Differential dependency of human glioblastoma cells on vascular endothelial growth factor-A signaling via neuropilin-1

Abstract

Despite the high expression of neuropilin-1 (NRP-1) in human glioblastoma (GB), the understanding of its function as a co-receptor of vascular endothelial growth factor receptors (VEGFRs) in angiogenesis is currently limited. Therefore, the aim of the present study was to elucidate the non-classical function of NRP-1 expression in human GB. Expression patterns of NRP-1 and VEGF-A were determined by sandwich ELISA, western blot analysis, or immunohistochemistry. Differential dependency of GB cells following ablation of VEGF-A signaling was validated in vitro and in vivo. Cellular mechanism responsible for distinct response to VEGF-A signaling was evaluated by western blotting and immune-precipitation analysis. Prognostic implications were assessed using IHC analysis. GB cells exhibited differing sensitivity to silencing of vascular endothelial growth factor (VEGF)-A signaling, which resulted in a distinct expression pattern of wild-type or chondroitin-sulfated NRP-1. VEGF-A-sensitive GB exhibited the physical interaction between wild-type NRP-1 and FMS related receptor tyrosine kinase 1 (Flt-1) whereas VEGF-A-resistant GB exhibited chondroitin-sulfated NRP-1 without interaction with Flt-1. Eliminating the chondroitin sulfate modification in NRP-1 led to re-sensitization to VEGF-A signaling, and chondroitin sulfate modification was found to be associated with an adverse prognosis in patients with GB. The present study identified the distinct functions of NRP-1 in VEGF-A signaling in accordance with its unique expression type and interaction with Flt-1. The present research is expected to provide a strong basis for targeting VEGF-A signaling in patients with GB, with variable responses.