Non-canonical EphA2 activation underpins PTEN-mediated metastatic migration and poor clinical outcome in prostate cancer

Background The key process of mesenchymal to amoeboid transition (MAT), which enables prostate cancer (PCa) transendothelial migration and subsequent development of metastases in red bone marrow stroma, is driven by phosphorylation of EphA2 S897 by pAkt, which is induced by the omega-6 polyunsaturated fatty acid arachidonic acid. Here we investigate the influence of EphA2 signalling in PCa progression and long-term survival. Methods The mechanisms underpinning metastatic biopotential of altered EphA2 signalling in relation to PTEN status were assessed in vitro using canonical (EphA2 D739N ) and non-canonical (EphA2 S897G ) PC3-M mutants, interrogation of publicly available PTEN-stratified databases and clinical validation using a PCa TMA ( n = 177) with long-term follow-up data. Spatial heterogeneity of EphA2 was assessed using a radical prostatectomy cohort ( n = 67). Results Non-canonical EphA2 signalling via pEphA2 S897 is required for PCa transendothelial invasion of bone marrow endothelium. High expression of EphA2 or pEphA2 S897 in a PTEN low background is associated with poor overall survival. Expression of EphA2, pEphA2 S897 and the associated MAT marker pMLC2 are spatially regulated with the highest levels found within lesion areas within 500 µm of the prostate margin. Conclusion EphA2 MAT-related signalling confers transendothelial invasion. This is associated with a substantially worse prognosis in PTEN-deficient PCa.