Cross-dataset transcriptomics–proteomics analysis identifies Tlr2, Anxa5, and Grin2a as candidate mouse-brain signatures associated with chronic cerebral hypoperfusion

Abstract

Chronic cerebral hypoperfusion (CCH) is a neurological disorder characterized by persistent insufficiency of cerebral blood flow, yet specific biomarkers and mechanistic insights are lacking. This study integrated transcriptomic and proteomic multi-omics analyses, along with behavioral assessments and immunofluorescence validation in a bilateral common carotid artery stenosis (BCAS) mouse model, to identify and validate candidate multi-omics biomarkers of CCH and explore their potential druggability. Leveraging public transcriptomic datasets and in-house proteomic data, differential expression analysis, protein-protein interaction network construction, and five topological algorithms identified Tlr2 , Anxa5 , and Grin2a as candidate biomarkers. BCAS mice exhibited spatial memory impairments in behavioral tests. Immunofluorescence revealed upregulated TLR2 and ANXA5 , downregulated GRIN2A, and increased TLR2-IBA1 co-localization indicating microglial activation. Tlr2 and Anxa5 positively correlated with infiltrating immune cells, whereas Grin2a showed a negative correlation. A putative ceRNA network and in silico molecular docking suggested strong binding of pirinixic acid to both TLR2 and ANXA5. Tlr2 , Anxa5 , and Grin2a can serve as candidate multi-omics signature molecules of CCH in the mouse brain, with consistent alterations at the transcriptomic, proteomic, and tissue levels.