High miR-126-3p levels associated with cardiovascular events in a general population

Figures

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  Fig. 1. Study design for miRNA identification associated with albuminuria and cardiovascular events. The study design involved a discovery phase including hypertensives (HTN) (n = 48) with urinary albumin excretion (UAE) or normoalbuminuric (Non-UAE) by small RNA sequencing (Small RNA-Seq). In a second validation stage with a large hypertensive cohort (n = 69), including UAE or Non-UAE, and healthy controls (CNT, n = 29), 5 microRNA (miRNAs) were validated by real-time quantitative polymerase chain reaction (RT-qPCR). Finally, in a testing population cohort (Hortega's cohort), at baseline (n = 1025), the associations with 3 miRNAs (miR-374a-5p, miR-1260b and miR-126–3p) and albuminuria were tested. Then, at the 14-year follow-up (n = 926), the predictive power of miRNA levels with cardiovascular disease (CVD) events and combined coronary artery disease (CAD) + stroke incidence was investigated.

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  Fig. 2. Discriminatory power of plasma miRNAs for urinary albumin excretion. (a) Plasma levels of miR-374a-5p, miR-1260b and miR-126–3p in testing cohort 1025 participants, 83 with urinary albumin excretion (UAE). The microRNA (miRNA) levels have been quantified by real-time quantitative polymerase chain reaction (RT-qPCR), expressed as log10-transformed of absolute copy number and was normalized to miR-125a and miR-186 as internal controls. The miR-374a-5p, miR-1260b and miR-126–3p were significantly increased in UAE subjects. Data was compared using the Mann-Whitney U test. The ends of the boxes indicate the quartile 1(upper) and 3 (down), the middle line represents the median, and the whiskers are the minimum and maximum values of the data series (b) The areas under the curve (AUC) and receiver operating characteristics (ROC) curves were calculated for plasma miRNAs in the testing cohort. The diagnostic signature was calculated by a logistic regression model. The AUC of 3-miRNAs and 2-miRNAs signatures were significantly higher when compared to individual miRNAs (p < 0.05). (c) ROC curves are shown for individual and miRNA signatures.

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  Fig. 3. Kaplan–Meier of cardiovascular event-free survival. Testing cohort was grouped according to plasma miR-126–3p, with the value -0.92 as the cut-off obtained from receiver operating characteristic (ROC) analysis during the follow-up period of 14-years for cardiovascular disease (CVD) incidence (a) and for combined coronary artery disease (CAD) and stroke (b). Higher levels of miR-126–3p were associated with a significantly lower incidence rate of CVD and combined CAD and stroke. Weighted long-rank test: p-values at bottom survival curves are shown.