Plasma NOTCH3 and the risk of cardiovascular recurrence in patients with ischemic stroke

Abstract

Background: Ischemic stroke patients are more prone to developing another cardiovascular event. Aim: This study aims to examine potential biological predispositions to cardiovascular recurrence in patients with ischemic stroke. Design: Human and preclinical studies. Methods: Quantitative proteomic analysis, animal stroke, atherosclerosis models and circulating endothelial cells (CECs) were employed to examine candidate biomarkers derived from an ischemic stroke cohort in Singapore. Results Proteomic analysis of pooled microvesicles of 'Event' (n = 24) and without 'Event' (n = 24) samples identified NOTCH3 as a candidate marker; plasma NOTCH3 were shown to be elevated in 'Event' patients compared to those without 'Events' and age-matched controls. In a validation cohort comprising 431 prospectively recruited ischemic stroke patients (mean age 59.1 years; median follow-up 3.5 years), men with plasma NOTCH3 (>1600 pg/ml) harbored increased risk of cardiovascular recurrence (adjusted hazards ratio 2.29, 95% CI 1.10-4.77); no significant association was observed in women. Chronic renal failure, peripheral artery disease and NT-pro-brain natriuretic peptide were significant predictors of plasma NOTCH3 in men without ischemic stroke (adjusted r(2 )= 0.43). Following middle cerebral artery occlusion, NOTCH3 expression in mouse sera increased and peaked at 24 h, persisting thereafter for at least 72 h. In Apoe(-/-) atherosclerotic mice, NOTCH3 stained the endothelium of defective arterial lining and atherosclerotic plaques. Analysis of CECs isolated from stroke patients revealed increased gene expression of NOTCH3, further supporting endothelial damage underpinning NOTCH3-mediated atherosclerosis. Conclusion: Findings from this study suggests that NOTCH3 could be important in cardiovascular recurrence following an ischemic stroke. Graphical Abstract