Endothelial Cell-Specific Molecule 2 (ECSM2) Localizes to Cell-Cell Junctions and Modulates bFGF-Directed Cell Migration via the ERK-FAK Pathway

Abstract

BackgroundDespite its first discovery by in silico cloning of novel endothelial cell-specific genes a decade ago, the biological functions of endothelial cell-specific molecule 2 (ECSM2) have only recently begun to be understood. Limited data suggest its involvement in cell migration and apoptosis. However, the underlying signaling mechanisms and novel functions of ECSM2 remain to be explored.Methodology/Principal FindingsA rabbit anti-ECSM2 monoclonal antibody (RabMAb) was generated and used to characterize the endogenous ECSM2 protein. Immunoblotting, immunoprecipitation, deglycosylation, immunostaining and confocal microscopy validated that endogenous ECSM2 is a plasma membrane glycoprotein preferentially expressed in vascular endothelial cells (ECs). Expression patterns of heterologously expressed and endogenous ECSM2 identified that ECSM2 was particularly concentrated at cell-cell contacts. Cell aggregation and transwell assays showed that ECSM2 promoted cell-cell adhesion and attenuated basic fibroblast growth factor (bFGF)-driven EC migration. Gain or loss of function assays by overexpression or knockdown of ECSM2 in ECs demonstrated that ECSM2 modulated bFGF-directed EC motility via the FGF receptor (FGFR)-extracellular regulated kinase (ERK)-focal adhesion kinase (FAK) pathway. The counterbalance between FAK tyrosine phosphorylation (activation) and ERK-dependent serine phosphorylation of FAK was critically involved. A model of how ECSM2 signals to impact bFGF/FGFR-driven EC migration was proposed.Conclusions/SignificanceECSM2 is likely a novel EC junctional protein. It can promote cell-cell adhesion and inhibit bFGF-mediated cell migration. Mechanistically, ECSM2 attenuates EC motility through the FGFR-ERK-FAK pathway. The findings suggest that ECSM2 could be a key player in coordinating receptor tyrosine kinase (RTK)-, integrin-, and EC junctional component-mediated signaling and may have important implications in disorders related to endothelial dysfunction and impaired EC junction signaling.

Endothelial Cell-specific Chemotaxis Receptor (ECSCR) Enhances Vascular Endothelial Growth Factor (VEGF) Receptor-2/Kinase Insert Domain Receptor (KDR) Activation and Promotes Proteolysis of Internalized KDR

“…16). For example, loss of ECSCR has resulted in differential impact on EC migration, depending on stimulus, cell type, and assay conditions (1,11,13,14). Our earlier (1) and current observations indicate that loss and gain of ECSCR can both result in decreased VEGF-dependent migration.…”

Favorable prognosis of operable non-small cell lung cancer (NSCLC) patients harboring an increased expression of tumor endothelial markers (TEMs)

“…Three of these identified genes are the magic roundabout receptor 4 (Robo4), C-type lectin 14 (Clec14) and the endothelial-cell-specific molecule 2 (ECSM2), synonymously named endothelial cell-specific chemotaxis regulator (ECSCR) [5,6]. In general, the Robo4 protein is a single pass transmembrane cell surface receptor and a member of the roundabout receptorfamily (Robo1-4) [7].…”

“…ECSCR is an endothelial cell junction protein also involved in the regulation, migration and apoptosis of endothelial cells. Functional down-regulation of the ECSCR protein in endothelial cells showed reduced chemotaxis and impaired tube formation on matrigel [5]. The biological role of ECSCR in tumor angiogenesis still remains to be explored.…”

The tumor suppressor geneARHI(DIRAS3)inhibits ovarian cancer cell migration through multiple mechanisms

“…[45][46][47] The activation of FAK may be further enhanced by the costimulation of growth factor receptors by ECM associated growth factors, such as bFGF, EGF or PDGF. [48][49][50] FAK is a nonreceptor protein tyrosine kinase (NRPTK) that is typically, but not always, associated with supramolecular focal adhesion (FA) complexes (FAC). Focal adhesion complex assembly and disassembly are critical for cell attachment and movement.…”