Drug screening using a newly developed cell assay

• Project/Area Number: 16K12872
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biomedical engineering/Biomaterial science and engineering
• Research Institution: Osaka University
• Principal Investigator: Deguchi Shinji 大阪大学, 基礎工学研究科, 教授 (30379713)
• Research Collaborator: Matsui Tsubasa S.
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 細胞バイオメカニクス / 細胞収縮力 / メカノミクス / ドラッグリポジショニング / ドラッグ・リポジショニング / 創薬 / メカノーム

Outline of Final Research Achievements

Cells regulate their function according to the contractile force that the individual cells generate. However, it remains unclear regarding the detailed relationship between the cell functions and contractile force because of the lack of methodologies allowing comprehensive analysis on the relationship. Here, we have developed a new method that allows us to quantitatively evaluate the cellular contractile force with a high-throughput analysis capability. We demonstrated with the new system that some molecular regulators are found to be effective for modulating the cellular contractile force.

Academic Significance and Societal Importance of the Research Achievements

細胞の「収縮」に関する研究と言えばこれまで主に筋肉（骨格筋、心筋、平滑筋）に関するものが主なものでした。しかし昨今、「非筋」細胞における微小な収縮が、広範な細胞機能の調節に重要な役割を果たしていることが明らかにされています。本研究では非筋細胞の収縮の程度を高い実験効率で評価できる新しい装置の開発を行いました。この装置を用いた実験により、非筋細胞の収縮を制御する薬剤の同定を行いました。これにより非筋細胞内の収縮能の調節およびそれに依存した諸機能の分子機構の解明につながることが期待されます。

Research Products

• [Journal Article] Keratin-binding ability of the N-terminal Solo domain of Solo is critical for its function in cellular mechanotransduction (2019)
  • Author(s): Fujiwara, S., Matsui, T. S., Ohashi, K., Mizuno, K., and Deguchi, S.
  • Journal Title: Genes Cells Volume: 印刷中 Issue: 5 Pages: 390-402
  • DOI: 10.1111/gtc.12682
  • Peer Reviewed
• [Journal Article] Spatially selective MRLC regulation is absent in dedifferentiated vascular smooth muscle cells but is partially induced by fibronectin and Klf4 (2019)
  • Author(s): Matsui, T.S., Deguchi, S.
  • Journal Title: American Journal of Physiology Cell Physiology Volume: 316 Issue: 4 Pages: C509-C521
  • DOI: 10.1152/ajpcell.00251.2017
  • Peer Reviewed
• [Journal Article] Transgelin-1 (SM22α) interacts with actin stress fibers and podosomes in smooth muscle cells without using its actin binding site (2018)
  • Author(s): Matsui, T.S., Ishikawa, A., Deguchi, S.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 505 Issue: 3 Pages: 879-884
  • DOI: 10.1016/j.bbrc.2018.09.176
  • Peer Reviewed
• [Journal Article] Deformable 96-well cell culture plate compatible with high-throughput screening platforms (2018)
  • Author(s): Matsui, T.S., Wu, H., Deguchi, S.
  • Journal Title: PLOS ONE Volume: 13 Issue: 9 Pages: e0203448-e0203448
  • DOI: 10.1371/journal.pone.0203448
  • Peer Reviewed / Open Access
• [Journal Article] Solo, a RhoA-targeting guanine nucleotide exchange factor, is critical for hemidesmosome formation and acinar development in epithelial cells (2018)
  • Author(s): Sachiko Fujiwara, Tsubasa S. Matsui, Kazumasa Ohashi, Shinji Deguchi, and Kensaku Mizuno
  • Journal Title: PLOS ONE Volume: 印刷中 Issue: 4 Pages: e0195124-e0195124
  • DOI: 10.1371/journal.pone.0195124
  • Peer Reviewed / Open Access
• [Journal Article] Vinexin family (SORBS) proteins play different roles in stiffness-sensing and contractile force generation (2017)
  • Author(s): Ichikawa, T., Kita, M., Matsui, T.S., Ichikawa-Nagasato, A., Araki, T., Chiang, S.H., Sezaki, T., Kimura, Y., Ueda, K., Deguchi, S., Saltiel, A.R., Kioka, N.
  • Journal Title: Journal of Cell Science Volume: 130 Issue: 20 Pages: 3517-3531
  • DOI: 10.1242/jcs.200691
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Cellular force assay detects altered contractility caused by a nephritis-associated mutation in nonmuscle myosin IIA (2017)
  • Author(s): Fukuda, S.P., Matsui, T.S., Ichikawa, T., Furukawa, T., Kioka, N., Fukushima, S., Deguchi, S.
  • Journal Title: Development, Growth & Differentiation Volume: 59 Issue: 5 Pages: 423-433
  • DOI: 10.1111/dgd.12379
  • Peer Reviewed
• [Journal Article] New wrinkling substrate assay reveals traction force fields of leader and follower cells undergoing collective migration (2017)
  • Author(s): Yokoyama, S., Matsui, T.S., Deguchi, S.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 482 Issue: 4 Pages: 975-979
  • DOI: 10.1016/j.bbrc.2016.11.142
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Microcontact Peeling: A Cell Micropatterning Technique for Circumventing Direct Adsorption of Proteins to Hydrophobic PDMS (2017)
  • Author(s): Yokoyama, S., Matsui, T.S., Deguchi, S.
  • Journal Title: Current Protocols in Cell Biology Volume: 75
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Tensile strength of oxygen plasma-created surface layer of PDMS (2017)
  • Author(s): Ohishi, T., Noda, H., Matsui, T.S., Jile, H., Deguchi, S.
  • Journal Title: Journal of Micromechanics and Microengineering Volume: 27 Pages: 015015-015015
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Conformational Plasticity of JRAB/MICAL-L2 Provides 'Law and Order' (2016)
  • Author(s): A. Sakane, S. Yoshizawa, M. Nishimura, Y. Tsuchiya, N. Matsushita, K. Miyake, K. Horikawa, I. Imoto, C. Mizuguchi, H. Saito, T. Ueno, S. Matsushita, H. Haga, S. Deguchi, K. Mizuguchi, H.
  • Journal Title: Molecular Biology of the Cell Volume: 27 Issue: 20 Pages: 3095-3108
  • DOI: 10.1091/mbc.e16-05-0332
  • Peer Reviewed / Open Access
• [Presentation] A cellular contractility monitoring system allowing for high- throughput and long-term analysis (2018)
  • Author(s): T.S. Matsui, S. Deguchi
  • Organizer: MHS 2018 (International Symposium on Micro-NanoMechatronics and Human Science)
  • Int'l Joint Research
• [Presentation] Rho-signaling organizes hemidesmosomal cell-substrate adhesion and acinar morphogenesis of epithelial cells through controlling mechanotransduction (2018)
  • Author(s): S. Fujiwara, T.S. Matsui, K. Ohashi, T.M. Magin, K. Mizuno, S. Deguchi
  • Organizer: International Meeting of the German Society for Cell Biology
  • Int'l Joint Research
• [Presentation] High-throughput detection of cellular traction forces for screening of drugs and regulatory genes (2018)
  • Author(s): Deguchi, S.
  • Organizer: Emerging Technologies in Mechanical Engineering 2018
  • Int'l Joint Research / Invited
• [Presentation] Stress fibers exhibit unique contractile properties distinct from those of myofibrils (2018)
  • Author(s): Deguchi, S., Matsui, T.S
  • Organizer: 8th World Congress of Biomechanics
  • Int'l Joint Research
• [Presentation] Hemidesmosomal cell-substrate adhesion and acinar morphogenesis of epithelial cells are organized by Rho-signaling through controlling mechanotransduction (2018)
  • Author(s): S. Fujiwara, T. S. Matsui, K. Ohashi, K. Mizuno, S. Deguchi
  • Organizer: 8th World Congress of Biomechanics
  • Int'l Joint Research
• [Presentation] Rho-signaling plays crucial roles for cellular mechanotransduction and regulates actin and intermediate filament cytoskeletal reorganization (2018)
  • Author(s): S. Fujiwara, H. Abiko, T. S. Matsui, K. Ohashi, K. Mizuno, S. Deguchi
  • Organizer: 8th World Congress of Biomechanics
  • Int'l Joint Research
• [Presentation] Isometric contractile properties of individual stress fibers (2018)
  • Author(s): Deguchi, S.
  • Organizer: 8th World Congress of Biomechanics
  • Int'l Joint Research / Invited
• [Presentation] スループットの高い細胞トラクションフォースアッセイの開発 (2017)
  • Author(s): 出口 真次，松井 翼，田中 裕一郎，呼格吉楽
  • Organizer: 第29回バイオエンジニアリング講演会
  • Place of Presentation: ウインクあいち
  • Year and Date: 2017-01-19
• [Presentation] Detecting endogenous forces of cells subjected to mutations and drugs (2017)
  • Author(s): S. Deguchi
  • Organizer: 第76回 日本癌学会学術総会
  • Int'l Joint Research / Invited
• [Presentation] Local geometry sensing by individual focal adhesions (2017)
  • Author(s): Shinji Deguchi, Sho Yokoyama, Tsubasa S. Matsui, Taiki Ohishi
  • Organizer: 2017 Cellular and Molecular Bioengineering Annual Conference
  • Int'l Joint Research
• [Presentation] 個々の細胞が発生する力を高効率に定量化するシステムの開発 (2016)
  • Author(s): 松井 翼, 出口 真次
  • Organizer: 第39回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2016-11-30
• [Presentation] ミオシン調節軽鎖のリン酸化は個々の細胞が発生する収縮力の大きさに相関はあるか？ (2016)
  • Author(s): 出口 真次, 松井 翼, 倉賀野 正弘, 高橋 正行
  • Organizer: 第68回日本細胞生物学会大会
  • Place of Presentation: 京都テルサ
  • Year and Date: 2016-06-15
• [Book] 細胞収縮力アッセイ in 月刊「細胞」 (2019)
  • Author(s): 出口 真次，松井 翼，市川 尚文，木岡 紀幸
  • Total Pages: 4
  • Publisher: 北隆館 ニューサイエンス社
• [Book] 細胞収縮力のアッセイ － メカノバイオロジー研究のツール in Precision Medicine プレシジョンメディシンと創薬 (2018)
  • Author(s): 出口 真次，松井 翼，市川 尚文，木岡 紀幸
  • Total Pages: 4
  • Publisher: 北隆館 ニューサイエンス社
• [Remarks] 大阪大学・出口研
  • URL: http://mbm.me.es.osaka-u.ac.jp
• [Remarks] 大阪大学大学院基礎工学研究科・出口研HP
  • URL: http://mbm.me.es.osaka-u.ac.jp
• [Remarks] 大阪大学・出口研究室ホームページ
  • URL: http://mbm.me.es.osaka-u.ac.jp