Technical Report | Published:

Selective molecular imaging of viable cancer cells with pH-activatable fluorescence probes

Nature Medicine volume 15, pages 104109 (2009) | Download Citation

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Abstract

A long-term goal of cancer diagnosis is to develop tumor-imaging techniques that have sufficient specificity and sensitivity. To achieve this goal, minimizing the background signal originating from nontarget tissues is crucial. Here we achieve highly specific in vivo cancer visualization by using a newly designed targeted 'activatable' fluorescent imaging probe. This agent is activated after cellular internalization by sensing the pH change in the lysosome. Novel acidic pH–activatable probes based on the boron-dipyrromethene fluorophore were synthesized and then conjugated to a cancer-targeting monoclonal antibody. As proof of concept, ex vivo and in vivo imaging of human epidermal growth factor receptor type 2–positive lung cancer cells in mice was performed. The probe was highly specific for tumors with minimal background signal. Furthermore, because the acidic pH in lysosomes is maintained by the energy-consuming proton pump, only viable cancer cells were successfully visualized. The design concept can be widely adapted to cancer-specific, cell surface–targeting molecules that result in cellular internalization.

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Acknowledgements

This study was supported in part by the Precursory Research for Embryonic Sciences and Technology from the Japan Science and Technology Agency, by research grants 19021010 and 19205021 to Y.U. and by the Intramural Research Program of the US National Institutes of Health, National Cancer Institute, Center for Cancer Research to Y.H., Y.K., T.B., P.L.C. and H.K.

Author information

Author notes

    • Yukihiro Hama
    • , Yoshinori Koyama
    •  & Tristan Barrett

    Present addresses: Department of Radiology, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama 359-8513, Japan (Y.H.); Department of Diagnostic and Interventional Radiology, Gunma University Hospital, 3-39-15 Showa-machi, Maebashi, Gunma 371-8511, Japan (Y.K.); Department of Radiology, Addenbrooke's Hospital, Cambridge University Teaching Hospitals National Health Service Foundation Trust, Hills Road, Cambridge, CB2 2QQ, UK (T.B.).

Affiliations

  1. Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-0033, Japan.

    • Yasuteru Urano
    • , Daisuke Asanuma
    • , Mako Kamiya
    •  & Tetsuo Nagano
  2. PRESTO, Japan Science and Technology Agency, 3-5 Sanbancho, Chiyoda, Tokyo 102-0075, Japan.

    • Yasuteru Urano
  3. Molecular Imaging Program, Center for Cancer Research, National Cancer Institute, US National Institutes of Health, Building 10, Room 1B40, Mail Stop Code 1088, 10 Center Drive, Bethesda, Maryland 20892-1088, USA.

    • Yukihiro Hama
    • , Yoshinori Koyama
    • , Tristan Barrett
    • , Peter L Choyke
    •  & Hisataka Kobayashi
  4. Molecular Diagnostic Technology Group, Advanced Core Technology Department, Research and Development Division, Olympus Corporation, 2-3 Kuboyama-cho, Hachioji, Tokyo 192-8512, Japan.

    • Toshiaki Watanabe
    •  & Akira Hasegawa

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Contributions

Y.U. and H.K. planned the projects, developed the probes, performed the in vivo experiments and wrote and edited the manuscript. D.A. and M.K. developed the probes, performed the in vivo experiments and wrote and edited the manuscript. Y.H., Y.K., T.B., T.W. and A.H. performed the in vivo experiments and wrote and edited the manuscript. T.N. and P.L.C. planned the projects and wrote and edited the manuscript.

Corresponding authors

Correspondence to Yasuteru Urano or Hisataka Kobayashi.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figs. 1–4, Supplementary Tables 1–3 and Supplementary Methods

Videos

  1. 1.

    Supplementary Video 1

    Reversible activation by altering the pH.

  2. 2.

    Supplementary Video 2

    Fluorescence-guided Laparoscope

  3. 3.

    Supplementary Video 3

    Fluorescence-guided Laparoscope

  4. 4.

    Supplementary Video 4

    Fluorescence-guided Laparoscope

  5. 5.

    Supplementary Video 5

    pH-activatable rhodamine

About this article

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DOI

https://doi.org/10.1038/nm.1854

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