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Development of Ultrasensitive Fluorescent Lactate Indicator for Intercellular Metabolism

Research Project

Project/Area Number 19K15691
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 37010:Bio-related chemistry
Research InstitutionThe University of Tokyo

Principal Investigator

Nasu Yusuke  東京大学, 大学院理学系研究科(理学部), 助教 (60831328)

Project Period (FY) 2019-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2019: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Keywords蛍光タンパク質 / 乳酸 / 蛍光バイオセンサー / タンパク質工学 / 蛍光センサー / Metabolism
Outline of Research at the Start

「生物の主要な細胞間エネルギー伝達物質はグルコースではなく乳酸である」というこれまでのmetabolismの常識を覆す説が近年提唱されている. この「非常識」を検証するには細胞間の乳酸動態を観察する必要があり, そのためには「細胞内」及び「細胞外」の乳酸を観察可能なセンサーが必要である. これまで細胞内外共に生体観察可能な乳酸センサーは全くなく, その開発が急務であった. これまでの研究で, 申請者はタンパク質工学の手法を駆使して「細胞内」乳酸蛍光センサーG-iLACCO1を開発した. そこで本研究では, 「細胞外」乳酸蛍光センサーの開発を目的とする.

Outline of Final Research Achievements

L-Lactate, traditionally considered a metabolic waste product, is increasingly recognized as an important intercellular energy currency in mammals. Investigations of the emerging roles of intercellular shuttling of L-lactate requires a method to observe extracellular L-lactate in living tissue with high spatiotemporal resolution. To achieve this goal, we developed a genetically encoded red fluorescent biosensor for extracellular L-lactate by using technique of protein engineering.

Academic Significance and Societal Importance of the Research Achievements

これまで細胞外の乳酸動態を観察する方法は, 血液を採取して生化学的に解析したり, 観察部位に乳酸反応性電極を挿入したりなど, 乳酸に関する時空間情報の欠如や生体に対する侵襲性が避けられなかった。世界で初めて細胞外乳酸を非侵襲的に, かつ, 高時間分解能で観察することが可能なバイオセンサーを開発した, という点で本研究の成果は学術的に極めて意義がある。さらに, 乳酸は社会的な認知度・注目度も高いことから, 本研究成果である細胞外乳酸センサーを利用することでこれまで知られていなかった乳酸の新たな知見の発見につながる可能性がある点で社会的な意義がある。

Report

(3 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • Research Products

    (6 results)

All 2021 2020 2019

All Journal Article (3 results) (of which Int'l Joint Research: 3 results,  Open Access: 1 results,  Peer Reviewed: 2 results) Presentation (3 results) (of which Int'l Joint Research: 3 results)

  • [Journal Article] A genetically encoded fluorescent biosensor for extracellular L-lactate2021

    • Author(s)
      Y. Nasu, C. Murphy-Royal, Y. Wen, J. Haidey, M.R.S. Molina, A. Aggarwal, S. Zhang, Y. Kamijo, M.-E. Paquet, K. Podgorski, M. Drobizhev, J.S. Bains, M.J. Lemieux, G.R. Gordon, R.E. Campbell
    • Journal Title

      bioRxiv

      Volume: N/A

    • DOI

      10.1101/2021.03.05.434048

    • Related Report
      2020 Annual Research Report
    • Open Access / Int'l Joint Research
  • [Journal Article] Structure- and mechanism-guided design of single fluorescent protein-based biosensors2021

    • Author(s)
      Nasu Y., Shen Y., Kramer L., Campbell R.E.
    • Journal Title

      Nature Chemical Biology

      Volume: N/A Issue: 5 Pages: 509-518

    • DOI

      10.1038/s41589-020-00718-x

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Engineering genetically encoded fluorescent indicators for imaging of neuronal activity: Progress and prospects2020

    • Author(s)
      Shen Yi、Nasu Yusuke、Shkolnikov Irene、Kim Anna、Campbell Robert E.
    • Journal Title

      Neuroscience Research

      Volume: 152 Pages: 3-14

    • DOI

      10.1016/j.neures.2020.01.011

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] An ultrasensitive fluorescent biosensor for L-lactate2021

    • Author(s)
      Nasu Y. and Campbell R.E.
    • Organizer
      Pacifichem
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Development of Genetically Encoded Fluorescent Indicator2020

    • Author(s)
      Nasu Y., and Robert E. Campbell
    • Organizer
      The 4th ETH Zurich UTokyo Strategic Partnership Symposium
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research
  • [Presentation] Development of a genetically encoded intensiometric lactate indicator iLACCO12019

    • Author(s)
      Nasu Y., and Robert E. Campbell
    • Organizer
      ACS Fall 2019 National Meeting & Exposition
    • Related Report
      2019 Research-status Report
    • Int'l Joint Research

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Published: 2019-04-18   Modified: 2022-12-28  

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