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Cell type-dependent pathogenic functions of overexpressed human cathepsin B in murine breast cancer progression

Oncogene volume 33, pages 4474–4484 (2014)Cite this article

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Abstract

The cysteine protease cathepsin B (CTSB) is frequently overexpressed in human breast cancer and correlated with a poor prognosis. Genetic deficiency or pharmacological inhibition of CTSB attenuates tumor growth, invasion and metastasis in mouse models of human cancers. CTSB is expressed in both cancer cells and cells of the tumor stroma, in particular in tumor-associated macrophages (TAM). In order to evaluate the impact of tumor- or stromal cell-derived CTSB on Polyoma Middle T (PyMT)-induced breast cancer progression, we used in vivo and in vitro approaches to induce human CTSB overexpression in PyMT cancer cells or stromal cells alone or in combination. Orthotopic transplantation experiments revealed that CTSB overexpression in cancer cells rather than in the stroma affects PyMT tumor progression. In 3D cultures, primary PyMT tumor cells showed higher extracellular matrix proteolysis and enhanced collective cell invasion when CTSB was overexpressed and proteolytically active. Coculture of PyMT cells with bone marrow-derived macrophages induced a TAM-like macrophage phenotype in vitro, and the presence of such M2-polarized macrophages in 3D cultures enhanced sprouting of tumor spheroids. We employed a doxycycline (DOX)-inducible CTSB expression system to selectively overexpress human CTSB either in cancer cells or in macrophages in 3D cocultures. Tumor spheroid invasiveness was only enhanced when CTSB was overexpressed in cancer cells, whereas CTSB expression in macrophages alone did not further promote invasiveness of tumor spheroids. We conclude that CTSB overexpression in the PyMT mouse model promotes tumor progression not by a stromal effect, but by a direct, cancer cell-inherent mode of action: CTSB overexpression renders the PyMT cancers more invasive by increasing proteolytic extracellular matrix protein degradation fostering collective cell invasion into adjacent tissue.

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Acknowledgements

The CTSB+/0 mice were kindly provided by Dr L. Pennacchio, Genomics Division, Lawrence Berkeley Laboratory, Berkeley, USA. The immortalized macrophage cell line from FVB/n mice was kindly provided by Eike Latz, currently University of Bonn, Germany. We thank Nicole Klemm, Anja Faulhaber and Julia Wolanski for excellent technical assistance. Proteolysis imaging was mainly performed with kind help of the Microscopy, Imaging & Cytometry Resources Core at Wayne State University, Detroit, MI, USA, directed by Dr Kamiar Moin and supported in part by NIH Center grant P30CA22453 to the Karmanos Cancer Institute, Wayne State University. This work was supported by the Excellence Initiative of the German Federal and State Governments (EXC 294 and GSC-4, Spemann Graduate School), and the Deutsche Forschungsgemeinschaft SFB 850 project B7.

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Author notes

  1. Current address: Memorial Sloan-Kettering Cancer Center, New York, USA.

Authors and Affiliations

  1. Institute of Molecular Medicine and Cell Research, Albert-Ludwigs-University Freiburg, Freiburg, Germany

    F Bengsch, A Buck, S C Günther, J R Seiz, M Tacke, L Sevenich, L E Hillebrand, U Kern, C Peters & T Reinheckel

  2. Spemann Graduate School of Biology and Medicine, Albert-Ludwigs-University Freiburg, Freiburg, Germany

    F Bengsch & U Kern

  3. Faculty of Biology, Albert-Ludwigs-University Freiburg, Freiburg, Germany

    F Bengsch, L E Hillebrand & U Kern

  4. Department of Hematology/Oncology, Genomics Core Lab, University Medical Center, Freiburg, Germany

    D Pfeifer

  5. Department of Radiology, Medical Physics, University Medical Center, Freiburg, Germany

    D von Elverfeldt

  6. BIOSS Centre for Biological Signalling Studies, Freiburg, Germany

    L E Hillebrand, C Peters & T Reinheckel

  7. Department of Pharmacology, Wayne State University, Detroit, MI, USA

    M Sameni & B F Sloane

  8. Barbara Ann Carmanos Cancer Institute, Wayne State University, Detroit, MI, USA

    B F Sloane

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Bengsch, F., Buck, A., Günther, S. et al. Cell type-dependent pathogenic functions of overexpressed human cathepsin B in murine breast cancer progression. Oncogene 33, 4474–4484 (2014). https://doi.org/10.1038/onc.2013.395

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