Inhibition of fibroblast growth factor receptor with AZD4547 mitigates juvenile nasopharyngeal angiofibroma

Abstract

Background Juvenile nasopharyngeal angiofibroma (JNA) is a benign tumor that presents in adolescent males. Although surgical excision is the mainstay of treatment, recurrences complicate treatment. There is a need to develop less invasive approaches for management. JNA tumors are composed of fibroblasts and vascular endothelial cells. We identified fibroblast growth factor receptor (FGFR) and vascular endothelial growth factor (VEGF) expression in JNA-derived fibroblasts. FGFR influences fibroblast proliferation and VEGF is necessary for angiogenesis. We hypothesized that targeting FGFR would mitigate JNA fibroblast proliferation, invasion, and migration, and that targeting the VEGF receptor would attenuate endothelial tubule formation. Methods After informed consent, fibroblasts from JNA explants of 3 patients were isolated. Fibroblasts were treated with FGFR inhibitor AZD4547, 0 to 25 µg/mL for 72 hours and proliferation was quantified using CyQuant assay. Migration and invasion of JNA were assessed using 24-hour transwell assays with subsequent fixation and quantification. Mitigation of FGFR and downstream signaling was evaluated by immunoblotting. Tubule formation was assessed in human umbilical vein endothelial cells (HUVECs) treated with vehicle control (dimethylsulfoxide [DMSO]) or semaxanib (SU5416) as well as in serum-free media (SFM) or JNA conditioned media (CM). Tubule length was compared between treatment groups. Results Compared to control, AZD4547 inhibited JNA fibroblast proliferation, migration, and invasion through inhibition of FGFR and downstream signaling, specifically phosphorylation of - p44/42 mitogen activated protein kinase (p44/42 MAPK). JNA fibroblast CM significantly increased HUVEC tubule formation (p = 0.0039). Conclusion AZD4547 effectively mitigates FGFR signaling and decreases JNA fibroblast proliferation, migration, and invasion. SU5416 attenuated JNA fibroblast-induced tubule formation. AZD4547 may have therapeutic potential in the treatment of JNA.

Differential Gene Expression and Pathway Analysis in Juvenile Nasopharyngeal Angiofibroma Using RNA Sequencing

“…This builds upon previous understanding that basic fibroblast growth factor (bFGF), a pleiotropic growth factor acting as a mitogen for both fibroblast and endothelial cells, is strongly associated with disease progression. 13,14 Thus, our differential expression analysis corroborates current understanding of drivers underlying this disease while also detailing novel pathways not previously associated.…”

“…This is the first evidence of FGFR overexpression in JNA patient specimens and supports targeted therapy against FGFR in JNA patients. 14…”

Early Postoperative Magnetic Resonance in the Diagnosis of Persistent Juvenile Angiofibroma

“…In extreme scenarios (i.e., growing residues in critical areas), non-surgical approaches, such as radiotherapy or medical treatments (antiandrogens, corticosteroids, beta-blockers, and antiVEGF), can be considered. [2][3][4][5][26][27][28][29][30][31][32][33][34][35] The current role of radiotherapy is limited to lesions involving critical intracranial structures not amenable to surgery despite the high control rates reported because of morbidity concerns. 15,26,27 The main issue limiting its use in JA is the young age of patients and the poorly known long-term effects, even if usually administered with low radiation doses and with a technique like intensity-modulated radiotherapy or radiosurgery.…”

Inhibition of CK2 Reduces NG2 Expression in Juvenile Angiofibroma

“…In the last years, intensive research efforts have been made to find alternative strategies for the treatment of JA [5][6][7][8]. Targeting specific proteins that control JA growth and invasion may represent a particularly effective therapeutic approach [7,9].…”