Regulation of invasive behavior by vascular endothelial growth factor is HEF1-dependent

Abstract

We previously reported a vascular endothelial growth factor (VEGF) autocrine loop in head and neck squamous cell carcinoma (HNSCC) cell lines, supporting a role for VEGF in HNSCC tumorigenesis. Using a phosphotyrosine proteomics approach, we screened the HNSCC cell line, squamous cell carcinoma-9 for effectors of VEGFR2 signaling. A cluster of proteins involved in cell migration and invasion, including the p130Cas paralog, human enhancer of filamentation 1 (HEF1/Cas-L/Nedd9) was identified. HEF1 silencing and overexpression studies revealed a role for VEGF in regulating cell migration, invasion and matrix metalloproteinase (MMP) expression in a HEF1-dependent manner. Moreover, cells plated on extracellular matrix-coated coverslips showed enhanced invadopodia formation in response to VEGF that was HEF1-dependent. Immunolocalization revealed that HEF1 colocalized to invadopodia with MT1-MMP. Analysis of HNSCC tissue microarrays for HEF1 immunoreactivity revealed a 6.5-fold increase in the odds of having a metastasis with a high HEF1 score compared with a low HEF1 score. These findings suggest that HEF1 may be prognostic for advanced stage HNSCC. They also show for the first time that HEF1 is required for VEGFmediated HNSCC cell migration and invasion, consistent with HEF1's recent identification as a metastatic regulator. These results support a strategy targeting VEGF:VEGFR2 in HNSCC therapeutics.

Integrin signalling adaptors: not only figurants in the cancer story

“…Indeed its expression levels is elevated in human metastatic melanoma compared with primary melanoma 18,19 and in invasive HNSCC 20 . Recently, a role in mammary tumorigenesis has also been proposed for NEDD9, whose absence significantly impairs tumour formation induced by the polyoma virus middle T oncogene (PyMT) 21 (Table II).…”

NSP-CAS Protein Complexes: Emerging Signaling Modules in Cancer

“…12,16,17,65 Interestingly, a distinctive role of NEDD9 in driving invasiveness and metastasis has been extensively documented in melanoma, breast cancer, and other cancers. 16,17,64,[66][67][68][69][70][71][72][73] In addition, analysis of NEDD9 knockout mice has implicated this CAS family member in tumor initiation in the MMTV-Polyoma middle T mouse mammary tumor model through activation of FAK, SRC, AKT, and ERK. 74 However, besides reducing tumor development, over time, the lack of NEDD9 expression can also promote cancer cell aggressiveness by causing genetic instability.…”

Signaling inputs to invadopodia and podosomes

“…In cancer cells, epidermal growth factor (EGF) is a well-described inducer of invadopodia formation (Mader et al, 2011;Yamaguchi et al, 2005). TGF-b (Mandal et al, 2008;Pignatelli et al, 2012), heparin binding (HB)-EGF (Díaz et al, 2013;Hayes et al, 2012), VEGF (Lucas et al, 2010) and hepatocyte growth factor/scatter factor (HGF) (Rajadurai et al, 2012) increase invadopodia numbers, whereas stromal cell derived factor 1a (SDF1a) has been shown to increase ECM degradation by breast cancer cells (Smith-Pearson et al, 2010). Many of these growth factor pathways converge on common signaling hubs, especially Src (1) Invadopodia/ podosome initiation.…”

“…(ii) MT1-MMP also cleaves other proteinases such as MMP2 (Sato et al, 1994). MMP2 is reported to disrupt vascular endothelial growth factor (VEGF)-heparin affin regulatory peptide (HARP) and VEGF-connective tissue growth factor (CTGF) angiogenic inhibitory complexes, which might release VEGF (Dean et al, 2007) and then trigger invadosome initiation (Lucas et al, 2010). Finally, MT1-MMP activation of PDGFR signaling (Lehti et al, 2005) is another potential mechanism that could promote invadosome formation (Eckert et al, 2011). kinase, phosphoinositide 3-kinases (PI3Ks) and Rho family GTPases, which ultimately control invadopodia and podosomes (Linder and Aepfelbacher, 2003;Murphy and Courtneidge, 2011).…”