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Target-Specific Imaging of Cathepsin and S100A8/A9 Reflects Specific Features of Malignancy and Enables Estimation of Tumor Malignancy

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Abstract

Purpose

Tumor development and metastasis are dependent on tumor infiltrating immune cells which form a characteristic tumor microenvironment (TME). Activated monocytes secrete the protein heterodimer S100A8/A9 promoting TME formation. Monocyte-dependent proteases facilitate local tumor cell invasion by degradation of the extracellular matrix. We aimed for target specific in vivo imaging of S100A8 and proteases to provide differentiating biomarkers for local tumor growth and metastatic potential.

Procedures

Murine breast cancer cells of the 4T1 model with graduated metastatic potential (4T1 and 4T07: both hematogenous metastasis > 168FAR: lymph-node metastasis > 67NR: no metastasis) were orthotopically implanted into female BALB/c mice. At 4 mm size, tumors were investigated by injecting the protease-specific probe ProSense 750EX (PerkinElmer, 4T1 n = 7, 4T07 n = 10, 168FAR n = 16, 67NR n = 15) and anti-S100A8-Cy5.5 (n = 6 each) and performing fluorescence reflectance imaging at 0 and 24 h after injection. In vivo imaging was validated with immunohistochemistry.

Results

At 24 h, S100A8-specific signals in 4T1 and 4T07 were significantly higher (1714.05/1683.45 AU) as compared to 168FAR and 67NR (174.85/167.95 AU, p = 0.0012/p = 0.0003), reflecting the capability of hematogenous spread. Protease-specific signals were significantly higher in 4T1 and 4T07 (348.01/409.93 AU) as compared to 168FAR (214.91 AU) and 67NR (129.78 AU p < 0.0001 each), reflecting local vessel invasion and tumor cell shedding. Immunohistology supported the in vivo imaging results.

Conclusions

Non-invasive in vivo imaging of S100A8 and monocytic proteases allows for differentiation of the tumors’ local invasive and systemic metastatic potential in reflecting the TME formation. While proteases augment local tumor cell invasion, solid metastases seem to be dependent on a pro-tumoral microenvironment.

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Acknowledgments

We thank Heike Berheide, Klaudia Niepagenkemper, Claudia Terwesten-Solé, and Ina Winkler for excellent technical support.

Funding

This study was funded in part by the Interdisciplinary Center for Clinical Research of the University of Muenster (Vo2/014/09, PIX) and the German Research Foundation (DFG) CRC 1009 B8, B9, and EI 878 1-1.

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Author notes

  1. Anne Helfen and Nils Große Hokamp contributed equally to this work.

Authors and Affiliations

  1. Translational Research Imaging Center, Institute of Clinical Radiology, Medical Faculty, University of Muenster and University Hospital Muenster, Muenster, Germany

    Anne Helfen, Christiane Geyer, Walter Heindel, Carsten Höltke, Max Masthoff, Moritz Wildgruber & Michel Eisenblaetter

  2. Institute for Diagnostic and Interventional Radiology, University Hospital Cologne, Cologne, Germany

    Nils Große Hokamp

  3. Interdisciplinary Center for Clinical Research, University of Muenster, Muenster, Germany

    Christiane Geyer

  4. Department of Radiology, St Franziskus Hospital Muenster, Muenster, Germany

    Christoph Bremer

  5. Institute of Immunology, University Hospital Muenster, Muenster, Germany

    Thomas Vogl, Katarzyna Barczyk-Kahlert & Johannes Roth

  6. Department of Diagnostic and Interventional Radiology, Medical Center - University of Freiburg, Freiburg, Germany

    Michel Eisenblaetter

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  1. Anne Helfen
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Corresponding author

Correspondence to Michel Eisenblaetter.

Ethics declarations

All animal experiments in this study have been approved by the responsible authorities (Protocol No. 8.87-50.10.36.08.191).

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The authors declare that they have no conflict of interest.

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Helfen, A., Große Hokamp, N., Geyer, C. et al. Target-Specific Imaging of Cathepsin and S100A8/A9 Reflects Specific Features of Malignancy and Enables Estimation of Tumor Malignancy. Mol Imaging Biol 22, 66–72 (2020). https://doi.org/10.1007/s11307-019-01370-1

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  • DOI: https://doi.org/10.1007/s11307-019-01370-1

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