WNT5A-ROR2 axis mediates VEGF dependence of BRAF mutant melanoma

Abstract

Purpose Despite recent advances, approximately 50% of patient with metastatic melanoma eventually succumb to the disease. Patients with melanomas harboring a BRAF mutation (BRAFMut) have a worse prognosis than those with wildtype (BRAFWT) tumors. Unexpectedly, interim AVAST-M Phase III trial data reported benefit from adjuvant anti-VEGF bevacizumab only in the BRAFMut group. We sought to find mechanisms underpinning this sensitivity. Methods We investigated this finding in vitro and in vivo using melanoma cell lines and clones generated by BRAFV600E knock-in on a BRAFWT background. Results Compared with BRAFWT cells, isogenic BRAFV600E clones secreted more VEGF and exhibited accelerated growth rates as spheroids and xenografts, which were more vascular and proliferative. Recapitulating AVAST-M findings, bevacizumab affected only BRAFV600E xenografts, inducing significant tumor growth delay, reduced vascularity and increased necrosis. We identified 814 differentially expressed genes in isogenic BRAFV600E/BRAFWT clones. Of 61 genes concordantly deregulated in clinical melanomas ROR2 was one of the most upregulated by BRAFV600E. ROR2 was shown to be RAF-MEK regulated in BRAFV600E cells and its depletion suppressed VEGF secretion down to BRAFWT levels. The ROR2 ligand WNT5A was also overexpressed in BRAFMut melanomas, and in ROR2-overexpressing BRAFV600E cells MEK inhibition downregulated WNT5A and VEGF secretion. Conclusions These data implicate WNT5A-ROR2 in VEGF secretion, vascularity, adverse outcomes and bevacizumab sensitivity of BRAFMut melanomas, suggesting that this axis has potential therapeutic relevance.

Role and Function of Receptor Tyrosine Kinases in BRAF Mutant Cancers

“…The establishment of BRAF V600E mutation in melanomas was associated with upregulated WNT5A expression leading to increased VEGF secretion and vascularization of melanomas, which can explain the high susceptibility of BRAF V600E melanomas to bevacizumab treatment in a clinical phase III trial. WNT5A is a ligand of RTK ROR2 (receptor tyrosine kinase-like orphan receptor 2), which is upregulated by MAPK signaling in BRAF V600E cells (Figure 2b) [177]. In addition, the combination of the Wnt/βcatenin inhibitor pyrvinium with vemurafenib inhibited tumor growth in BRAF V600E HT-29 and COLO-205 CRC xenografts.…”