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Ephrin-B2–EphB4 communication mediates tumor–endothelial cell interactions during hematogenous spread to spinal bone in a melanoma metastasis model

Oncogene volume 39, pages 7063–7075 (2020)Cite this article

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Abstract

Metastases account for the majority of cancer deaths. Bone represents one of the most common sites of distant metastases, and spinal bone metastasis is the most common source of neurological morbidity in cancer patients. During metastatic seeding of cancer cells, endothelial–tumor cell interactions govern extravasation to the bone and potentially represent one of the first points of action for antimetastatic treatment. The ephrin-B2–EphB4 pathway controls cellular interactions by inducing repulsive or adhesive properties, depending on forward or reverse signaling. Here, we report that in an in vivo metastatic melanoma model, ephrin-B2-mediated activation of EphB4 induces tumor cell repulsion from bone endothelium, translating in reduced spinal bone metastatic loci and improved neurological function. Selective ephrin-B2 depletion in endothelial cells or EphB4 inhibition increases bone metastasis and shortens the time window to hind-limb locomotion deficit from spinal cord compression. EphB4 overexpression in melanoma cells ameliorates the metastatic phenotype and improves neurological outcome. Timely harvesting of bone tissue after tumor cell injection and intravital bone microscopy revealed less tumor cells attached to ephrin-B2-positive endothelial cells. These results suggest that ephrin-B2–EphB4 communication influences bone metastasis formation by altering melanoma cell repulsion/adhesion to bone endothelial cells, and represents a molecular target for therapeutic intervention.

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Fig. 1: Endothelial ephrin-B2 controls spinal bone metastasis after retrograde carotid artery injection of melanoma cells.
Fig. 2: Effects of ephrin-B2–EphB4 signaling on spinal bone metastasis using LLC1 and EphB4-low melanoma cell lines.
Fig. 3: Intact forward and reverse signaling is required to maintain antimetastatic effects for ephrin-B2–EphB4 signaling.
Fig. 4: EphB4-overexpressing melanoma cells demonstrate reduced hematogenous metastatic spread to the spinal bone.

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Acknowledgements

Basic components of the cartoon are provided by Servier Medical Art. This work was supported by the German research foundation (DFG GEPRIS: 267716524) and the FOR2325 DFG Forschergruppe. TB was a doctoral student of the Charité Medical Neuroscience, NeuroCure cluster of excellence graduate school, received the Ernst von Leyden fellowship from the “Berliner Krebsgesellschaft e.V.,” and the Early/Advanced Postdoc Mobility fellowship from the Swiss National Science Foundation. MC was part of the Friedrich C. Luft Clinical Scientist Pilot Program funded by the Volkswagen Foundation and the Charité Foundation and the Clinical Fellow Program of the Berlin Institue of Health. The funding sources had no involvement in study design; in the collection, analysis, and interpretation of the data; in the writing of the report; in the decision to submit the paper for publication.

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Author notes

  1. These authors contributed equally: Peter Vajkoczy, Marcus Czabanka

Authors and Affiliations

  1. Department of Neurosurgery, Universitätsmedizin Charite, D-10117, Berlin, Germany

    Thomas Broggini, Andras Piffko, Adnan Ghori, Peter Vajkoczy & Marcus Czabanka

  2. Department of Physics, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA, 92093, USA

    Thomas Broggini

  3. Department of Neurosurgery, University Medical Center Hamburg-Eppendorf, Martinistraße 52, 20246, Hamburg, Germany

    Andras Piffko

  4. Department of Experimental Neurology, Center for Stroke Research Berlin, Universitätsmedizin Charite, D-10117, Berlin, Germany

    Christian J. Hoffmann & Christoph Harms

  5. Max-Planck-Institute for Molecular Biomedicine, Department of Tissue Morphogenesis, and University of Münster, Faculty of Medicine, D-48149, Münster, Germany

    Ralf H. Adams

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Correspondence to Marcus Czabanka.

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TB, AP, CJH, and AG performed experiments. TB, AP, and MC analyzed the data. CH and RHA provided materials. TB, PV, and MC designed experiments. TB, AP, PV, and MC wrote the paper. All authors read and approved the final paper.

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Broggini, T., Piffko, A., Hoffmann, C.J. et al. Ephrin-B2–EphB4 communication mediates tumor–endothelial cell interactions during hematogenous spread to spinal bone in a melanoma metastasis model. Oncogene 39, 7063–7075 (2020). https://doi.org/10.1038/s41388-020-01473-y

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