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A kinome-wide high-content siRNA screen identifies MEK5–ERK5 signaling as critical for breast cancer cell EMT and metastasis

Oncogene volume 37, pages 4197–4213 (2018)Cite this article

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Abstract

An epithelial to mesenchymal transition (EMT) has been correlated to malignant tumor progression and metastasis by promoting cancer cell migration and invasion and chemoresistance. Hence, finding druggable EMT effectors is critical to efficiently interfere with metastasis formation and to overcome therapy resistance. We have employed a high-content microscopy screen in combination with a kinome and phosphatome-wide siRNA library to identify signaling pathways underlying an EMT of murine mammary epithelial cells and breast cancer cells. This screen identified the MEK5–ERK5 axis as a critical player in TGFβ-mediated EMT. Suppression of MEK5–ERK5 signaling completely prevented the morphological and molecular changes occurring during a TGFβ-induced EMT and, conversely, forced highly metastatic breast cancer cells into a differentiated epithelial state. Inhibition of MEK5–ERK5 signaling also repressed breast cancer cell migration and invasion and substantially reduced lung metastasis without affecting primary tumor growth. The results suggest that the MEK5–ERK5 signaling axis via activation of MEF2B and other transcription factors plays an important role in the induction and maintenance of breast cancer cell migration and invasion and thus represents an exploitable target for the pharmacological inhibition of cancer cell metastasis.

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Acknowledgements

We thank P. Lorentz and the DBM microscopy facility (DBM, University of Basel) for excellent support with microscopy, H. Antoniadis, P. Schmidt, and I. Galm for technical support, and E. Fagiani for flow cytometry. We also thank the microscopy facility at Actelion Pharmaceutical, in particular Carina Lotz-Jenne, for input to the screening experiments, and F. Tang for help with phostag gels.

Funding

This work was supported by the SystemsX.ch MTD project MetastasiX, the Swiss National Science Foundation, the Swiss Cancer League, and Krebsliga Beider Basel. N.M.-S. was supported by a Marie-Heim Vögtlin grant from the Swiss National Foundation.

Author contributions

S.P. designed and performed the experiments, analyzed the data and wrote the paper. N.M.-S. helped in the screening set up, performed experiments and analyzed the data. M.D. performed experiments and analyzed data. M.S. isolated primary tumor cells. R.K.R.K. performed bioinformatics analysis. G.C. oversaw the project, designed experiments, analyzed data and wrote the paper.

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    1. Department of Biomedicine, University of Basel, Basel, 4058, Switzerland

      Simona Pavan, Nathalie Meyer-Schaller, Maren Diepenbruck, Ravi Kiran Reddy Kalathur, Meera Saxena & Gerhard Christofori

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    Pavan, S., Meyer-Schaller, N., Diepenbruck, M. et al. A kinome-wide high-content siRNA screen identifies MEK5–ERK5 signaling as critical for breast cancer cell EMT and metastasis. Oncogene 37, 4197–4213 (2018). https://doi.org/10.1038/s41388-018-0270-8

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