Multi-omics and machine learning identify ALOX5 as a ferroptosis-associated driver of epithelial barrier dysfunction in chronic rhinosinusitis with nasal polyps.

Chronic rhinosinusitis with nasal polyps (CRSwNP) is a common chronic inflammatory disease of the upper airway, and its underlying pathogenesis remains incompletely understood. In this study, endoplasmic reticulum stress (ERS)-related gene sets were employed as the analytical starting point to systematically screen and identify core genes closely associated with CRSwNP. Through the integration of weighted gene co-expression network analysis (WGCNA), differential expression analysis, and multiple machine learning algorithms, 4 hub genes (ALOX5, HMOX1, PXDN, and CARD9) were identified, all of which exhibited favorable diagnostic performance for CRSwNP (AUC = 0.863-0.962, P < 0.05). RT-qPCR, Western blotting, and immunohistochemistry were performed to validate the aberrant expression of these hub genes in CRSwNP tissues. Notably, the expression level of ALOX5 was significantly positively correlated with the clinical severity of CRSwNP (R = 0.40-0.41, P < 0.01). Single-cell transcriptomic analysis further revealed that ferroptosis-related signatures were significantly enriched in nasal epithelial cells from patients with CRSwNP. Immunofluorescence staining and primary nasal epithelial cell experiments indicated that ALOX5 expression was negatively correlated with epithelial tight junction proteins and that targeting ALOX5 effectively modulated the ferroptosis process. Collectively, this study identified hub genes with potential diagnostic value and functional relevance in CRSwNP and elucidated the potential involvement of ALOX5-mediated epithelial ferroptosis in disease progression, thereby providing novel insights into the precise diagnosis and targeted therapy of CRSwNP.