Differential miRNAs in acute spontaneous coronary artery dissection: Pathophysiological insights from a potential biomarker

Figures

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  Fig. 1. Plasma miRNAs differentially expressed in control subjects compared to AT-AMI patients. Box and whiskers Min to Max plots showing plasma levels of microRNAs. Data correspond to the expression of miRNAs in AT-AMI n=41 and control subjects n=9. Expression of miRNAs was analysed by RT-PCR using a serum/plasma focus PCR panel. Data were normalized using the global mean, expression levels are shown as Normalized relative quantity (NRQ), as indicated under Methods. Differences among groups were analysed by univariate test followed by FDR multiple correction test.

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  Fig. 2. Study design and cohorts. Plasma samples (AT-AMI n = 56, SCAD n = 26) were collected in both UK (AT-AMI n = 15, SCAD n = 15) and Spain (AT-AMI n = 41, SCAD n = 11), constituting the discovery and the validation cohorts, respectively. The number of males and females of each sample set is shown in the figure. Firstly, differences in the expression of plasma miRNAs between AT-AMI and SCAD patients from the discovery cohort were assessed performing RT-PCR assays using unbiased PCR panels of 179 miRNAs. Differentially expressed miRNAs identified in the discovery cohort were then analysed in 26 AT-AMI and 11 SCAD patients from the validation cohort using the same method. Lastly, resulting differentially expressed miRNAs were confirmed by performing PCR assays with individual PCR primer sets in 15 AT-AMI (all female) not included in the previous phase and 11 SCAD patients from the validation cohort. Created with BioRender.com

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  Fig. 3. Plasma microRNA signature and its predictive value to distinguish SCAD from AT-AMI patients. a, Box and whiskers Min to Max plots showing plasma levels of miR-let-7f-5p, miR-146a-5p, miR-151a-3p and miR-223-5p from SCAD (n = 11) and AT-AMI (n = 15) patients from validation cohort. Expression of miRNAs was analysed by RT-PCR using individual sets of primers. Data were normalized using the geometric mean of the five most stable miRNAs. Expression levels are shown as Normalized relative quantity (NRQ), as indicated under Methods. Differences among groups were analyzed by a Mann-Whitney test. b. Receiver operating characteristic (ROC) curves corresponding to miR-let-7f-5p, miR-146a-5p, miR-151a-3p and miR-223-5p expression levels from SCAD (n = 11) and AT-AMI (n = 15) patients from validation cohort. c. Receiver operating characteristic (ROC) curve corresponding to the concomitant evaluation of miR-let-7f-5p, miR-146a-5p, miR-151a-3p and miR-223-5p expression levels from SCAD (n = 11) and AT-AMI (n = 15) patients from validation cohort. b,c The bold line shows the ROC curve. The values inside the graph correspond to the area under the curve (AUC) and its 95% confidence interval; p…

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  Fig. 4. Target genes of miR-let-7f-5p, miR146a-5p, mir-151a-3p and miR-223-5p are associated with blood vessel biology. a, Bar graph illustrating the significance (-log p value) of blood vessel biology-associated biological processes predicted by PANTHER Classification System. Functional miRNA targets were selected using miRTarBase database and analysed to obtain functional classifications with terms from Gene Ontology. Data were analysed by Fisher's test and Bonferroni correction. b, Functional miRNA targets associated with blood vessel biology, TGF-beta pathway and cytoskeleton are shown. Colours indicate each miRNA with its corresponding targets. SMC, smooth muscle cell. Triangles indicate cytokines and soluble factors; squares indicate plasma membrane receptor proteins; circles, intracellular proteins (cytoplasmic and/or nuclear); hexagons, proteins with extra- and intracellular function and/or presence. c, Box and whiskers Min to Max plots showing plasma levels of CXCL8, TGFB1, TGFBR1, Endothelin-1 and MMP2 in SCAD patients (n = 26) and AT-AMI patients (n = 34). Levels of these molecules were analysed using a custom multiplex ELISA assay. Differences between groups were evaluated…