Abstract
The potential of extracellular circulating microRNAs (miRNAs) as non-invasive biomarkers of atrial fibrillation (AF) has been confirmed by a number of recent studies. However, the current data for some miRNAs are controversial and inconsistent, probably due to pre-analytical and methodological differences. In this work, we attempted to fulfill the basic pre-analytical requirements provided for circulating miRNA studies for application to paroxysmal atrial fibrillation (PAF) research. We used quantitative PCR (qPCR) to determine the relative plasma levels of circulating miRNAs expressed in the heart or associated with atrial remodeling or fibrillation with reported altered plasma/serum levels in AF: miR-146a-5p, miR-150-5p, miR-19a-3p, miR-21-5p, miR-29b-3p, miR-320a-3p, miR-328-3p, miR-375-3p, and miR-409-3p. First, in a cohort of 90 adult outpatient clinic patients, we found that the plasma level of miR-320a-3p was elevated in PAF patients compared to healthy controls and hypertensive patients without AF. We further analyzed the impact of medication therapies on miRNA relative levels and found elevated miR-320a-3p levels in patients receiving angiotensin-converting-enzyme inhibitors (ACEI) therapy. Additionally, we found that miR-320a-3p, miR-21-5p, and miR-146a-5p plasma levels positively correlated with the CHA2DS2-Vasc score and were elevated in subjects with CHA2DS2-Vasc ≥ 2. Our results indicate that, amongst the analyzed miRNAs, miR-320a-3p may be considered as a potential PAF circulating plasma biomarker, leading to speculation as to whether this miRNA is a marker of platelet state change due to ACEI therapy.
Highlights
New biomarkers of atrial fibrillation (AF), the most common arrhythmia with a high risk of morbidity and mortality, may improve the clinical risk assessment and understanding of the pathophysiology of this condition [1]
We first performed an analysis of plasma miRNAs in paroxysmal atrial fibrillation with a detailed evaluation of the main pre-analytical parameters required for the correct measurement of circulating miRNA biomarkers
Using hemolysis indices and the presence of concomitant diseases as confounding factors in statistical analysis, we observed a moderate increase in relative plasma levels of circulating hsa-miR-320a-3p in patients with paroxysmal atrial fibrillation (PAF) compared to healthy controls and hypertensive patients without AF
Summary
New biomarkers of atrial fibrillation (AF), the most common arrhythmia with a high risk of morbidity and mortality, may improve the clinical risk assessment and understanding of the pathophysiology of this condition [1]. The potential mechanism of changing the profile of extracellular circulating plasma miRNAs in AF is the secretion of exosomes by atrial cardiomyocytes. These exosomes contain a specific miRNA signature altered in AF [10]. Platelet elimination by centrifugation does not mean the elimination of PMPs, which are likely to be the main source of circulating miRNAs in plasma or serum [18,19] The levels of both platelet and serum miR-150 were lower in the presence of AF in patients with heart failure [20]. The analysis included miRNA species expressed in the heart or associated with atrial remodeling or fibrillation with reported plasma/serum levels altered in AF. We attempted to fulfill the basic pre-analytical requirements provided for circulating miRNA studies applied to PAF research
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