Monitoring of transplanted hepatocytes and analysis of immune rejection responses in the hepatocyte chimeric mouse system.

• Project/Area Number: 16K09354
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Gastroenterology
• Research Institution: Nagoya University
• Principal Investigator: Ishikawa Tetsuya 名古屋大学, 医学系研究科(保健), 教授 (10288508)
• Co-Investigator(Kenkyū-buntansha): 伊藤 弘康 岐阜大学, 大学院医学系研究科, 准教授 (80373075)
• Research Collaborator: YUKAWA Hiroshi ; MATSUNAGA Tamihide ; SUEMIZU Hiroshi ; HAYASHI Yumi
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: 肝細胞キメラマウス / 細胞移植 / 免疫拒絶 / 肝不全 / 肝細胞移植 / 肝細胞置換率 / GFP / 戻し交配 / 量子ドット / 再生医療 / イメージング / GVHD

Outline of Final Research Achievements

Hepatocyte-chimera mouse system was established as a model for the cell transplantation treatment with hepatocytes or stem cell-derived cells. In the transgenic mice, HSVtk that express the thymidine kinase of human herpes simplex virus in their hepatocytes, administration of ganciclovir (GCV) kills autologous hepatocytes, enabling transplantation of hepatocytes of other origin. In the case of allogenic hepatocyte transplantation into HSVtk, all the transplanted cells were eliminated by a strong immune-rejection response. In the case of syngeneic hepatocytes expressing foreign antigens, efficient engraftment of transplanted cells was observed, while inflammatory cells infiltration around the engrafted cells and cellular immune response to the foreign antigens were confirmed. The hepatocyte-chimeric mouse system can be a useful model for elucidating the mechanism of immune rejection in the cell transplantation in the liver and for studying a therapeutic method for immune rejection.

Academic Significance and Societal Importance of the Research Achievements

重症な急性・慢性肝不全に対する治療として、生体肝移植、脳死肝移植が実施されているものの、ドナーの絶対的な不足により、適応症例に対しても治療の提供が困難な状況が続いている。肝細胞あるいは幹細胞由来肝様細胞の移植治療はこの状況を改善し得る有力な治療法と考えられるが、その有用性を検証できる動物モデルは確立されていない。本研究で用いた肝細胞キメラマウス作製系は、そのモデルとなり得るとともに、他家由来肝細胞移植に伴う免疫拒絶の病態解明、それに対する治療法確立のための有用なモデルともなり得る。

Research Products

• [Journal Article] In utero exposure to di(2-ethylhexyl)phthalate suppresses blood glucose and leptin levels in the offspring of wild-type mice. (2019)
  • Author(s): Hayashi Y, Ito Y, Naito H, Tamada H, Yamagishi N, Kondo T, Ishikawa T, Gonzalez FJ, Nakajima T.
  • Journal Title: Toxicology Volume: 415 Pages: 49-55
  • DOI: 10.1016/j.tox.2019.01.008
  • Peer Reviewed
• [Journal Article] In Vivo Real-Time Monitoring System Using Probe Electrospray Ionization/Tandem Mass Spectrometry for Metabolites in Mouse Brain (2018)
  • Author(s): Zaitsu Kei、Hayashi Yumi、Murata Tasuku、Yokota Kazumi、Ohara Tomomi、Kusano Maiko、Tsuchihashi Hitoshi、Ishikawa Tetsuya、Ishii Akira、Ogata Koretsugu、Tanihata Hiroshi
  • Journal Title: Analytical Chemistry Volume: 90 Issue: 7 Pages: 4695-4701
  • DOI: 10.1021/acs.analchem.7b05291
  • Peer Reviewed
• [Journal Article] Metabolome analysisof the serotonin syndrome rat model: abnormal muscular contraction is relatedto metabolic alterations and hyper-thermogenesis (2018)
  • Author(s): Kei Zaitsu*, Saki Noda, Akira Iguchi, Yumi Hayashi, Tomomi Ohara, YuyaKimura, Yuta Koketsu, Tomoko Kosaki, Maiko Kusano, Takako Sato, TetsuyaIshikawa, Hitoshi Tsuchihashi, Koichi Suzuki, Akira Ishii.
  • Journal Title: Life Sciences Volume: 207 Pages: 550-561
  • DOI: 10.1016/j.lfs.2018.06.031
  • Peer Reviewed
• [Journal Article] Imaging of angiogenesis of human umbilical vein endothelial cells by uptake of exosomes secreted from hepatocellular carcinoma cells. (2018)
  • Author(s): Yukawa H, Suzuki K, Aoki K, Arimoto T, Yasui T, Kaji N, Ishikawa T, Ochiya T, Baba Y.
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 6765-6765
  • DOI: 10.1038/s41598-018-24563-0
  • Peer Reviewed / Open Access
• [Journal Article] Effective Transplantation of 2D and 3D Cultured Hepatocyte Spheroids Confirmed by Quantum Dot Imaging (2018)
  • Author(s): Okumura H., Nanizawa E., Nakanishi A., Yukawa H., Hashita T., Iwao T., Baba Y., Ishikawa T., Matsunaga T.
  • Journal Title: Advanced Biosystems Volume: 2 Issue: 8 Pages: 1800137-1800137
  • DOI: 10.1002/adbi.201800137
  • Peer Reviewed / Open Access
• [Journal Article] Secreted ectodomain of SIGLEC-9 and MCP-1 synergistically improve acute liver failure in rats by altering macrophage polarity (2017)
  • Author(s): Ito T, Ishigami M, Matsushita Y, Hirata M, Matsubara K, Ishikawa T, Hibi H, Ueda M, Hirooka Y, Goto H, Yamamoto A
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 44043-44043
  • DOI: 10.1038/srep44043
  • NAID: 120006306638
  • Peer Reviewed / Open Access
• [Journal Article] Intact metabolite profiling of mouse brain by probe electrospray ionization/triple quadrupole tandem mass spectrometry (PESI/MS/MS) and its potential use for local distribution analysis of the brain (2017)
  • Author(s): Hayashi Yumi、Zaitsu Kei、Murata Tasuku、Ohara Tomomi、Moreau St?phane、Kusano Maiko、Tanihata Hiroshi、Tsuchihashi Hitoshi、Ishii Akira、Ishikawa Tetsuya
  • Journal Title: Analytica Chimica Acta Volume: 983 Pages: 160-165
  • DOI: 10.1016/j.aca.2017.06.047
  • Peer Reviewed
• [Journal Article] Hepatocyte-specific depletion of ubiquitin regulatory X domain containing protein 8 accelerates fibrosis in a mouse non-alcoholic steatohepatitis model (2017)
  • Author(s): Imai Norihiro、Suzuki Michitaka、Ishizu Yoji、Kuzuya Teiji、Honda Takashi、Hayashi Kazuhiko、Ishigami Masatoshi、Hirooka Yoshiki、Ishikawa Tetsuya、Goto Hidemi、Fujimoto Toyoshi
  • Journal Title: Histochemistry and Cell Biology Volume: 148 Issue: 3 Pages: 219-227
  • DOI: 10.1007/s00418-017-1572-6
  • Peer Reviewed
• [Journal Article] (-)-Epigallocatechin-3-gallate Down-regulates Doxorubicin-induced Overexpression of P-glycoprotein Through the Coordinate Inhibition of PI3K/Akt and MEK/ERK Signaling Pathways. (2017)
  • Author(s): Satonaka H, Ishida K, Takai M, Koide R, Shigemasa R, Ueyama J, Ishikawa T, Hayashi K, Goto H, Wakusawa S.
  • Journal Title: Anticancer Res Volume: 37 Pages: 6071-6077
  • Peer Reviewed
• [Journal Article] Effects of ursodeoxycholic acid and insulin on palmitate-induced ROS production and down-regulation of PI3K/Akt signaling activity. (2017)
  • Author(s): Yokoyama K, Tatsumi Y, Hayashi K, Goto H, Ishikawa T, Wakusawa S.
  • Journal Title: Biol Pharm Bull Volume: 40 Pages: 2001-2004
  • NAID: 130006191810
• [Presentation] 量子ドットイメージング技術を用いた移植幹細胞と免疫細胞のinteraction 機構解明 (2018)
  • Author(s): 北村晃大、湯川博、佐藤和秀、有本知子、小野島大介、石川哲也、馬場嘉信
  • Organizer: 第39回日本炎症・再生医学会
• [Presentation] iPS細胞由来肝様細胞に対する量子ドットを用いたラベル化手法の確立と移植後のin vivoイメージング (2018)
  • Author(s): 名仁澤英里、林由美、湯川博、石川哲也
  • Organizer: 第39回日本炎症・再生医学会
• [Presentation] 探針エレクトロスプレーイオン化タンデム質量分析（PESI/MS/MS）とScheduled SRM法を用いた生体組織のインタクト・メタボロミクス法の構築 (2018)
  • Author(s): 近藤健太、林由美、村田匡、土屋弥月、大原倫美、石川哲也、緒方是嗣、土橋均、石井晃、財津桂
  • Organizer: 第43回日本医用マススペクトル学会年会
• [Presentation] 探針エレクトロスプレーイオン化タンデム質量分析（PESI/MS/MS）を用いたin vivoリアルタイム・モニタリングシステムの構築 (2018)
  • Author(s): 林由美、財津桂、村田匡、土橋均，石井晃、緒方是嗣、石川哲也
  • Organizer: 第43回日本医用マススペクトル学会年会
• [Presentation] マウス肝障害モデルにおけるトロンボモジュリン遺伝子導入脂肪由来肝細胞の炎症軽減効果について (2018)
  • Author(s): 山本万智、名仁澤英里、今井田藍、山崎花那子、園玲華、玉置優貴、宮下凛太郎、林由美、石川哲也
  • Organizer: 第65回日本臨床検査医学会学術集会
• [Presentation] 量子ドットイメージング技術を用いた移植幹細胞と免疫細胞のinteraction 機構解明 (2018)
  • Author(s): 湯川 博、佐藤 和秀、有本 知子、小野島 大介、石川 哲也、馬場 嘉信
  • Organizer: 第17回日本再生医療学会総会
• [Presentation] 歯髄幹細胞無血清培養上清による難治性肝疾患での有用性の検討-急性・慢性肝炎動物モデルにおける効果発現メカニズムの違いに着目して (2017)
  • Author(s): 伊藤隆徳、田中卓、山本健太、安藤祐資、安田諭、野村彩、加藤幸一郎、石津洋二、葛谷貞二、本多隆、林和彦、石上雅敏、廣岡芳樹、石川哲也、後藤秀実
  • Organizer: 第53回日本肝臓学会総会
• [Presentation] マウスモデルを使用した慢性B型肝炎に対する治療用ワクチン開発の試み (2017)
  • Author(s): 伊藤弘康、清島満、石川哲也
  • Organizer: 第53回日本肝臓学会総会
• [Presentation] Rapid-fire direct metabolome analysis by probe electrospray ionization/tandem mass spectrometry: its preclinical applications to validate the methods. (2017)
  • Author(s): Hayashi Y, Zaitsu K, Murata T, Ohara T, Kondo K, Kusano M, Ogata K, Tsuchihashi H, Tanihata H, Ishii A, Ishikawa T.
  • Organizer: The 4th Annual European Congress of the Association for Mass Spectrometry
• [Book] B型肝炎ワクチンとアジュバント「次世代アジュバント開発のためのメカニズム解明と安全性評価」 (2017)
  • Author(s): 伊藤弘康、石川哲也
  • Publisher: ファインケミカル
  • ISBN: 9784781312385
• [Book] 血液検査：免疫学的検査、一般検査「ここまできた肝臓病診療」 (2017)
  • Author(s): 石川哲也
  • Publisher: 中山書店