Potential Ferroptosis-Related Genes and Immune Infiltration in Form-Deprivation Myopia: An Integrative Bioinformatics and in vitro Validation Study.

OBJECTIVE: This study aims to explore the molecular mechanisms of form-deprivation myopia (FDM) by identifying potential ferroptosis-related genes (FRGs) and characterizing the immune infiltration landscape, with the goal of providing preliminary insights that may inform future research on diagnostic or therapeutic candidates. METHODS: Transcriptomic data from GSE223434 (training set) and GSE210238 (validation set) were analyzed to identify differentially expressed FRGs (FRDEGs). Weighted gene co-expression network analysis (WGCNA) and machine learning algorithms, including LASSO regression and Random Forest, were employed to screen for hub genes. Immune infiltration was assessed using the CIBERSORT algorithm. Finally, the functional roles of hub genes were validated in vitro using TGF-β2-induced human scleral fibroblasts (HSFs) and siRNA-mediated knockdown experiments. RESULTS: A total of 256 FRDEGs were identified in FDM retinas. Three hub genes PRKAA1, TGFB3, and WWTR1, were selected and showed preliminary diagnostic potential value, with AUCs reaching 0.750, 0.725, and 0.762 in the training set. Immune analysis revealed significant differences in monocyte infiltration between myopia and control groups. In vitro assays confirmed that TGF-β2 treatment significantly elevated the expression of these hub genes and reduced GPX4 levels, while the ferroptosis inhibitor ferrostatin-1 (Fer-1) reversed these effects. Furthermore, silencing PRKAA1, WWTR1, or TGFB3 promoted ferroptosis and fibrotic markers, which were significantly rescued by Fer-1 intervention. CONCLUSION: The identified ferroptosis-immune axis, centered on PRKAA1, TGFB3, and WWTR1, may play a role in the pathogenesis of myopia. These hub genes represent candidate molecules that warrant further investigation as potential diagnostic markers, and they provide a preliminary biological perspective for understanding myopic scleral remodeling in the context of ferroptosis regulation.