Studies on intracellular and extracellular pathways involved in drug-induced pulmonary fibrosis

• Project/Area Number: 18K06749
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 47060:Clinical pharmacy-related
• Research Institution: Hiroshima University
• Principal Investigator: Yumoto Ryoko 広島大学, 医系科学研究科(薬), 准教授 (70379915)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 薬剤誘発性肺線維症 / 上皮間葉転換(EMT) / 肺胞上皮Ⅱ型細胞 / 細胞周期の停止 / 細胞外分泌因子(PAI-1) / 肺線維症モデル動物 / 薬剤性肺線維症 / 上皮間葉転換 / 細胞外分泌因子 / PAI-1 / 肺胞上皮細胞 / ERストレス / ATF3/4 / 薬学 / 薬剤性肺障害 / 細胞周期停止 / サイトカイン / TGF-β1 / 上皮間葉転換 (EMT) / 細胞内経路

Outline of Final Research Achievements

Several anticancer drugs including methotrexate (MTX) cause serious lung diseases such as pulmonary fibrosis. Although evidences showing the association of epithelial-mesenchymal transition (EMT) with pulmonary fibrosis are increasing, the mechanism underlying anticancer drug-induced EMT has been poorly understood.
In this research, we indicated that MTX-induced EMT may be related to cell cycle arrest in human alveolar epithelial cell line, A549. It was also suggested that the transforming growth factor (TGF)-β-related signaling pathway may partly be involved in MTX-induced EMT, and MTX may induce EMT via upregulation of ATF3 expression and increase of PAI-1 secretion followed by interaction of PAI-1 with uPAR

Academic Significance and Societal Importance of the Research Achievements

薬剤性肺障害のうち、肺線維症は非常に重篤な障害であるが、その発症機構を細胞・分子レベルで解析した報告は乏しく、また有効な予防方法・治療方法はない。
本研究では薬剤誘発性EMTによる肺線維症に関わる分子機構を細胞内経路、細胞外経路の両面から解明することによって、肺線維症に対する新たな予防法や治療戦略を考案し得るところに学術的・社会的意義があると考える。

Research Products

• [Journal Article] Role of Plasminogen Activator inhibitor-1 in Methotrexate-Induced Epithelial-Mesenchymal Transition in Alveolar Epithelial A549 Cells (2020)
  • Author(s): Yamagami Y, Kawami M, Ojima T, Futatsugi S, Yumoto R, Takano M
  • Journal Title: Biochem Biophys Res Commun Volume: 525 Issue: 3 Pages: 543-548
  • DOI: 10.1016/j.bbrc.2020.02.131
  • Peer Reviewed
• [Journal Article] Effect of transforming growth factor-β1 on functional expression of monocarboxylate transporter 1 in alveolar epithelial A549 cells. (2020)
  • Author(s): Uddin, M., Kawami, M., Yumoto, R., Takano, M.
  • Journal Title: Naunyn Schmiedebergs Arch Pharmacol. Volume: 393 Issue: 5 Pages: 889-896
  • DOI: 10.1007/s00210-019-01802-3
  • Peer Reviewed
• [Journal Article] miR-484: A Possible Indicator of Drug-Induced Pulmonary Fibrosis (2020)
  • Author(s): Konaka Takashi、Kawami Masashi、Yamamoto Ayano、Yumoto Ryoko、Takano Mikihisa
  • Journal Title: Journal of Pharmacy & Pharmaceutical Sciences Volume: 23 Pages: 486-495
  • DOI: 10.18433/jpps31448
  • Peer Reviewed
• [Journal Article] Differential mechanisms underlying methotrexate-induced cell death and epithelial-mesenchymal transition in A549 cells (2020)
  • Author(s): Ojima Takamichi、Kawami Masashi、Yumoto Ryoko、Takano Mikihisa
  • Journal Title: Toxicological Research Volume: - Issue: 3 Pages: 293-300
  • DOI: 10.1007/s43188-020-00067-w
  • Peer Reviewed
• [Journal Article] Suppressive effect of quercetin against bleomycin-induced epithelial-mesenchymal transition in alveolar epithelial cells (2020)
  • Author(s): Takano Mikihisa、Deguchi Junya、Senoo Shunsuke、Izumi Miho、Kawami Masashi、Yumoto Ryoko
  • Journal Title: Drug Metabolism and Pharmacokinetics Volume: 35 Issue: 6 Pages: 522-526
  • DOI: 10.1016/j.dmpk.2020.08.001
  • NAID: 50014656611
  • Peer Reviewed
• [Journal Article] Investigation on inhibitory effect of folic acid on methotrexate-induced epithelial-mesenchymal transition focusing on dihydrofolate reductase. (2019)
  • Author(s): Kawami, M., Honda, N., Hara, T., Yumoto, R. and Takano, M.
  • Journal Title: Drug Metab. Pharmacokinet. Volume: 34 Issue: 6 Pages: 396-399
  • DOI: 10.1016/j.dmpk.2019.08.003
  • NAID: 50014558530
  • Peer Reviewed
• [Journal Article] Association of cell cycle arrest with anticancer drug-induced epithelial-mesenchymal transition in alveolar epithelial cells. (2019)
  • Author(s): Kawami, M., Harada, R., Ojima, T., Yamagami, Y., Yumoto, R. and Takano, M.
  • Journal Title: Toxicology Volume: 424 Pages: 152231-152231
  • DOI: 10.1016/j.tox.2019.06.002
  • Peer Reviewed
• [Journal Article] Reduced folate carrier-mediated methotrexate transport in human distal lung epithelial NCl-H441 cells (2019)
  • Author(s): Kawami Masashi、Honda Natsuko、Miyamoto Mioka、Yumoto Ryoko、Takano Mikihisa
  • Journal Title: Journal of Pharmacy and Pharmacology Volume: 71 Issue: 2 Pages: 167-175
  • DOI: 10.1111/jphp.13022
  • Peer Reviewed
• [Journal Article] Role of peptide transporter 2 and MAPK signaling pathways in the innate immune response induced by bacterial peptides in alveolar epithelial cells. (2019)
  • Author(s): Takano, M., Takeuchi, T., Kuriyama, S. and Yumoto, R.
  • Journal Title: Life Sci. Volume: 229 Pages: 173-179
  • DOI: 10.1016/j.lfs.2019.05.042
  • Peer Reviewed
• [Journal Article] Folic acid prevents methotrexate-induced epithelial-mesenchymal transition via suppression of secreted factors from the human alveolar epithelial cell line A549 (2018)
  • Author(s): Kawami M, Harabayashi R, Harada R, Yamagami Y, Yumoto R, Takano M
  • Journal Title: Biochem Biophys Res Commun Volume: 497 Issue: 1 Pages: 457-463
  • DOI: 10.1016/j.bbrc.2018.02.111
  • Peer Reviewed
• [Journal Article] P-gp modulating effect of Azadirachta indica extract in multidrug-resistant cancer cell lines (2018)
  • Author(s): Kawami, M., Yamada, Y., Issarachot, O., Junyaprasert, V. B., Yumoto, R., Takano, M.
  • Journal Title: Pharmazie Volume: 73 Pages: 104-109
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Transport of AOPP-albumin into human alveolar epithelial A549 cells (2018)
  • Author(s): Kawami, M., Shimonakamura, T., Yumoto, R. and Takano, M.
  • Journal Title: J. Pharm. Pharm. Sci. Volume: 21 Pages: 247-255
  • DOI: 10.18433/jpps29905
  • Peer Reviewed
• [Journal Article] Functional expression of breast cancer resistance protein and cholesterol effect in human erythrocyte membranes (2018)
  • Author(s): Takano, M., Higashi, M., Ito, H., Toyota, S., Hirabayashi, Y. and Yumoto, R.
  • Journal Title: Pharmazie Volume: 73 Pages: 700-705
  • Peer Reviewed
• [Presentation] ヒト肺胞上皮におけるmiR-34aによる上皮間葉転換誘発機構の解明 (2021)
  • Author(s): 竹中慎之介、川見昌史、湯元良子、高野幹久
  • Organizer: 日本薬学会第141年会
• [Presentation] 肺胞上皮におけるメトトレキサート誘発性上皮間葉転換とNrf2の関連解析 (2021)
  • Author(s): 本多未来一、川見昌史、湯元良子、高野幹久
  • Organizer: 日本薬学会第141年会
• [Presentation] メトトレキサート誘発性上皮間葉転換におけるITGA2の関与 (2020)
  • Author(s): 普久原梨沙、小島崇路、湯元良子、川見昌史、高野幹久
  • Organizer: 第59回日本薬学会中国四国支部学術大会
• [Presentation] Relationship between abemaciclib-induced cytotoxicity and epithelial-mesenchymal transition in a549 cells. (2020)
  • Author(s): Sorahito Futatsugi1, Ryoko Yumoto1, Masashi Kawami1, Mikihisa Takano
  • Organizer: 35nd JSSX Annual Meeting
• [Presentation] Role of high glucose and advanced glycation end products in inducing epithelial-mesenchymal-transition in alveolar epithelial cells. (2020)
  • Author(s): Izumi, Shunsuke Senoo, Masashi Kawami, Ryoko Yumoto, Mikihisa Takano
  • Organizer: 35nd JSSX Annual Meeting
• [Presentation] 高濃度グルコースおよび終末糖化産物による肺胞上皮細胞の上皮間葉転換の誘発 (2020)
  • Author(s): 泉美穂、妹尾俊祐、湯元良子、高野幹久
  • Organizer: 日本薬剤学会第36年会
• [Presentation] Role of microRNAs in drug-induced epithelial-mesenchymal transition in human lung-derived alveolar type II epithelial cell model A549/ABCA3. (2019)
  • Author(s): Ryoko Yumoto, Ayano Yamamoto, Takashi Konaka, Shinnosuke Takenaka, Masashi Kawami, Mikihisa Takano
  • Organizer: 12th International ISSX Meeting
  • Int'l Joint Research
• [Presentation] Effect of cigarette smoke extract on the functional expression of P-glycoprotein in human lung-derived A549/P-gp cells. (2019)
  • Author(s): Mikihisa Takano, Shuhei Higa, Yuma Furuichi, Ryoko Yumoto
  • Organizer: EUROTOX2019
  • Int'l Joint Research
• [Presentation] Potent inhibitory effect of vandetanib on methotrexate-induced epithelial-mesenchymal transition in alveolar epithelial A549 cells. (2019)
  • Author(s): Masashi Kawami, Ryoko Yumoto, Mikihisa Takano
  • Organizer: AAPS 2019 PharmSci 360
  • Int'l Joint Research
• [Presentation] Relationship between TGF‐β1‐induced EMT and monocarboxylate transporter 1 (MCT1) in alveolar epithelial A549 cells. (2019)
  • Author(s): Mohi Uddin, Masashi Kawami, Ryoko Yumoto, Mikihisa Takano
  • Organizer: AAPS 2019 PharmSci 360
  • Int'l Joint Research
• [Presentation] Association of methotrexate-induced epithelial-mesenchymal transition with apoptosis in human alveolar adenocarcinoma cell line A549 cells (2019)
  • Author(s): Takamichi Ojima, Ryoko Yumoto, Masashi Kawami, Mikihisa Takano
  • Organizer: 34nd JSSX Annual Meeting in Tsukuba
• [Presentation] Role of nuclear factor erythroid 2 like-2 in methotrexate-induced epithelial-mesenchymal transition in cultured human alveolar epithelial A549 cells. (2019)
  • Author(s): Takuya Hara, Ryoko Yumoto, Masashi Kawami, Mikihisa Takano
  • Organizer: 34nd JSSX Annual Meeting in Tsukuba
• [Presentation] The role of peptide transporter 2 and intracellular signaling pathways in the innate immune response induced by bacterial peptides in lung epithelial cell lines. (2019)
  • Author(s): Shiori Kuriyama, Tomoya Takeuchi, Masashi Kawami, Ryoko Yumoto, Mikihisa Takano
  • Organizer: 34nd JSSX Annual Meeting in Tsukuba
• [Presentation] Suppressive effect of vandetanib on drug-induced epithelial-mesenchymal transition via inhibition of p53 pathway (2019)
  • Author(s): Masashi Kawami, Ryoko Yumoto, Mikihisa Takano
  • Organizer: 34nd JSSX Annual Meeting in Tsukuba
• [Presentation] 肺胞上皮におけるオーファントランスポーターRhCG（SLC42A3）の特性解析 (2019)
  • Author(s): 川見昌史、渡邉光登、湯元良子、高野幹久
  • Organizer: 日本薬学会第140年会
• [Presentation] ヒト乳腺上皮細胞におけるニコチン輸送特性の解析 (2019)
  • Author(s): 渡邉光登、川見昌史、湯元良子、高野幹久
  • Organizer: 日本薬学会第140年会
• [Presentation] In vitro and In vivo study on association of miR-484 with drug-induced epithelial-mesenchymal transition. (2019)
  • Author(s): Takashi Konaka, Ayano Yamamoto, Shinnosuke Takenaka, Masashi Kawami, Ryoko Yumoto, Mikihisa Takano
  • Organizer: 第13回次世代を担う若手医療薬科学シンポジウム
• [Presentation] Role of miR-34a/p53 axis in drug-induced epithelial-mesenchymal transition in alveolar epithelial cells. (2019)
  • Author(s): Ayano Yamamoto, Masashi Kawami, Takashi Konaka, Shinnosuke Takenaka, Ryoko Yumoto, Mikihisa Takano
  • Organizer: 第13回次世代を担う若手医療薬科学シンポジウム
• [Presentation] Role of plasminogen activator inhibitor-1 in methotrexate-induced epithelial-mesenchymal transition in alveolar epithelial cells. (2019)
  • Author(s): Yohei Yamagami, Sorahito Futatsugi , Ryoko Yumoto, Masashi Kawami, Mikihisa Takano
  • Organizer: 第13回次世代を担う若手医療薬科学シンポジウム
• [Presentation] Inhibitory mechanism of cigarette smoke extract on P-glycoprotein function in human lung-derived A549/P-gp cells. (2019)
  • Author(s): Shuhei Higa, Yuma Furuichi, Ryosuke Naka, Ryoko Yumoto, Mikihisa Takano
  • Organizer: 第13回次世代を担う若手医療薬科学シンポジウム
• [Presentation] メトトレキサート誘発性上皮間葉転換に及ぼすPAI-1の影響解析 (2019)
  • Author(s): 川見昌史、山上洋平、湯元良子、高野幹久
  • Organizer: 第１１回日本RNAi研究会
• [Presentation] 肺胞上皮細胞における薬物誘発性上皮間葉転換を抑制する化合物の探索とその特性解析 (2019)
  • Author(s): 川見昌史、山上洋平、妹尾俊祐、湯元良子、高野幹久
  • Organizer: 日本薬剤学会第35年会
• [Presentation] 肺障害性薬物によるヒト肺胞上皮II型細胞モデルA549/ABCA3の上皮間葉転換とmiRNAの関連解析 (2019)
  • Author(s): 小中崇史、山本彩乃、竹中慎之介、川見昌史、湯元良子、高野幹久
  • Organizer: 日本薬剤学会第35年会
• [Presentation] 新規肺胞上皮細胞モデルを用いた肺障害性薬物による上皮間葉転換とmiR-34aの関連解析 (2019)
  • Author(s): 竹中慎之介、山本彩乃、川見昌史、湯元良子、高野幹久
  • Organizer: 第58回日本薬学会中国四国支部学術大会
• [Presentation] 肺胞上皮細胞におけるメトトレキサート誘発性上皮間葉転換に対するSrc阻害剤の影響解析 (2019)
  • Author(s): 二木空人、山上洋平、湯元良子、川見昌史、高野幹久
  • Organizer: 第58回日本薬学会中国四国支部学術大会
• [Presentation] ヒト肺胞上皮細胞モデルH441における自然免疫応答とペプチドトランスポーターPEPT2との関連解析 (2019)
  • Author(s): 栗山志織、竹内智哉、湯元良子、高野幹久
  • Organizer: 日本膜学会第41年会
• [Presentation] ニコチンの生体膜輸送におけるアンモニアトランスポーターの関与 (2019)
  • Author(s): 渡邉光登、亀井秀隆、川見昌史、湯元良子、高野幹久
  • Organizer: 日本膜学会第41年会
• [Presentation] A novel concept of nicotine transport across the cell membranes in human lung- and non-lung-derived culture cells (2018)
  • Author(s): Ryoko Yumoto, Hidetaka Kamei, Masashi Kawami and Mikihisa Takano
  • Organizer: The 8th annual Meet the Experts Transporter conference
  • Int'l Joint Research
• [Presentation] Role of plasminogen activator inhibitor-1 in methotrexate-induced epithelial-mesenchymal transition in cultured human alveolar epithelial A549 cells (2018)
  • Author(s): Yohei Yamagami、Ryoko Yumoto, Masashi Kawami, Mikihisa Takano
  • Organizer: 2018 MDO/33nd JSSX Annual Meeting in Kanazawa
  • Int'l Joint Research
• [Presentation] Role of miR-34a and p53 pathway in drug-induced epithelial-mesenchymal transition in alveolar epithelial cells (2018)
  • Author(s): Ayano Yamamoto, Masashi Kawami, Ryoko Yumoto, Mikihisa Takano
  • Organizer: 2018 MDO/33nd JSSX Annual Meeting in Kanazawa
  • Int'l Joint Research
• [Presentation] Search for the compound preventing drug-induced epithelial-mesenchymal transition and its mechanism in alveolar epithelial RLE/Abca3 cells (2018)
  • Author(s): Masashi Kawami, Takamichi Ojima, Risako Harada, Ryoko Yumoto, Mikihisa Takano
  • Organizer: 2018 MDO/33nd JSSX Annual Meeting in Kanazawa
  • Int'l Joint Research
• [Presentation] Study on association of drug-induced cell cycle arrest with epithelial-mesenchymal transition in A549 cells (2018)
  • Author(s): Shunsuke Senoo, Junya Deguchi, Ryoko Yumoto, Mikihisa Takano
  • Organizer: 2018 MDO/33nd JSSX Annual Meeting in KanazawaThe 1st Workshop of The Research Center for Drug Development and Biomarker Discovery
  • Int'l Joint Research
• [Presentation] 肺障害性薬物による肺胞上皮細胞の上皮間葉転換とmiRNAの関連解析 (2018)
  • Author(s): 川見昌史、山本彩乃、湯元良子、高野幹久
  • Organizer: 日本薬剤学会第33年会
• [Presentation] 肺胞上皮細胞におけるメトトレキサート誘発性上皮間葉転換に対する葉酸代謝機構の影響解析 (2018)
  • Author(s): 原拓也、本田奈津子、湯元良子、川見昌史、高野幹久
  • Organizer: 第57回日本薬学会中国四国支部学術大会
• [Presentation] In 肺胞上皮細胞におけるメトトレキサート誘発性上皮間葉転換に対する葉酸代謝機構の影響解析 (2018)
  • Author(s): 小島崇路、原田梨紗子、湯元良子、川見昌史、高野幹久
  • Organizer: 第57回日本薬学会中国四国支部学術大会
• [Presentation] In vitro study on secreted factors associated with drug-induced epithelial-mesenchymal transition in A549 cells (2018)
  • Author(s): Masashi Kawami, Yohei Yamagami, Risako Harada, Takamichi Ojima, Ryoko Yumoto, Mikihisa Takano
  • Organizer: The 1st Workshop of The Research Center for Drug Development and Biomarker Discovery
  • Int'l Joint Research
• [Presentation] 肺胞上皮細胞に対するメトトレキサートの抗がん効果と上皮間葉転換誘発作用の関連解析 (2018)
  • Author(s): 川見昌史、原田梨紗子、小島崇路、湯元良子、高野幹久
  • Organizer: 日本薬学会第139年会