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Seven Encapsulated and Nonencapsulated (iso)microRNAs in Human Samples Predict Coronary Atherosclerosis and Cardiac Events

Coronary Artery Disease (CAD) is a chronic inflammatory process that modifies microRNA (miR) abundance, intracellularly, in vascular cells, and extracellularly, in plasma. Extracellular miRs are encapsulated in microvesicles (MV) or protein‐bound, and variations in miR profiles are promising as novel biomarkers of CAD severity. We hypothesized that CAD‐associated variations in the miR profile are related to changes in the abundance of miR isoforms (isomiRs). IsomiR sequences differ by 1‐3 nucleotides from the consensus miR sequences. Using RNA‐deep sequencing, the change in abundance/frequency of isomiRs was assessed in human blood samples from cohorts with varying CAD severity. This analysis highly expressed 18 miRs whose dominant form was an isomiR of a consensus miR. Using custom‐designed RT/PCR primers for the 18 isomiRs, unfractionated and fractionated plasma from patients with/without significant CAD were assessed for isomiR concentrations. Seven isomiRs (miR‐10b, 30d, 93, 143, 181a, 182, and 744) were detectable and distinctly different in abundance between cohorts. Subgroupings of these 7 isomiRs (based on abundance/frequency within each plasma fraction) were examined to develop an isomiR biomarker panel. Statistical comparison and correlation analysis of this subgrouping had an accuracy of 69‐100% in predicting relevant clinical outcomes such as CAD severity, MV counts (inflammation marker), or myocardial infarction. These data support the development and clinical use of an isomiR biomarker panel to assess CAD risk.