Biomarker Profiling for Discrimination of High-Risk Asymptomatic Carotid Artery Stenosis Patients with Ulcerated Plaques: A Pilot Study

Although various methods are used to treat patients with asymptomatic carotid artery stenosis (ACAS), approaches are controversial, and combining imaging of carotid plaque features with biomarkers to identify plaques prone to rupture may be crucial in identifying high-risk ACAS patients. This study aimed to investigate a blood-based biomarker for discriminating ulceration in ACAS patients by analyzing plaque surface morphology through RNA sequencing of blood samples. Peripheral blood samples were collected from ACAS patients with plaque morphology determined by Doppler ultrasonography. Then, total RNA was isolated, and RNA-Seq was performed to analyze differentially expressed genes (DEGs). The KEGG, Reactome, and Gene Ontology (GO) terms pathway enrichment analyses were performed to investigate the molecular functions and biological processes involved in plaque formation. The pilot study included 7 ACAS patients, 57.1 % exhibiting ulcerated plaques. RNA-Seq results revealed significant upregulation of genes related to immune response, cell cycle regulation, and oxidative stress in ulcerated plaques. Especially, TP73, CCL3L3, and PXDNL genes showed the highest fold changes, indicating their role in endothelial damage, immune activation, and oxidative stress. KEGG and Reactome analyses identified TNF and chemokine signaling pathways as key regulators in ulcerated plaque formation. Our findings indicate that TP73, CCL3L3, and PXDNL may be potential biomarkers for identifying high-risk ACAS patients with ulcerated plaques due to their involvement in immune system regulation and oxidative stress-related processes. Thus, these genes and the pathways may be candidate biomarkers for early diagnosis and risk stratification, improving treatment approaches for ACAS.