Ferroptosis regulator NOX1 acts a diagnostic biomarker and mediates disease progression with the transcriptional regulation of STAT3 in glaucoma

PURPOSE: Primary open-angle glaucoma (POAG) is the leading cause of irreversible blindness. Regrettably, the roles of ferroptosis-related (FR) genes in POAG remain elusive. DESIGN: Five GEO data sets and a series of experimentations in vitro were used for bioinformatic exploration and biological validation. METHODS: Using multiple machine learning algorithms, four critical FR genes in POAG progression were screened. The clinical value and biological function of NOX1 were comprehensively analyzed using bioinformatic approaches. A POAG in vitro model was constructed using H2O2 treatment. NOX1 effects on the viability of retinal ganglion cells (RGCs) and ferroptosis process were determined through CCK8, EdU, ROS detection, and transmission electron microscopy. Its upstream transcriptional mechanisms were determined through dual luciferase assays, and chromatin immunoprecipitation (ChIP). RESULTS: NOX1 was identified as the critical FR gene in POAG progression and served as an effective diagnostic biomarker. High-NOX1 expression was tightly associated with increased infiltration levels of multiple subtypes of T cells, such as T cells CD8 and T cells CD4. However, the enrichments of eight metabolic gene sets did not differ between the POAG samples with high- and low-NOX1 expression groups. Silencing NOX1 maintained RGC survival and inhibited the ferroptosis process. Mechanistically, STAT3 upregulated NOX1 by binding its promoter region that was located at the 429 th to 419 th bases upstream of the NOX1 transcriptional start site. NOX1 overexpression reversed the inhibitory effects of silencing STAT3 on RGC survival and the ferroptosis process. CONCLUSIONS: NOX1 was a good biomarker for characterizing POAG and promoted POAG progression through STAT3-mediated transcriptionally activation.