Experimental validation and identification of ferroptosis-associated biomarkers for diagnostic and therapeutic targeting in hearing loss.

OBJECTIVES: Ferroptosis, a regulated form of cell death, has attracted significant attention in hearing loss research; however, the role of ferroptosis-related genes remains unclear. This study aimed to clarify diagnostic and therapeutic targeting of ferroptosis-related genes in hearing loss. METHODS: Differentially expressed genes related to hearing loss from the GEO database were intersected with ferroptosis-related genes. The Lasso and SVM-RFE models were applied to reduce the gene set, identifying model genes. Biological functions, pathways, and gene-drug associations related to these model genes were analyzed. Age-related hearing loss (ARHL) genes within the model genes were obtained from a genome-wide association study (GWAS) dataset. Further validation was conducted in HEI-OC1 cells and the cochleae of C57BL/6J mice, including auditory brainstem response (ABR) testing, qRT-PCR, Western blotting, Fe(2+) detection, and immunofluorescence analysis. RESULTS: The study identified 20 ferroptosis-related genes associated with hearing loss. Using Lasso and SVM-RFE models, a novel model was constructed, consisting of nine genes (SCD, ENPP2, PANX2, NEDD4, MEF2C, ABCC5, KLHDC3, CYP4F8 and IFNA2). Among these, MEF2C and NEDD4 were found to be associated with ARHL. CONCLUSION: Ferroptosis is a potential pathological mechanism in hearing loss research, and the nine ferroptosis-related genes identified provide promising targets for exploring new diagnostics and treatments for hearing loss. Notably, MEF2C and NEDD4 are associated with ARHL.