Design, Synthesis, and Biological Application of in vivo Imaging Probes

• Project/Area Number: 25220207
• Research Category: Grant-in-Aid for Scientific Research (S)
• Allocation Type: Single-year Grants
• Research Field: Chemical biology
• Research Institution: Osaka University
• Principal Investigator: Kikuchi Kazuya 大阪大学, 工学研究科, 教授 (70292951)
• Research Collaborator: HORI Yuichiro ; SUGIHARA Fuminori ; MINOSHIMA Masafumi
• Project Period (FY): 2013-05-31 – 2018-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥218,920,000 (Direct Cost: ¥168,400,000、Indirect Cost: ¥50,520,000); Fiscal Year 2017: ¥27,040,000 (Direct Cost: ¥20,800,000、Indirect Cost: ¥6,240,000); Fiscal Year 2016: ¥27,560,000 (Direct Cost: ¥21,200,000、Indirect Cost: ¥6,360,000); Fiscal Year 2015: ¥30,420,000 (Direct Cost: ¥23,400,000、Indirect Cost: ¥7,020,000); Fiscal Year 2014: ¥65,520,000 (Direct Cost: ¥50,400,000、Indirect Cost: ¥15,120,000); Fiscal Year 2013: ¥68,380,000 (Direct Cost: ¥52,600,000、Indirect Cost: ¥15,780,000)
• Keywords: in vivoイメージング / 化学プローブ / 分子イメージング / MRI / 破骨細胞 / in vivo イメージング / 蛋白質ラベル化 / 19FMRI / 19F MRIプローブ / オートファジー / BLタグ

Outline of Final Research Achievements

In this study, we designed and synthesized functional chemical probes for visualization of physiological phenomena in living animals. (1) We developed highly sensitive 19F MRI nanoprobes composed of perfluorocarbon-encapsulated silica nanoparticles. We used the nanoprobes for visualization of tumor tissue, detection of enzyme activity, and multicolor imaging. (2) We developed pH-responsive fluorescent probes for imaging acidic pH regions in bone tissue during osteoclastic bone resorption. The real-time images revealed actual behaviors of activated osteoclasts in living mice.
These functional molecular probes provide new tools for analyzing dynamics and function of biomolecules in vivo.

Academic Significance and Societal Importance of the Research Achievements

本研究で開発した化学プローブは、生きた動物個体内で起こる細胞や分子の機能を理解するための有用なツールとなりうる。特に、19F MRIを利用した生体深部における酵素活性の検出や、破骨細胞によって骨が溶解する様子は本研究で開発した化学プローブによって初めて観察できたものである。これらの化学プローブを用いて、動物個体内での生理機能や薬剤に対する応答の詳細な解析が今後期待できる。

Assessment Rating

Verification Result (Rating)

A

Research Products

• [Journal Article] Highly Sensitive Detection of Caspase-3/7 Activity in Living Mice Using Enzyme-Responsive 19F MRI Nanoprobes (2018)
  • Author(s): Akazawa Kazuki、Sugihara Fuminori、Nakamura Tatsuya、Mizukami Shin、Kikuchi Kazuya
  • Journal Title: Bioconjugate Chemistry Volume: 29 Issue: 5 Pages: 1720-1728
  • DOI: 10.1021/acs.bioconjchem.8b00167
  • Peer Reviewed
• [Journal Article] Sensing caspase-1 activity using activatable 19F MRI nanoprobes with improved turn-on kinetics (2018)
  • Author(s): Akazawa Kazuki、Sugihara Fuminori、Minoshima Masafumi、Mizukami Shin、Kikuchi Kazuya
  • Journal Title: Chemical Communications Volume: 54 Issue: 83 Pages: 11785-11788
  • DOI: 10.1039/c8cc05381b
  • Peer Reviewed
• [Journal Article] Perfluorocarbon-based 19F MRI Nanoprobes for In Vivo Multicolor Imaging (2018)
  • Author(s): Akazawa Kazuki、Sugihara Fuminori、Nakamura Tatsuya、Matsushita Hisashi, Mukai Hiroaki, Akimoto Rena, Minoshima Masafumi, Mizukami Shin、Kikuchi Kazuya
  • Journal Title: Angewante Chemie International Edition Volume: 57 Pages: 16742-16747
  • Peer Reviewed / Open Access
• [Journal Article] Synthetic-Molecule/Protein Hybrid Probe with Fluorogenic Switch for Live-Cell Imaging of DNA Methylation (2018)
  • Author(s): Yuichiro Hori, Norimichi Otomura, Ayuko Nishida, Miyako Nishiura, Maho Umeno, Isao Suetake, Kazuya Kikuchi
  • Journal Title: J Am Chem Soc Volume: 140 Issue: 5 Pages: 1686-1690
  • DOI: 10.1021/jacs.7b09713
  • Peer Reviewed / Open Access
• [Journal Article] Ratiometric Imaging of Intracellular Mg2+ Dynamics Using a Red Fluorescent Turn-off Probe and a Green Fluorescent Turn-on Probe (2018)
  • Author(s): Matsui Yusuke、Mizukami Shin、Kikuchi Kazuya
  • Journal Title: Chemistry Letters Volume: 47 Issue: 1 Pages: 23-26
  • DOI: 10.1246/cl.170918
  • NAID: 130006302062
  • Peer Reviewed
• [Journal Article] Intracellular Protein-Labeling Probes for Multicolor Single-Molecule Imaging of Immune Receptor?Adaptor Molecular Dynamics (2017)
  • Author(s): Sato Ryota、Kozuka Jun、Ueda Masahiro、Mishima Reiko、Kumagai Yutaro、Yoshimura Akimasa、Minoshima Masafumi、Mizukami Shin、Kikuchi Kazuya
  • Journal Title: Journal of the American Chemical Society Volume: 139 Issue: 48 Pages: 17397-17404
  • DOI: 10.1021/jacs.7b08262
  • Peer Reviewed
• [Journal Article] Visualization of long-term Mg2+ dynamics in apoptotic cells using a novel targetable fluorescent probe (2017)
  • Author(s): Matsui Y, Funato Y, Imamura H, Miki H, Mizukami S, Kikuchi K
  • Journal Title: Chem Sci. Volume: 8 Issue: 12 Pages: 8255-8264
  • DOI: 10.1039/c7sc03954a
  • Peer Reviewed / Open Access
• [Journal Article] Highly selective tridentate fluorescent probes for visualizing intracellular Mg2+ dynamics without interference from Ca2+ fluctuation (2017)
  • Author(s): Matsui Yusuke、Sadhu Kalyan K.、Mizukami Shin、Kikuchi Kazuya
  • Journal Title: Chemical Communications Volume: 53 Issue: 77 Pages: 10644-10647
  • DOI: 10.1039/c7cc06141b
  • Peer Reviewed
• [Journal Article] Photostable and Photoswitching Fluorescent Dyes for Super-resolution Imaging (2017)
  • Author(s): M. Minoshima, K. Kikuchi
  • Journal Title: Journal of Biological Inorganic Chemistry Volume: - Issue: 5 Pages: 1-14
  • DOI: 10.1007/s00775-016-1435-y
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Enzyme-Triggered Compound Release using Functionalized Antimicrobial Peptide Derivatives (2017)
  • Author(s): Shin Mizukami, Masayoshi Kashibe, Kengo Matsumoto, Yuichiro Hori, Kazuya Kikuchi
  • Journal Title: Chem. Sci. Volume: 8 Issue: 4 Pages: 3047-3053
  • DOI: 10.1039/c6sc04435b
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Fluorogenic Probes Reveal a Role of GLUT4 N-glycosylation in Intracellular Trafficking (2016)
  • Author(s): S. Hirayama, Y. Hori, Z. Benedek, T. Suzuki, K. Kikuchi
  • Journal Title: Nature Chemical Biology Volume: 12 Issue: 10 Pages: 853-859
  • DOI: 10.1038/nchembio.2156
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Real-Time Intravital Imaging of pH Variation Associated with Osteoclast Activity (2016)
  • Author(s): Hiroki Maeda, Toshiyuki Kowada, Junichi Kikuta, Masayuki Furuya, Mai Shirazaki, Shin Mizukami, Masaru Ishii, Kazuya Kikuchi
  • Journal Title: Nat. Chem. Biol. Volume: 12 Issue: 8 Pages: 579-585
  • DOI: 10.1038/nchembio.2096
  • Peer Reviewed
• [Journal Article] Fabrication of ‘Clickable’ Polyfluorene Nanowires with High Aspect Ratio as Biological Sensing Platforms (2016)
  • Author(s): T. Wasin, K. Enomoto, T. Sakurai, V. Padalkar, H. Cheng, M. Tang, A. Horio, D., Sakamaki, M. Omichi, A. Saeki, K. Kikuchi, Y. Hori, A. Chiba, Y. Saito, T. Kamiya, M. Sugimoto, S. Seki
  • Journal Title: ACS Sensors Volume: 6 Issue: 6 Pages: 766-774
  • DOI: 10.1021/acssensors.6b00070
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Design of a Protein tag and Fluorogenic Probe with Modular Structure for Live-Cell Imaging of Intracellular Proteins (2016)
  • Author(s): Y. Kamikawa, Y. Hori, K. Yamashita, L. Jin, S. Hirayama, D.M. Standley & K. Kikuchi
  • Journal Title: Chemical Science Volume: 7 Issue: 1 Pages: 308-314
  • DOI: 10.1039/c5sc02351c
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Real-time Intravital Imaging of pH Variation Associated with Osteoclast Activity (2016)
  • Author(s): H. Maeda, T. Kowada, J. Kikuta, M. Furuya, M. Shirazaki, S. Mizukami, M, Ishii & K. Kikuchi
  • Journal Title: Nature Chemical Biology Volume: 12
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Redesign of Fluorogenic Labeling System to Improve Surface Charges, Brightness, and Binding Kinetics for Imaging Functional Localization of Bromodomains (2015)
  • Author(s): Y. Hori, S. Hirayama, M. Sato & K. Kikuchi
  • Journal Title: Angewante Chemie International Edition Volume: 54 Issue: 48 Pages: 14368-14371
  • DOI: 10.1002/anie.201506935
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Actavataqble 19F MRI nanoparticle probes for the detection of reducing environments (2015)
  • Author(s): Nkamura, T., Matsushita, H., Sugihara, F., Yoshioka, Y., Mizukami, S. and Kikuchi, K.
  • Journal Title: Angewandte Chemi International Edition Volume: 54 Issue: 3 Pages: 1007-1010
  • DOI: 10.1002/anie.201409365
  • Peer Reviewed / Open Access
• [Journal Article] An Enzyme-Responsive Metal-Enhanced Near-Infrared Fluorescence Sensor Based on Functionalized Gold Nanoparticles (2015)
  • Author(s): Zhanghua Zeng, Shin Mizukami, Katsumasa Fujita, Kazuya Kikuchi
  • Journal Title: Chem. Sci. Volume: 6 Issue: 8 Pages: 4934-4939
  • DOI: 10.1039/c5sc01850a
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] 19F MRI Probes with Tunable Switches and Highly Sensitive 19F MRI Nano-probes (2015)
  • Author(s): K. Kikuchi
  • Journal Title: Bulletein of the Chemical Society of Japan Volume: 88 Issue: 4 Pages: 518-522
  • DOI: 10.1246/bcsj.20140392
  • NAID: 130005064990
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] BODIPY-based probes for the fluorescence imaging of biomolecules in living cells (2015)
  • Author(s): Toshiyuki Kowada, Hiroki Maeda, Kazuya Kikuchi
  • Journal Title: Chemical Society Reviews Volume: - Issue: 14 Pages: 4953-4972
  • DOI: 10.1039/c5cs00030k
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Intramolecular Long-distance Nucleophilic Reactions as a Rapid Fluorogenic Switch Applicable to Detection of Enzymatic Activity (2015)
  • Author(s): Reisuke BABA, Yuichiro HORI, Kazuya KIKUCHI
  • Journal Title: Chem. Eur. J. Volume: 21 Issue: 12 Pages: 4695-4702
  • DOI: 10.1002/chem.201406093
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Development of a Fluorogenic Probe Based on a DNA Staining Dye for Continuous Monitoring of the Histone Deacetylase Reaction (2014)
  • Author(s): M. Minoshima, T. Matsumoto, and K. Kikuchi
  • Journal Title: Anal. Chem. Volume: 86 Issue: 15 Pages: 7925-7930
  • DOI: 10.1021/ac501881s
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] New Biological Findings Which Were Revealed by Designed Chemical Probes (2018)
  • Author(s): Kikuchi Kazuya
  • Organizer: A3 Meeting for Chemical Biology
  • Int'l Joint Research / Invited
• [Presentation] In vivo Chemical Probes for MRI and Fluorescence Imaging (2018)
  • Author(s): Kikuchi Kazuya
  • Organizer: The 9th Asian Biological Inorganic Chemistry Conference (AsBIC IX)
  • Int'l Joint Research / Invited
• [Presentation] Development of Multifunctional 19F MRI Contrast Agents Based on Fluorine-encapsulated Silica Nanoparticle (2018)
  • Author(s): Kikuchi Kazuya
  • Organizer: Imaging in 2020
  • Int'l Joint Research / Invited
• [Presentation] New Biological Findings Which Were Clarified by Designed Chemical Probes (2018)
  • Author(s): Kikuchi Kazuya
  • Organizer: MSMLG 2018, 6th International Conference on Molecular Sensors and Molecular Logic Gates
  • Int'l Joint Research / Invited
• [Presentation] Real-time Intravital Imaging of pH Variation Associated with Osteoclast Activity Using BODIPY Based Two Photon Excitation Probes (2017)
  • Author(s): Kikuchi Kazuya
  • Organizer: 14th International Symposium on Applied Bioinorganic Chemistry
  • Int'l Joint Research / Invited
• [Presentation] Real-time Intravital Imaging of pH Variation Associated with Osteoclast Activity Using BODIPY Based Two Photon Excitation Probes (2016)
  • Author(s): K. Kikuchi
  • Organizer: The 8th Asian Biological Inorganic Chemistry Conference (AsBIC VIII)
  • Place of Presentation: Auckland, New Zealand
  • Year and Date: 2016-12-05
  • Int'l Joint Research / Invited
• [Presentation] Real-time Intravital Imaging of pH Variation Associated with Osteoclast Activity (2016)
  • Author(s): K. Kikuchi
  • Organizer: MSMLG 2016, 5th International Conference on Molecular Sensors and Molecular Logic Gates
  • Place of Presentation: Bath, UK
  • Year and Date: 2016-07-24
  • Int'l Joint Research / Invited
• [Presentation] Real-time Intravital Imaging of pH Variation Associated with Osteoclast Activity Using BODIPY Based Two Photon Excitation Probes (2016)
  • Author(s): K. Kikuchi
  • Organizer: ICPP-9, 9th International Conference on Porphyrins and Phthalocyanines
  • Place of Presentation: Nanjing, China
  • Year and Date: 2016-07-03
  • Int'l Joint Research / Invited
• [Presentation] Development of Multifunctional 19F MRI Contrast Agents with Fluorine-encapsulated Silica Nanoparticle (2015)
  • Author(s): K. Kikuchi
  • Organizer: Pacifichem2015
  • Place of Presentation: Honolulu HI, USA
  • Year and Date: 2015-12-15
  • Int'l Joint Research / Invited
• [Presentation] Development of Multifunctional 19F MRI Contrast Agents with Fluorine-encapsulated Silica Nanoparticle (2015)
  • Author(s): Kazuya Kikuchi
  • Organizer: Pacifichem2015
  • Place of Presentation: Honolulu, HI, USA
  • Year and Date: 2015-12-15
  • Int'l Joint Research / Invited
• [Presentation] Highly Sensitive 19F MRI Probes with Tunable Chemical Switches Using Metal Complexes (2015)
  • Author(s): K. Kikuchi
  • Organizer: ICBIC-17, International Conference on Biological Inorganic Chemistry
  • Place of Presentation: Beijing, China
  • Year and Date: 2015-07-20
  • Int'l Joint Research / Invited
• [Presentation] Development of Multifunctional 19F MRI Contrast Agents with Fluorine-encapsulated Silica Nanoparticle (2015)
  • Author(s): K. Kikuchi
  • Organizer: 13th International Symposium on Applied Bioinorganic Chemistry (ISABC13)
  • Place of Presentation: Galway, Ireland
  • Year and Date: 2015-06-22
  • Int'l Joint Research / Invited
• [Presentation] Intracellular Protein Labeling by Functional Probes with Tunable Chemical Switches (2014)
  • Author(s): Kazuya Kikuchi
  • Organizer: Labeling and Nanoscopy
  • Place of Presentation: Heidelberg, Gemany
  • Year and Date: 2014-09-24 – 2014-09-26
  • Invited
• [Patent(Industrial Property Rights)] メチル化DNAを蛍光標識する方法 (2015)
  • Inventor(s): 菊地和也、堀雄一郎、乙村法道
  • Industrial Property Rights Holder: 菊地和也、堀雄一郎、乙村法道
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-08-01