Background

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by cognitive decline, memory loss and neuronal dysfunction. While amyloid-β plaques and tau tangles remain central to AD pathology, emerging evidence implicates ferroptosis, an iron-dependent, regulated form of cell death marked by lipid peroxidation and oxidative stress, as a critical contributor to disease progression.

Objective

This study investigates the interplay between major AD risk factors including chronic alcoholism, alcohol-nicotine co-abuse, aging, genetic predisposition, comorbidities and lifestyle habits and ferroptosis-related molecular pathways.

Methods

GEO datasets and 115 ferroptosis-related genes were analyzed using z-score/FDR and Limma, followed by GO/KEGG/GSEA enrichment, machine-learning–based gene selection with ROC-AUC validation, immune-cell profiling using CIBERSORT, and hub gene–miRNA network construction via NetworkAnalyst.

Results

Our findings demonstrate that these risk factors converge on shared mechanisms involving iron dysregulation, oxidative stress and lipid imbalance which are hallmarks of ferroptosis. We identified seven ferroptosis-associated biomarker genes CYBB, FERMT1, BAX, SOD1, ACSL4, TP53, and FTH1 as being significantly dysregulated in AD. Integrative gene-miRNA interaction analysis revealed several hub miRNAs including hsa-miR-34a (TP53, SOD1, BAX), hsa-miR-34b and hsa-miR-34c (TP53, FERMT1), hsa-miR-125a-5p (FERMT1, TP53) and hsa-miR-20a-5p (ACSL4, BAX) suggesting a network of coordinated post-transcriptional regulation. Additionally, we observed strong neuroinflammatory signatures in AD with increased infiltration of pro-inflammatory macrophages (M1), CD8⁺ T cells, monocytes and neutrophils. This heightened immune activity may be exacerbated by ferroptotic cell death and associated oxidative stress forming a vicious cycle of neurodegeneration. Finally, we propose glutathione and alpha-tocopheral (vitamin E) as potential therapeutic compounds due to their antioxidative role in preventing ferroptosis.

Conclusions

Overall, our study provides novel insights into the mechanistic connections between ferroptosis, miRNA regulation, immune responses and AD pathology highlighting potential biomarkers and therapeutic targets.