EpCAM ectodomain EpEX is a ligand of EGFR that counteracts EGF-mediated epithelial-mesenchymal transition through modulation of phospho-ERK1/2 in head and neck cancers

Abstract

Head and neck squamous cell carcinomas (HNSCCs) are characterized by outstanding molecular heterogeneity that results in severe therapy resistance and poor clinical outcome. Inter- and intratumoral heterogeneity in epithelial-mesenchymal transition (EMT) was recently revealed as a major parameter of poor clinical outcome. Here, we addressed the expression and function of the therapeutic target epidermal growth factor receptor (EGFR) and of the major determinant of epithelial differentiation epithelial cell adhesion molecule (EpCAM) in clinical samples and in vitro models of HNSCCs. We describe improved survival of EGFR ^low/EpCAM ^high HNSCC patients ( n = 180) and provide a molecular basis for the observed disparities in clinical outcome. EGF/EGFR have concentration-dependent dual capacities as inducers of proliferation and EMT through differential activation of the central molecular switch phosphorylated extracellular signal–regulated kinase 1/2 (pERK1/2) and EMT transcription factors (EMT-TFs) Snail, zinc finger E-box-binding homeobox 1 (Zeb1), and Slug. Furthermore, soluble ectodomain of EpCAM (EpEX) was identified as a ligand of EGFR that activates pERK1/2 and phosphorylated AKT (pAKT) and induces EGFR-dependent proliferation but represses EGF-mediated EMT, Snail, Zeb1, and Slug activation and cell migration. EMT repression by EpEX is realized through competitive modulation of pERK1/2 activation strength and inhibition of EMT-TFs, which is reflected in levels of pERK1/2 and its target Slug in clinical samples. Accordingly, high expression of pERK1/2 and/or Slug predicted poor outcome of HNSCCs. Hence, EpEX is a ligand of EGFR that induces proliferation but counteracts EMT mediated by the EGF/EGFR/pERK1/2 axis. Therefore, the emerging EGFR/EpCAM molecular cross talk represents a promising target to improve patient-tailored adjuvant treatment of HNSCCs.

Author summary

Head and neck squamous cell carcinomas (HNSCCs) display poor survival, with death rates above 55%. Major factors affecting survival are metastases’ formation and therapy resistance. Phenotypic changes during partial epithelial-mesenchymal transition (EMT) provide tumor cells with increased migration, invasion, and therapy resistance. Understanding molecular mechanisms of EMT, as a central process of the metastatic cascade and the development of therapy resistance, is therefore important. In the present work, we identified molecular cross talk between epidermal growth factor receptor (EGFR) and epithelial cell adhesion molecule (EpCAM) as a novel determinant of clinical outcome in HNSCCs. Low levels of EGFR but high levels of EpCAM (EGFR ^low/EpCAM ^high) were associated with favorable prognosis, with survival rates above 90%, whereas EGFR ^high/EpCAM ^low correlated with poor survival, below 10%. EGFR was shown to have a concentration-dependent capacity to induce proliferation and EMT. Proteolytic cleavage of the extracellular domain of EpCAM (EpEX) produces a ligand of EGFR that induces EGFR-dependent proliferation but counteracts EGF-induced EMT. We delineate an EGFR/extracellular signal–regulated kinase 1/2 (ERK1/2)/EpCAM signaling axis that may be a promising therapeutic target for HNSCCs.