A third generation of genetically encoded fluorescent lactate biosensor

• Project/Area Number: 21K14738
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: The 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): 2021-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
• Keywords: 乳酸 / 蛍光タンパク質 / 蛍光バイオセンサー / Metabolism

Outline of Research at the Start

本研究では, 「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の常識を覆す説を検証可能にする第三世代蛍光乳酸センサーの開発を行う.

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 third-generation genetically encoded fluorescent biosensor for extracellular L-lactate by using 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, 高時空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。

Research Products

• [Int'l Joint Research] Janelia Research Campus/Allen Institute/Montana State University(米国)
• [Int'l Joint Research] Laval University/University of Alberta(カナダ)
• [Int'l Joint Research] INSERM(フランス)
• [Int'l Joint Research] カルガリー大学/モントリオール大学/アルバータ大学(カナダ)
• [Int'l Joint Research] モンタナ州立大学/ハワードヒューズ医学研究所Janelia Research Campus(米国)
• [Int'l Joint Research] 西安交通大学(中国)
• [Journal Article] The endoplasmic reticulum kinase PERK interacts with the oxidoreductase ERO1 to metabolically adapt mitochondria (2023)
  • Author(s): Bassot Arthur、Chen Junsheng、Takahashi-Yamashiro Kei、他17名
  • Journal Title: Cell Reports Volume: 42 Issue: 1 Pages: 111899-111899
  • DOI: 10.1016/j.celrep.2022.111899
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A lactate-dependent shift of glycolysis mediates synaptic and cognitive processes (2023)
  • Author(s): Ignacio Fernandez-Moncada et al.
  • Journal Title: bioRxiv Volume: -
  • DOI: 10.1101/2023.03.15.532748
  • Open Access / Int'l Joint Research
• [Journal Article] Chemigenetic indicators based on synthetic chelators and green fluorescent protein (2022)
  • Author(s): Zhu Wenchao、Takeuchi Shiori、Imai Shosei、Terada Tohru、Ueda Takumi、Nasu Yusuke、Terai Takuya、Campbell Robert E.
  • Journal Title: Nature Chemical Biology Volume: 19 Issue: 1 Pages: 38-44
  • DOI: 10.1038/s41589-022-01134-z
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A genetically encoded far‐red fluorescent calcium ion biosensor derived from a biliverdin‐binding protein (2022)
  • Author(s): Hashizume Rina、Fujii Hajime、Mehta Sohum、Ota Keisuke、Qian Yong、Zhu Wenchao、Drobizhev Mikhail、Nasu Yusuke、Zhang Jin、Bito Haruhiko、Campbell Robert E.
  • Journal Title: Protein Science Volume: 31 Issue: 10
  • DOI: 10.1002/pro.4440
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Improved genetically encoded fluorescent biosensors for monitoring of intra- and extracellular L-lactate (2022)
  • Author(s): Nasu Y.*, Aggarwal A., Le G. N. T., Kamijo Y., Boisvert M., Paquet M-E., Drobizhev M., Podgorski K., Campbell R. E.*
  • Journal Title: bioRxiv Volume: -
  • DOI: 10.1101/2022.12.27.522013
  • Open Access / Int'l Joint Research
• [Journal Article] High performance genetically-encoded green fluorescent biosensors for intracellular L-lactate (2022)
  • Author(s): Le G. N. T., Hario S., Takahashi-Yamashiro K., Li S., Drobizhev M., Nasu Y.*, Campbell R. E.*
  • Journal Title: bioRxiv Volume: -
  • DOI: 10.1101/2022.10.19.512892
  • Open Access / Int'l Joint Research
• [Journal Article] A red fluorescent genetically encoded biosensor for extracellular L-lactate (2022)
  • Author(s): Nasu Yusuke、Kamijo Yuki、Hashizume Rina、Sato Haruaki、Hori Yukiko、Tomita Taisuke、Drobizhev Mikhail、Campbell Robert E.
  • Journal Title: bioRxiv Volume: -
  • DOI: 10.1101/2022.08.30.505811
  • Open Access / Int'l Joint Research
• [Journal Article] A genetically encoded fluorescent biosensor for extracellular L-lactate (2021)
  • Author(s): Yusuke Nasu, Ciaran Murphy-Royal, Yurong Wen, Jordan N. Haidey, Rosana S. Molina, Abhi Aggarwal, Shuce Zhang, Yuki Kamijo, Marie-Eve Paquet, Kaspar Podgorski, Mikhail Drobizhev, Jaideep S. Bains, M. Joanne Lemieux, Grant R. Gordon & Robert E. Campbell
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 7058-7058
  • DOI: 10.1038/s41467-021-27332-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 高感度蛍光乳酸バイオセンサー -LACCOシリーズ- (2023)
  • Author(s): Nasu Y., Kamijo Y., Le G., Hario S., and Robert E. Campbell
  • Organizer: 日本化学会 第102春季年会
• [Presentation] LACCO series: Genetically encoded fluorescent biosensors for extracellular and intracellular L-lactate (2022)
  • Author(s): Nasu Y., Kamijo Y., Hario S., Le G. N. T., and Campbell R. E.
  • Organizer: Society for Neuroscience 2022
  • Int'l Joint Research
• [Presentation] Lactate matters: Genetically encoded lactate biosensors (2022)
  • Author(s): Nasu Y.
  • Organizer: Serendipity Workshop 2022
  • Invited
• [Presentation] Genetically encoded fluorescent biosensors for neuroscience (2022)
  • Author(s): Nasu Y.
  • Organizer: Multiphoton Workshop 2022
  • Int'l Joint Research
• [Presentation] 乳酸バイオセンサーの開発 (2022)
  • Author(s): Nasu Y., Le G., Hario S., Kamijo Y., and Robert E. Campbell
  • Organizer: 第16回バイオ関連化学シンポジウム
• [Presentation] High-performance genetically encoded biosensors for L-lactate (2022)
  • Author(s): Nasu Y., Le G., Hario S., Kamijo Y., and Robert E. Campbell
  • Organizer: The 45th Annual Meeting of the Molecular Biology Society of Japan
  • Invited
• [Presentation] 乳酸バイオセンサーの開発 (2022)
  • Author(s): 那須雄介, 針尾紗彩, Robert E. Campbell
  • Organizer: 日本化学会 第102春季年会
• [Presentation] An ultrasensitive fluorescent biosensor for L-lactate (2021)
  • Author(s): Nasu Y., and Robert E. Campbell
  • Organizer: Pacifichem
  • Int'l Joint Research
• [Presentation] Engineering of genetically encoded fluorescent sensors (2021)
  • Author(s): Nasu Y., and Robert E. Campbell
  • Organizer: Pacifichem
  • Int'l Joint Research
• [Book] 月刊細胞 (2023)
  • Author(s): 那須雄介
  • Total Pages: 4
  • Publisher: ニューサイエンス社
• [Book] 化学 (2022)
  • Author(s): 那須 雄介
  • Total Pages: 2
  • Publisher: 化学同人
• [Book] 先端の分析法 新訂第２版 (2022)
  • Author(s): 那須 雄介
  • Total Pages: 8
  • Publisher: NTS
  • ISBN: 9784860437374
• [Remarks] ORCID
  • URL: https://orcid.org/0000-0002-3193-9104
• [Remarks] Researchmap
  • URL: https://researchmap.jp/yusukenasu
• [Remarks] 東大、細胞外の乳酸を緑色蛍光で低侵襲に可視化するバイオセンサーeLACCO1.1を開発
  • URL: https://www.nikkei.com/article/DGXLRSP622938_S1A201C2000000/
• [Remarks] 細胞外乳酸バイオセンサーの開発
  • URL: https://www.s.u-tokyo.ac.jp/ja/press/2021/7657/