Li_2026_J.Mol.Neurosci_76_23

Ferroptosis, an iron-dependent regulated cell death form, is a key pathogenic mechanism in Alzheimer's disease (AD), especially in the entorhinal cortex, a brain region selectively vulnerable to early AD neuropathology. This study aimed to identify peroxiredoxin 6 (PRDX6) as a novel ferroptosis-related hub gene in the entorhinal cortex and validate its diagnostic and therapeutic potential in AD. Gene expression datasets (GSE138852, GSE5281, GSE48350, GSE118553) from the Gene Expression Omnibus (GEO) and ferroptosis-related genes (FRGs) from FerrDb were analyzed. Differential expressed genes (DEGs) were identified using Limma (|log2FC| > 1, P < 0.05), followed by Weighted Gene Coexpression Network Analysis (WGCNA) to delineate AD-associated modules. Machine learning approaches (LASSO and random forest) were employed to screen candidate hub genes, and CIBERSORT was utilized to assess correlations with immune cell infiltration. Single-cell RNA sequencing (scRNA-seq) data from GSE138852 mapped gene distribution across entorhinal cortex cell populations. Validation included analyses in the Alzdata database, receiver operating characteristic (ROC) curves for diagnostic accuracy, and Western blot assays in Abeta(1-42)-induced U251 astrocyte models. Functional enrichment analyses of WGCNA key module genes revealed involvement in anti-apoptosis regulation, cytosolic processes, enzyme binding, and the ferroptosis pathway. Machine learning identified six candidate genes, among which PRDX6 showed significant upregulation in the AD entorhinal cortex (Alzdata), correlation with both Abeta and tau pathologies, and a negative association with neutrophils. Single-cell profiling localized PRDX6 predominantly to astrocytes. ROC curves confirmed PRDX6 as the optimal hub gene, and Western blot validation demonstrated significantly elevated PRDX6 protein expression in Abeta(1-42)-induced U251 cells, consistent with bioinformatics findings. These findings establish PRDX6 as a pivotal mediator linking ferroptosis, immune cell dynamics, and AD neuropathology. Targeting PRDX6-mediated antioxidant pathways holds promise for intervening in ferroptosis-driven neurodegeneration and provides a novel avenue for AD diagnosis and therapeutic development.