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Deletion of ADAM-9 in HGF/CDK4 mice impairs melanoma development and metastasis

Oncogene volume 36, pages 5058–5067 (2017)Cite this article

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Abstract

ADAM-9 is a metalloproteinase expressed in peritumoral areas by invading melanoma cells and by adjacent peritumoral stromal cells; however, its function in stromal and melanoma cells is not fully understood. To address this question in vivo in a spontaneous melanoma model, we deleted ADAM-9 in mice carrying the hepatocyte growth factor (Hgf) transgene and knock-in mutation Cdk4R24C/R24C, demonstrated to spontaneously develop melanoma. Spontaneous melanoma arose less frequently in ADAM-9-deleted mice than in controls. Similarly reduced tumor numbers (although with faster growth kinetics) were detected upon induction of melanoma with 7,12-dimethylbenz[a]anthracene (DMBA). However, more lesions were induced at early time points in the absence of ADAM-9. Increased initial and decreased late tumor numbers were paralleled by altered tumor cell proliferation, but not apoptosis or inflammation. Importantly, significantly reduced lung metastases were detected upon ADAM-9 deletion. Using in vitro assays to address this effect mechanistically, we detected reduced adhesion and transmigration of ADAM-9-silenced melanoma cells to/through the endothelium. This implies that ADAM-9 functionally and cell autonomously mediates extravasation of melanoma cells. In vitro and in vivo we demonstrated that the basement membrane (BM) component laminin β3-chain is a direct substrate of ADAM-9, thus contributing to destabilization and disruption of the BM barrier during invasion. In in vitro invasion assays using human melanoma cells and skin equivalents, depletion of ADAM-9 resulted in decreased invasion of the BM, which remained almost completely intact, as shown by continuous staining for laminin β3-chain. Importantly, supplying soluble ADAM-9 to the system reversed this effect. Taken together, our data show that melanoma derived ADAM-9 autonomously contributes to melanoma progression by modulating cell adhesion to the endothelium and altering BM integrity by proteolytically processing the laminin-β3 chain. This newly described process and ADAM-9 itself may represent potential targets for anti-tumor therapies.

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Acknowledgements

We thank Prof Manuel Koch, Department of Biochemistry, University of Cologne for kindly providing the laminin β3 antibody and recombinant laminin β3-chain. We thank Jan Zamek, Claudia Coerper-Ochsman and Nina Ruers for excellent technical assistance. This work was supported by the Melanoma Research Network of the Deutsche Krebshilfe (Melanoma Verbund), by the Deutsche Forschungsgemeinschaft through the SFB829 (B4) and by Köln Fortune Program of the University of Cologne.

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Author notes

  1. N Giebeler and A Schönefuß: These authors equally contributed to this work.

Authors and Affiliations

  1. Department of Dermatology and Venerology, University of Cologne, Germany

    N Giebeler, A Schönefuß, C Mauch & P Zigrino

  2. Department of Dermatology and Allergology, University of Bonn, Germany

    J Landsberg

  3. Department of Dermatology, University of Magdeburg, Germany

    T Tüting

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  1. N Giebeler
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Correspondence to P Zigrino.

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Giebeler, N., Schönefuß, A., Landsberg, J. et al. Deletion of ADAM-9 in HGF/CDK4 mice impairs melanoma development and metastasis. Oncogene 36, 5058–5067 (2017). https://doi.org/10.1038/onc.2017.162

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