できる実験系は無く、その評価法を確立する必要があった。本研究で一部誘導に成功した野生動物iHep細胞は、野生動物の化学物質感受性を非侵襲的かつ迅速に実施できる。更に、iHep細胞は様々な野生動物獣医療にも応用可能である。例えば、投薬や麻酔による不動化の初期スクリーニングに応用する事で、経験則では無く、科学的根拠に基づいた投与doseの決定が可能となる。これにより、獣医療現場における麻酔中の死亡事故などの減少に通じるだけでなく、将来的には各動物種に応じた創薬への応用も期待できる。

Research Products

• [Int'l Joint Research] カセサート大学(タイ)
• [Journal Article] Functional and molecular characterization of UDP-glucuronosyltransferase 2 family in cynomolgus macaques (2019)
  • Author(s): Uno Yasuhiro、Takahira Rika、Murayama Norie、Onozeki Shunsuke、Kawamura Shu、Uehara Shotaro、Ikenaka Yoshinori、Ishizuka Mayumi、Ikushiro Shinichi、Yamazaki Hiroshi
  • Journal Title: Biochemical Pharmacology Volume: 163 Pages: 335-344
  • DOI: 10.1016/j.bcp.2019.03.002
  • Peer Reviewed
• [Journal Article] Characterization of function and genetic feature of UDP-glucuronosyltransferase in avian species (2019)
  • Author(s): Kawai Yusuke K.、Shinya So、Ikenaka Yoshinori、Saengtienchai Aksorn、Kondo Takamitsu、Darwish Wageh Sobhy、Nakayama Shota M.M.、Mizukawa Hazuki、Ishizuka Mayumi
  • Journal Title: Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology Volume: 217 Pages: 5-14
  • DOI: 10.1016/j.cbpc.2018.11.001
  • NAID: 120006803956
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Comparison of xenobiotic metabolism in phase I oxidation and phase II conjugation between rats and bird species (2018)
  • Author(s): Saengtienchai A, Ikenaka Y, Kawata M, Kawai Y, Takeda K, Kondo T, Bortey-Sam N, Nakayama S.M.M, Mizukawa H, Ishizuka M.
  • Journal Title: CBP Volume: 214 Pages: 28-35
  • DOI: 10.1016/j.cbpc.2018.08.007
  • NAID: 120006769940
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Molecular and functional characterization of UDP-glucuronosyltransferase 1A in cynomolgus macaques. (2018)
  • Author(s): Uno, Y., Takahira, R., Murayama, N., Ishii, Y., Ikenaka, Y., Ishizuka, M., Yamazaki, H., and Ikushiro, S.
  • Journal Title: Biochem. Pharmacol. Volume: 155 Pages: 172-181
  • DOI: 10.1016/j.bcp.2018.06.027
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The evolution of UDP-glycosyl/glucuronosyltransferase 1E (UGT1E) genes in bird lineages is linked to feeding habits but UGT2 genes is not. (2018)
  • Author(s): Yusuke K. Kawai, Yoshinori Ikenaka, Mayumi Ishizuka, Akira Kubota
  • Journal Title: PLoS One Volume: 13 Issue: 10 Pages: 1-13
  • DOI: 10.1371/journal.pone.0205266
  • NAID: 120006540774
  • Peer Reviewed / Open Access
• [Journal Article] Comparison of xenobiotic metabolism in phase I oxidation and phase II conjugation between rats and bird species (2018)
  • Author(s): Saengtienchai A, Ikenaka Y, Kawata M, Kawai Y, Takeda K, Kondo T, Bortey-Sam N, Nakayama S.M.M, Mizukawa H, Ishizuka M
  • Journal Title: Comparative Biochemistry & Physiology Part C Volume: in press
  • NAID: 120006769940
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Uridine Diphosphate-Glucuronosyltransferase (UGT) 2B Subfamily Interspecies Differences in Carnivores (2017)
  • Author(s): Kondo Takamitsu、Ikenaka Yoshinori、Nakayama Shouta M. M.、Kawai Yusuke K.、Mizukawa Hazuki、Mitani Yoko、Nomiyama Kei、Tanabe Shinsuke、Ishizuka Mayumi
  • Journal Title: Toxicological Sciences Volume: 158 Issue: 1 Pages: 90-100
  • DOI: 10.1093/toxsci/kfx072
  • Peer Reviewed / Open Access
• [Presentation] 野生哺乳類における硫酸転移酵素の動物種差解明 (2018)
  • Author(s): 近藤誉充、池中良徳、中山翔太、水川葉月、三谷曜子、田辺信介 、野見山桂、石塚真由美
  • Organizer: 第27回環境化学討論会
• [Presentation] 食肉目動物での第II 相抱合酵素の遺伝的性状および酵素学的性状の解明 (2018)
  • Author(s): 近藤誉充、池中良徳、中山翔太、水川葉月、三谷曜子、田辺信介 、野見山桂、石塚真由美
  • Organizer: 第45回 日本毒性学会学術年会
• [Presentation] 鰭脚類を中心とした食肉目の薬物代謝酵素である硫酸転移酵素の種差解明 (2018)
  • Author(s): 近藤 誉充、池中 良徳、中山 翔太、水川 葉月、三谷 曜子、田辺 信介、野見山 桂、石塚 真由美
  • Organizer: 第24回日本野生動物医学会大会
• [Presentation] Inter-species differences of phase II xenobiotics metabolism enzymes in carnivorans. (2018)
  • Author(s): Takamitsu Kondo,Yoshinori Ikenaka, Shouta M.M. Nakayama, Hazuki Mizukawa, Yoko Mitani, Kei Nomiyama, Shinsuke Tanabe, and Mayumi Ishizuka
  • Organizer: The 6th Sapporo Summer Symposium for One Health (6th SaSSOH)
  • Int'l Joint Research
• [Presentation] UDP-glucuronosyltransferase (UGT) among Pinnipedia and other carnivores (2017)
  • Author(s): Takamitsu Kondo, Yoshinori Ikenaka, Shouta M.M. Nakayama, Yusuke K. Kawai, Hazuki Mizukawa, Yoko Mitani, Kei Nomiyama, Shinsuke Tanabe, and Mayumi Ishizuka
  • Organizer: the 19th International Symposium on Pollutant Responses in Marine Organisms (PRIMO 19)
  • Int'l Joint Research
• [Presentation] UDP-glucuronosyltransferase (UGT) 2B genetic feature and activities in Carnivores (2017)
  • Author(s): "Takamitsu Kondo, Yoshinori Ikenaka, Shouta M.M. Nakayama, Yusuke K. Kawai, Hazuki Mizukawa, Yoko Mitani, Kei Nomyama, Shinsuke Tanabe, and Mayumi Ishizuka"
  • Organizer: 9th International Toxicology Symposium in Nigeria
  • Int'l Joint Research
• [Presentation] Drug metabolism enzymes in Carnivora species (2017)
  • Author(s): "Takamitsu Kondo, Yoshinori Ikenaka, Shouta M.M. Nakayama, Yusuke K. Kawai, Hazuki Mizukawa, Yoko Mitani, Kei Nomyama, Shinsuke Tanabe, and Mayumi Ishizuka"
  • Organizer: The 5th Sapporo Summer Seminar for One Health (SaSSOH)
  • Int'l Joint Research