Restriction of Receptor Movement Alters Cellular Response: Physical Force Sensing by EphA2

Moving Signals Many types of human breast cancers overexpress a cell-surface receptor—EphA2—a tyrosine kinase activated by the ligand ephrin-A1 present on adjoining cells. Salaita et al. (p. 1380; see the Perspective by Paszek and Weaver) studied the regulation of mechanically stimulated EphA2 signaling by inducing intermembrane signaling between living EphA2-expressing human breast cancer cells and supported membranes displaying laterally mobile ephrin-A1. When the receptors engaged their ligands, they formed clusters that moved radially to the junction between the cells and the membranes. Physically impeding this movement altered the cellular response to ephrin-A1. Different breast cancer cell lines showed differences in receptor movement that correlated with their invasion potential, and might indicate their capacity for metastasis formation. Mechanical forces acting on a cell-surface receptor affect the activation of a signaling pathway involved in breast cancer. Activation of the EphA2 receptor tyrosine kinase by ephrin-A1 ligands presented on apposed cell surfaces plays important roles in development and exhibits poorly understood functional alterations in cancer. We reconstituted this intermembrane signaling geometry between live EphA2-expressing human breast cancer cells and supported membranes displaying laterally mobile ephrin-A1. Receptor-ligand binding, clustering, and subsequent lateral transport within this junction were observed. EphA2 transport can be blocked by physical barriers nanofabricated onto the underlying substrate. This physical reorganization of EphA2 alters the cellular response to ephrin-A1, as observed by changes in cytoskeleton morphology and recruitment of a disintegrin and metalloprotease 10. Quantitative analysis of receptor-ligand spatial organization across a library of 26 mammary epithelial cell lines reveals characteristic differences that strongly correlate with invasion potential. These observations reveal a mechanism for spatio-mechanical regulation of EphA2 signaling pathways.