Ferroptosis-associated candidate genes in aneurysmal subarachnoid hemorrhage: evidence from human peripheral blood and mouse brain.

Abstract

Background: Aneurysmal subarachnoid hemorrhage (aSAH) has high morbidity and mortality. Ferroptosis may contribute to aSAH-related brain injury, but its peripheral and brain-associated signatures remain unclear.

Methods: Human peripheral-blood dataset GSE36791 (43 aSAH, 18 controls) was analyzed using limma, FerrDb intersection, WGCNA, GO/KEGG enrichment, and PPI-based prioritization. qPCR validation was performed in peripheral blood from 10 aSAH patients and 10 non-SAH controls; aSAH samples were collected within 24 h after rupture. Mean ages were 60.2 ± 7.7 and 63.6 ± 8.9 years, with 5 and 4 males, respectively. Brain-level validation used GSE79416 mouse cortex and OMIX006611 mouse SAH single-cell RNA-seq data. Male C57BL/6 J mice (8 weeks, 20-22 g) underwent cisterna magna autologous blood injection; brain tissues were collected 1 day after SAH (n = 6/group). Immune infiltration and molecular docking were also performed.

Results: Among 2,645 DEGs, 35 were ferroptosis-related. MAPK3, STAT3, G6PD, and PGD were prioritized as ferroptosis-associated candidate genes and were upregulated in human aSAH peripheral blood, mouse SAH cortex, and the clinical qPCR cohort. Single-cell analysis showed expression of the corresponding mouse genes across annotated brain cell types, especially immune/glial populations. In mouse SAH brain tissue, MAPK3/ERK1, STAT3, PGD, and G6PD protein levels increased, p-STAT3/STAT3 ratio was elevated, MDA content increased, and GPX4 activity decreased. Immune profiling showed increased neutrophils, mast cells, and activated dendritic cells. Ganoderic acid A was prioritized as a preliminary in silico candidate.

Conclusion: MAPK3, STAT3, G6PD, and PGD are ferroptosis-associated candidate genes in aSAH, supported by human peripheral-blood, mouse cortex, single-cell, and mouse brain validation. Ganoderic acid A remains an exploratory in silico candidate requiring further functional validation.