Circulating Exosomal miRNAs as Novel Biomarkers for Stable Coronary Artery Disease

Abstract

Exosomal miRNAs are currently being explored as a novel class of biomarkers in cardiovascular diseases. However, few reports have focused on the value of circulating exosomal miRNAs as biomarkers for stable coronary artery disease (SCAD). Here, we aimed to investigate whether miRNAs involved in cardiovascular diseases in circulating exosomes could serve as novel diagnostic biomarkers for SCAD. Firstly, the serum exosomes were isolated and purified by the ExoQuick reagent and identified by transmission electron microscopy, western blot, and nanoparticle tracking analysis. Then, the purified exosomes were quantified by measuring the exosome protein concentration and calculating the total protein amount. Next, eight miRNAs involved in cardiovascular diseases, miR-192-5p, miR-148b-3p, miR-125a-3p, miR-942-5p, miR-149-5p, miR-32-5p, miR-144-3p, and miR-142-5p, were quantified in circulating exosomes from the control group ( n = 20 ) and the SCAD group ( n = 20 ) by quantitative real-time polymerase chain reaction (qPCR). Finally, the gene targets of the differentially expressed miRNAs were predicted, and the functions and signaling pathways of these targets were analyzed using an online database. The isolated exosomes had a bilayer membrane with a diameter of about 100 nm and expressed exosomal markers including CD63, Tsg101, and Flotillin but negatively expressed Calnexin. Both the exosome protein concentration and total protein amount exhibited no significant differences between the two groups. The qPCR assay demonstrated that among the eight miRNAs, the expression levels of miR-942-5p, miR-149-5p, and miR-32-5p in the serum exosomes from the SCAD group were significantly higher than that from the control group. And the three miRNAs for SCAD diagnosis exhibited AUC values of 0.693, 0.702, and 0.691, respectively. GO categories and signaling pathways analysis showed that some of the predictive targets of these miRNAs were involved in the pathophysiology processes of SCAD. In conclusion, our findings suggest that serum exosomal miR-942-5p, miR-149-5p, and miR-32-5p may serve as potential diagnostic biomarkers for SCAD.

Magic and mystery of microRNA‐32

“…The diagnosis AUC value is 0.691, which suggest that serum exosomal miR-32-5p may serve as potential diagnostic biomarkers for SCAD. 24 Similar results were reported from another study, patient with CAD (All CAD patients were confirmed by angiographic evidence of >70% stenosis of at least 1 main coronary artery), plasma miR-32-3p was significantly higher than the control group, and the diagnosis AUC value is 0.745 (95%CI 0.649-0.84). 25 All indicate that miR-32 has promising diagnostic value for CAD.…”

“…The expression levels of miR‐32‐5p were significantly higher in circulating exosomes from patient with stable coronary artery disease (SCAD) than those from the control group. The diagnosis AUC value is 0.691, which suggest that serum exosomal miR‐32‐5p may serve as potential diagnostic biomarkers for SCAD 24 . Similar results were reported from another study, patient with CAD (All CAD patients were confirmed by angiographic evidence of >70% stenosis of at least 1 main coronary artery), plasma miR‐32‐3p was significantly higher than the control group, and the diagnosis AUC value is 0.745 (95%CI 0.649–0.84) 25 .…”

Association between Exosomal miRNAs and Coronary Artery Disease by Next-Generation Sequencing

“…The expression of exosomal miRNA-146a was significantly up-regulated in patients with heart failure, and that association was not found from the circulating plasma miRNA-146a. Therefore, exosomal miRNAs have greater potential for investigating cardiovascular diseases than circulating plasma miRNAs [ 19 ]. To the best of our knowledge, the current study is the largest study to explore the varied expression of exosomal miRNAs in stable three-vessel CAD by NGS.…”

“…Our study possesses several strengths as compared to prior studies. In the study conducted by Zhang et al, eight miRNAs known to be involved in the pathogenesis of cardiovascular diseases (miR-192-5p, miR-148b-3p, miR-125a-3p, miR-942-5p, miR-149-5p, miR-32-5p, miR-144- 3p, and miR-142-5p) were selected to investigate the association between exosomal miRNAs and CAD [ 19 ]. Since only the eight candidate miRNAs were examined, other previously unidentified exosomal miRNAs associated with CAD were likely missed in their study.…”

Engineering of Extracellular Vesicles Based on Payload Changes for Tissue Regeneration

“…At this time, the circRNA profile was significantly changed in the bone marrow-derived exosomes of ET patients, of which cir-cDAP3, circASXL1, and circRUNX1 tended to decrease in ET patients. Thus, this study provides insights into the role of circRNA in ET pathogenesis and reported that circRNA, as well as miRNAs involved in cellular communication, will provide a predictable basis for predictive diagnosis and treatment of their potential functions [102].…”