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Development of functional fluorescent probe for imaging osteocyte function and 4D nucleomics

Research Project

Project/Area Number 21H04706
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Review Section Medium-sized Section 37:Biomolecular chemistry and related fields
Research InstitutionOsaka University

Principal Investigator

Kazuya Kikuchi  大阪大学, 大学院工学研究科, 教授 (70292951)

Project Period (FY) 2021-04-05 – 2024-03-31
Project Status Completed (Fiscal Year 2023)
Budget Amount *help
¥42,120,000 (Direct Cost: ¥32,400,000、Indirect Cost: ¥9,720,000)
Fiscal Year 2023: ¥13,390,000 (Direct Cost: ¥10,300,000、Indirect Cost: ¥3,090,000)
Fiscal Year 2022: ¥13,390,000 (Direct Cost: ¥10,300,000、Indirect Cost: ¥3,090,000)
Fiscal Year 2021: ¥15,340,000 (Direct Cost: ¥11,800,000、Indirect Cost: ¥3,540,000)
Keywords化学プローブ / 蛍光イメージング / 蛋白質ラベル化 / 破骨細胞 / 蛍光プローブ / in vivoイメージング / pH計測 / 蛋白質局在 / タンパク質ラベル化 / 骨細胞 / オルガネラ成熟 / ゲノム動態可視化
Outline of Research at the Start

本研究では、細胞内分子機能解明に加え生物個体における細胞機能を観察できる化学ツールの設計法を確立する。これまでの蛍光プローブ開発の発展型として化学分子設計と蛋白質科学技術と融合することで、in vivo解析へ応用可能な化学プローブへと深化させる。最終的にこれらのプローブを細胞から動物個体へ応用し、生物学における問題解決に応用する。

Outline of Final Research Achievements

In this study, we developed functional fluorescent probes to directly measure physiological functions of living cells. The aim of this project is to reveal biological phenomena that can be functionally analyzed for the first time with this technology by combining organic chemistry and cell biology. During this project, we have developed in vivo imaging probes that reveal osteocyte/osteoclast functions using two-photon excitation microscopy. Also, we have developed novel fluorescent protein labeling probes for visualizing genome and organelle dynamics. Our results have enabled applications to the analysis of cellular functions in living animals or protein dynamics in living cells.

Academic Significance and Societal Importance of the Research Achievements

本研究は細胞や生体内の分子機能を生きた状態で観察するための化学プローブの開発を行うことで、初めて観察できる現象を捉えることを目的としている。そのため、化学を基にしたpHに応答し波長が変化する蛍光プローブ、標的蛋白質を速やかに長時間観察できるラベル化プローブを開発することに成功した。実際に、これらのプローブを用いて生きたマウス骨組織におけるpHの変化や、細胞内における蛋白質の移行過程を可視化することに成功しており、生物学、医学分野への貢献が期待できる成果が得られた。

Report

(5 results)
  • 2023 Annual Research Report   Final Research Report ( PDF )
  • 2022 Annual Research Report
  • 2021 Comments on the Screening Results   Annual Research Report
  • Research Products

    (32 results)

All 2024 2023 2022 2021

All Journal Article (22 results) (of which Int'l Joint Research: 9 results,  Peer Reviewed: 21 results,  Open Access: 12 results) Presentation (6 results) (of which Int'l Joint Research: 5 results,  Invited: 6 results) Book (1 results) Patent(Industrial Property Rights) (3 results)

  • [Journal Article] Activatable 19F MRI probe for detection of α-amylase activity2024

    • Author(s)
      Konishi Yuki、Otera Yuya、Minoshima Masafumi、Kikuchi Kazuya
    • Journal Title

      Chemistry Letters

      Volume: 53 Issue: 1

    • DOI

      10.1093/chemle/upae003

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rational Design of Hydroxylated Thiazole Orange Photocages for Green Light‐Triggered DNA Recombination2024

    • Author(s)
      Hashimoto Ryu、Minoshima Masafumi、Kikuchi Kazuya
    • Journal Title

      ChemBioChem

      Volume: 25 Issue: 4

    • DOI

      10.1002/cbic.202300799

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Recent advancements of fluorescent biosensors using semisynthetic probes2024

    • Author(s)
      Reja Shahi Imam、Minoshima Masafumi、Hori Yuichiro、Kikuchi Kazuya
    • Journal Title

      Biosensors and Bioelectronics

      Volume: 247 Pages: 115862-115862

    • DOI

      10.1016/j.bios.2023.115862

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] No-wash fluorogenic labeling of proteins for reversible photoswitching in live cells2024

    • Author(s)
      Torii Kenji、Benson Sam、Hori Yuichiro、Vendrell Marc、Kikuchi Kazuya
    • Journal Title

      Chemical Science

      Volume: 15 Issue: 4 Pages: 1393-1401

    • DOI

      10.1039/d3sc04953a

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Elastic Polymer Coated Nanoparticles with Fast Clearance for 19F MR Imaging2023

    • Author(s)
      Konishi Yuki、Minoshima Masafumi、Fujihara Kohei、Uchihashi Takayuki、Kikuchi Kazuya
    • Journal Title

      Angewandte Chemie International Edition

      Volume: 62 Issue: 40

    • DOI

      10.1002/anie.202308565

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Development of a Versatile Protein Labeling Tool for Live‐Cell Imaging Using Fluorescent β‐Lactamase Inhibitors2023

    • Author(s)
      Minoshima Masafumi、Umeno Taro、Kadooka Kohei、Roux Margaux、Yamada Namiko、Kikuchi Kazuya
    • Journal Title

      Angewandte Chemie International Edition

      Volume: 62 Issue: 18

    • DOI

      10.1002/anie.202301704

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Visualization of multiple localizations of GLUT4 by fluorescent probes of PYP-tag with designed unnatural warhead2023

    • Author(s)
      Nishiura Miyako、Hori Yuichiro、Umeno Maho、Kikuchi Kazuya
    • Journal Title

      Chemical Science

      Volume: 14 Issue: 22 Pages: 5925-5935

    • DOI

      10.1039/d3sc00724c

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Persistent Fluorescence Switching of a Probe Using a Photochromic Quencher with High Photostability Assisted by Protein-Surface Modification2023

    • Author(s)
      Torii Kenji、Hori Yuichiro、Kikuchi Kazuya
    • Journal Title

      Analytical Chemistry

      Volume: 95 Issue: 23 Pages: 8834-8841

    • DOI

      10.1021/acs.analchem.3c00163

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Long-term imaging of intranuclear Mg<sup>2+</sup> dynamics during mitosis using a localized fluorescent probe2023

    • Author(s)
      Matsui Yusuke、Kowada Toshiyuki、Ding Yi、Sahoo Priya Ranjan、Kikuchi Kazuya、Mizukami Shin
    • Journal Title

      Chemical Communications

      Volume: 59 Issue: 46 Pages: 7048-7051

    • DOI

      10.1039/d2cc05930d

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Variational autoencoder-based chemical latent space for large molecular structures with 3D complexity2023

    • Author(s)
      Ochiai Toshiki、Inukai Tensei、Akiyama Manato、Furui Kairi、Ohue Masahito、Matsumori Nobuaki、Inuki Shinsuke、Uesugi Motonari、Sunazuka Toshiaki、Kikuchi Kazuya、Kakeya Hideaki、Sakakibara Yasubumi
    • Journal Title

      Communications Chemistry

      Volume: 6 Issue: 1 Pages: 249-249

    • DOI

      10.1038/s42004-023-01054-6

    • Related Report
      2023 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] 生命機能を可視化するさまざまな化学蛍光プローブ2023

    • Author(s)
      蓑島維文、菊地和也
    • Journal Title

      医学のあゆみ 生体イメージングの最前線

      Volume: 286 Pages: 337-342

    • Related Report
      2023 Annual Research Report
  • [Journal Article] An “OFF-ON-OFF” fluorescence protein-labeling probe for real-time visualization of the degradation of short-lived proteins in cellular systems2022

    • Author(s)
      Reja Shahi Imam、Hori Yuichiro、Kamikawa Takuya、Yamasaki Kohei、Nishiura Miyako、Bull Steven D.、Kikuchi Kazuya
    • Journal Title

      Chemical Science

      Volume: 13 Issue: 5 Pages: 1419-1427

    • DOI

      10.1039/d1sc06274c

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] pH‐Sensitive Polymethacrylates as Potential Contrast Agents in 19F MRI2022

    • Author(s)
      Zalewski Mariusz、Janasik Dawid、Kapala Anna、Minoshima Masafumi、Sugihara Fuminori、Raj Wojciech、Pietrasik Joanna、Kikuchi Kazuya、Krawczyk Tomasz
    • Journal Title

      Macromolecular Chemistry and Physics

      Volume: 223 Issue: 14 Pages: 2200027-2200027

    • DOI

      10.1002/macp.202200027

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Efficient visible/NIR light-driven uncaging of hydroxylated thiazole orange-based caged compounds in aqueous media2022

    • Author(s)
      Hashimoto Ryu、Minoshima Masafumi、Sakata Souhei、Ono Fumihito、Ishii Hirokazu、Watakabe Yuki、Nemoto Tomomi、Yanaka Saeko、Kato Koichi、Kikuchi Kazuya
    • Journal Title

      Chemical Science

      Volume: 13 Issue: 25 Pages: 7462-7467

    • DOI

      10.1039/d2sc02364d

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Photoinitiator‐Free Two‐Photon Polymerization of Biocompatible Materials for 3D Micro/Nanofabrication2022

    • Author(s)
      Nakayama Atsushi、Kumamoto Yasuaki、Minoshima Masafumi、Kikuchi Kazuya、Taguchi Atsushi、Fujita Katsumasa
    • Journal Title

      Advanced Optical Materials

      Volume: 10 Issue: 18 Pages: 2200474-2200474

    • DOI

      10.1002/adom.202200474

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] カテプシンK活性の検出を目指したOff-Onスイッチング19F MRIナノプローブの開発2022

    • Author(s)
      小西祐輝、杉原文徳、蓑島維文、菊地和也
    • Journal Title

      日本分子イメージング学会機関誌

      Volume: 15 Pages: 29-33

    • Related Report
      2022 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Cardioluminescence in Transgenic Zebrafish Larvae: A Calcium Imaging Tool to Study Drug Effects and Pathological Modeling2021

    • Author(s)
      Vicente Manuel、Salgado-Almario Jussep、Collins Michelle M.、Martinez-Sielva Antonio、Minoshima Masafumi、Kikuchi Kazuya、Domingo Beatriz、Llopis Juan
    • Journal Title

      Biomedicines

      Volume: 9 Issue: 10 Pages: 1294-1294

    • DOI

      10.3390/biomedicines9101294

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Anti-Siglec-15 antibody suppresses bone resorption by inhibiting osteoclast multinucleation without attenuating bone formation2021

    • Author(s)
      Tsukazaki Hiroyuki、Kikuta Junichi、Ao Tomoka、Morimoto Akito、Fukuda Chie、Tsuda Eisuke、Minoshima Masafumi、Kikuchi Kazuya、Kaito Takashi、Ishii Masaru
    • Journal Title

      Bone

      Volume: 152 Pages: 116095-116095

    • DOI

      10.1016/j.bone.2021.116095

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Development of Off-On Switching 19F MRI Probes for Cathepsin K Activity Detection2021

    • Author(s)
      Konishi Yuki、Okunishi Atsuya、Sugihara Fuminori、Nakamura Tatsuya、Akazawa Kazuki、Minoshima Masafumi、Kikuchi Kazuya
    • Journal Title

      Bulletin of the Chemical Society of Japan

      Volume: 94 Issue: 6 Pages: 1690-1694

    • DOI

      10.1246/bcsj.20210099

    • NAID

      130008066557

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Near-infrared fluorescent probes: a next-generation tool for protein-labeling applications2021

    • Author(s)
      Reja Shahi Imam、Minoshima Masafumi、Hori Yuichiro、Kikuchi Kazuya
    • Journal Title

      Chemical Science

      Volume: 12 Issue: 10 Pages: 3437-3447

    • DOI

      10.1039/d0sc04792a

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Optical Manipulation of Subcellular Protein Translocation Using a Photoactivatable Covalent Labeling System2021

    • Author(s)
      Kowada Toshiyuki、Arai Keisuke、Yoshimura Akimasa、Matsui Toshitaka、Kikuchi Kazuya、Mizukami Shin
    • Journal Title

      Angewandte Chemie International Edition

      Volume: 60 Issue: 20 Pages: 11378-11383

    • DOI

      10.1002/anie.202016684

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Fluorogenic probes for detecting deacylase and demethylase activity towards post-translationally-modified lysine residues2021

    • Author(s)
      Hori Yuichiro、Nishiura Miyako、Tao Tomomi、Baba Reisuke、Bull Steven D.、Kikuchi Kazuya
    • Journal Title

      Chemical Science

      Volume: 12 Issue: 7 Pages: 2498-2503

    • DOI

      10.1039/d0sc06551j

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Presentation] Development of 19F Magnetic Resonance Imaging Probe for in vivo Sensing Biomarkers2023

    • Author(s)
      Kazuya Kikuchi
    • Organizer
      ICBIC-20
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 天然物が織り成す化合物潜在空間が拓く生物活性分子デザイン2023

    • Author(s)
      菊地和也
    • Organizer
      「AIと有機合成化学」第11回勉強会
    • Related Report
      2023 Annual Research Report
    • Invited
  • [Presentation] Development of 19F Magnetic Resonance Imaging Probe for in vivo Sensing Biomarkers2023

    • Author(s)
      Kazuya Kikuchi
    • Organizer
      16th International Symposium on Applied Bioinorganic Chemistry (ISABC16)
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Chemical Probes with Tunable Fluorescence Properties2023

    • Author(s)
      Kazuya Kikuchi
    • Organizer
      FASEB Science Research Conference
    • Related Report
      2023 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Nano-particle Based 19F MRI Contrast Agent with Tunable Chemical Switches2022

    • Author(s)
      Kazuya Kikuchi
    • Organizer
      The Future of Molecular MR 2022
    • Related Report
      2022 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Nano-particle based 19F MRI contrast agent with tunable chemical switches2021

    • Author(s)
      Kazuya Kikuchi
    • Organizer
      Pacifichem 2021
    • Related Report
      2021 Annual Research Report
    • Int'l Joint Research / Invited
  • [Book] 先端の分析法 第2版(原理編第3章プローブ, MRIプローブ担当)2022

    • Author(s)
      澤田 嗣郎、小澤 岳昌、北森 武彦、中村 洋(東京理科大学名誉教授)、藤浪 眞紀、宮村 一夫、石丸 洋一郎、浦野 泰照、加地 範匡、坂 真智子、鈴木 茂、瀬藤 光利、宗林 由樹、馬場 嘉信、船津 公人、本田 暁紀、末永 智一、宮野 博、本山 晃
    • Total Pages
      1072
    • Publisher
      エヌ・ティー・エス
    • ISBN
      9784860437374
    • Related Report
      2021 Annual Research Report
  • [Patent(Industrial Property Rights)] タンパク質間相互作用を可視化する方法2024

    • Inventor(s)
      蓑島維文、Jiatong Li、菊地和也
    • Industrial Property Rights Holder
      蓑島維文、Jiatong Li、菊地和也
    • Industrial Property Rights Type
      特許
    • Filing Date
      2024
    • Related Report
      2023 Annual Research Report
  • [Patent(Industrial Property Rights)] タンパク質を蛍光標識する手法2023

    • Inventor(s)
      蓑島維文、Reja Shahi Imam、飯嶋航平、菊地和也
    • Industrial Property Rights Holder
      蓑島維文、Reja Shahi Imam、飯嶋航平、菊地和也
    • Industrial Property Rights Type
      特許
    • Filing Date
      2023
    • Related Report
      2022 Annual Research Report
  • [Patent(Industrial Property Rights)] ケージド化合物から化合物を放出させる方法2021

    • Inventor(s)
      蓑島維文、橋本 龍、菊地和也
    • Industrial Property Rights Holder
      蓑島維文、橋本 龍、菊地和也
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2021-022014
    • Filing Date
      2021
    • Related Report
      2021 Annual Research Report

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Published: 2021-04-28   Modified: 2025-01-30  

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