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Multispectral Photoacoustic Imaging of Tumor Protease Activity with a Gold Nanocage-Based Activatable Probe

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Abstract

Purpose

Tumor proteases have been recognized as significant regulators in the tumor microenvironment, but the current strategies for in vivo protease imaging have tended to focus on the development of a probe design rather than the investigation of a novel imaging strategy by leveraging the imaging technique and probe. Herein, it is the first report to investigate the ability of multispectral photoacoustic imaging (PAI) to estimate the distribution of protease cleavage sites inside living tumor tissue by using an activatable photoacoustic (PA) probe.

Procedures

The protease MMP-2 is selected as the target. In this probe, gold nanocages (GNCs) with an absorption peak at ~ 800 nm and fluorescent dye molecules with an absorption peak at ~ 680 nm are conjugated via a specific enzymatic peptide substrate. Upon enzymatic activation by MMP-2, the peptide substrate is cleaved and the chromophores are released. Due to the different retention speeds of large GNCs and small dye molecules, the probe alters its intrinsic absorption profile and produces a distinct change in the PA signal. A multispectral PAI technique that can distinguish different chromophores based on intrinsic PA spectral signatures is applied to estimate the signal composition changes and indicate the cleavage interaction sites. Finally, the multispectral PAI technique with the activatable probe is tested in solution, cultured cells, and a subcutaneous tumor model in vivo.

Results

Our experiment in solution with enzyme ± inhibitor, cell culture ± inhibitor, and in vivo tumor model with administration of the developed probe ± inhibitor demonstrated the probe was cleaved by the targeted enzyme. Particularly, the in vivo estimation of the cleavage site distribution was validated with the result of ex vivo immunohistochemistry analysis.

Conclusions

This novel synergy of the multispectral PAI technique and the activatable probe is a potential strategy for the distribution estimation of tumor protease activity in vivo.

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Acknowledgments

The authors acknowledge the use of the University Research Facility in Life Sciences (ULS).

Conflict of Interest

The authors declare that they have no conflict of interest.

Funding

This work was supported in part by the Hong Kong Innovation and Technology Fund (GHP/009/14SZ); Shenzhen Science, Technology and Innovation Commission research grant (SGLH20150216172854731); Health and Medical Research Fund of Food and Health Bureau of Hong Kong (01121836 and 03144266); National Natural Science Foundations of China (11674271); and The Hong Kong Polytechnic University internal grant (4-BCAP and 4-BCDP).

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Authors and Affiliations

  1. Department of Biomedical Engineering, The Hong Kong Polytechnic University, Hong Kong SAR, People’s Republic of China

    Cheng Liu, Shiying Li & Lei Sun

  2. Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong SAR, People’s Republic of China

    Yanjuan Gu & Wing-tak Wong

  3. Department of Ultrasound, The Second People’s Hospital of Shenzhen, Shenzhen, People’s Republic of China

    Huahua Xiong

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  1. Cheng Liu
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  2. Shiying Li
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  3. Yanjuan Gu
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  4. Huahua Xiong
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  6. Lei Sun
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Correspondence to Lei Sun.

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Liu, C., Li, S., Gu, Y. et al. Multispectral Photoacoustic Imaging of Tumor Protease Activity with a Gold Nanocage-Based Activatable Probe. Mol Imaging Biol 20, 919–929 (2018). https://doi.org/10.1007/s11307-018-1203-1

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