Development of microbiosensor with DNA scaffolds for cell metabolic analysis

• Project/Area Number: 17K05917
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Analytical chemistry
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: Hirano Yu 国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (70415735)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 電気化学 / 細胞 / 酵素 / マイクロ電極 / DNA / SECM / 心筋細胞 / マイクロ・ナノ電極

Outline of Final Research Achievements

Metabolic pathways are a series of chemical reactions that occur within a cell. Since cellular metabolism is an indicator to analyze living cells, evaluation of the effect on metabolic activity is essential for drug development. In this study, we developed a system to analyze the concentration of metabolites in the vicinity of cells by immobilizing enzymes on a tip of a needle-shaped microelectrode. In combination with scanning electrochemical microscopy (SECM), it was possible to evaluate the glucose consumption of living cells.

Academic Significance and Societal Importance of the Research Achievements

培養した細胞を利用すると薬剤の効果などを評価することができるが、細胞単位で細胞が消費や生産したりする生体分子を定量することは難しかった。本研究では、細胞の薬剤添加に伴う変化を連続して評価できる顕微鏡システムを開発した。特に、細胞サイズの小さな電極に酵素などを固定化することで、細胞が消費するグルコースなどの定量が可能となった。開発したシステムを利用して、心筋細胞などに対する薬剤効果の評価に応用した。本研究の成果は、将来的には医薬品開発や基礎研究に役立つことが期待できる。

Research Products

• [Journal Article] Facile control of surface crystallographic orientation of anodized nanoporous gold catalyst and its application for highly efficient hydrogen evolution reaction (2020)
  • Author(s): Mie Yasuhiro、Takayama Haruka、Hirano Yu
  • Journal Title: Journal of Catalysis Volume: 389 Pages: 476-482
  • DOI: 10.1016/j.jcat.2020.06.023
  • Peer Reviewed
• [Journal Article] Inhibition of breast cancer cell proliferation with anti-microRNA oligonucleotides flanked by interstrand cross-linked duplexes (2020)
  • Author(s): Okumura Sho、Hirano Yu、Komatsu Yasuo
  • Journal Title: Nucleosides, Nucleotides & Nucleic Acids Volume: 39 Issue: 1-3 Pages: 225-235
  • DOI: 10.1080/15257770.2019.1671595
  • Peer Reviewed
• [Journal Article] Adsorptive Stripping Voltammetry for the Determination of Dissolved Oxygen Using a Mesoporous Pt Microelectrode (2019)
  • Author(s): Masiki Ikegami, Yu Hirano, Yasuhiro Mie, Yasuo Komatsu
  • Journal Title: Journal of The Electrochemical Society Volume: 166 Issue: 6 Pages: B542-B546
  • DOI: 10.1149/2.0021908jes
  • Peer Reviewed
• [Journal Article] Analysis of time-course drug response in rat cardiomyocytes cultured on a pattern of islands (2018)
  • Author(s): Okumura Sho、Hirano Yu、Maki Yoshiyuki、Komatsu Yasuo
  • Journal Title: The Analyst Volume: 143 Issue: 17 Pages: 4083-4089
  • DOI: 10.1039/c8an01033a
  • NAID: 120006718705
  • Peer Reviewed
• [Journal Article] Synthesis and Application of Interstrand Cross-Linked Duplexes by Covalently Linking a Pair of Abasic Sites (2018)
  • Author(s): Hirano Yu、Kojima Naoshi、Komatsu Yasuo
  • Journal Title: Current Protocols in Nucleic Acid Chemistry Volume: 75 Issue: 1
  • DOI: 10.1002/cpnc.63
  • Peer Reviewed
• [Journal Article] Function Control of Anti-microRNA Oligonucleotides Using Interstrand Cross-Linked Duplexes (2018)
  • Author(s): Yasuhiro Mie, Yu Hirano, Keiko Kowata, Akiyoshi Nakamura, Mayu Yasunaga, Yoshihiro Nakajima, Yasuo Komatsu
  • Journal Title: MOLECULAR THERAPY-NUCLEIC ACIDS Volume: 10 Pages: 64-74
  • DOI: 10.1016/j.omtn.2017.11.003
  • Peer Reviewed / Open Access
• [Presentation] クロスリンク２本鎖構造によるanti-miRNAオリゴの細胞内局在への影響 (2019)
  • Author(s): 平野 悠、扇谷 仁希、千高 佐知子、三重 安弘、小松 康雄
  • Organizer: 核酸医薬学会 第4回年会
• [Presentation] Effect of terminal double-stranded structures on intracellular localization of anti-miRNA oligonucleotide (2019)
  • Author(s): 平野 悠、千高 佐知子、小松 康雄
  • Organizer: International Symposium on Nucleic Acids Chemistry 2019
  • Int'l Joint Research
• [Presentation] クロスリンク２本鎖構造を有するanti-miRNAオリゴ核酸の細胞内動態 (2018)
  • Author(s): 平野 悠, 扇谷仁希, 三重安弘, 小綿 恵子, 小松 康雄
  • Organizer: 核酸医薬学会 第4回年会
• [Presentation] 微細構造白金マイクロ電極を用いた溶存酸素の検出 (2018)
  • Author(s): 池上 真志樹, 平野 悠, 三重 安弘
  • Organizer: 応用物理学会
• [Presentation] Construction of a bienzyme immobilizing microelectrode by cross-linked DNA scaffolds and its application to analysis of cellular metabolism (2017)
  • Author(s): 平野 悠, 池上 真志樹, 小綿 恵子, 小松 康雄
  • Organizer: The 44th International Symposium on Nucleic Acids Chemistry 2017
  • Int'l Joint Research
• [Presentation] 走査型電気化学顕微鏡用の白金/金コンポジット微細構造電極の開発 (2017)
  • Author(s): 池上 真志樹, 平野 悠, 三重 安弘, 小松 康雄
  • Organizer: 第65回応用物理学会春季学術講演会
• [Book] 不凍タンパク質の機能と応用 (2018)
  • Author(s): 平野悠 ほか （監修 津田栄）
  • Total Pages: 290
  • Publisher: シーエムシー出版
  • ISBN: 9784781313399