Patient-derived 3D nasal spheroids reveal epithelial changes following Dupilumab therapy in CRSwNP: a preliminary report

Chronic rhinosinusitis with nasal polyps (CRSwNP) is a type 2 inflammatory disease associated with epithelial dysfunction and impaired mucosal barrier integrity. Dupilumab, an IL-4 receptor alpha antagonist, has shown clinical efficacy, but its cellular effects on nasal epithelium remain poorly understood. Advanced in vitro models such as 3D spheroid cultures may provide insight into epithelial organization under treatment. We conducted a preliminary study using nasal epithelial cells obtained from three patient groups: CRSwNP treated with Dupilumab for 16 weeks (n = 3), untreated CRSwNP (n = 3), and turbinate hypertrophy controls (n = 3). Cells were isolated by enzymatic digestion and cultured in ultra-low attachment plates using sphere-promoting medium to assess spheroid formation. Observations were performed using phase-contrast microscopy. Due to the limited sample size, data were analyzed qualitatively without statistical testing. Control cells formed compact spheroids, while untreated CRSwNP cells failed to generate structured spheroids, showing only aggregates. Cells from Dupilumab-treated patients produced well-organized spheroids, suggesting improved epithelial organization. Occasional surface movement was observed but not quantitatively assessed. No molecular or ultrastructural assays were performed to confirm mechanistic hypotheses. Our preliminary findings indicate that Dupilumab treatment may be associated with improved epithelial organization in CRSwNP, as shown by spheroid formation in 3D culture. However, these observations are preliminary and based on a small cross-sectional cohort. Future studies should include longitudinal sampling, functional assays, and molecular analyses to confirm mechanisms and validate these results.