Circulating MicroRNA Profiling in Non-ST Elevated Coronary Artery Syndrome Highlights Genomic Associations with Serial Platelet Reactivity Measurements

Abstract

changes in platelet physiology are associated with simultaneous changes in microRnA concentrations, suggesting a role for microRnA in platelet regulation. Here we investigated potential associations between microRnA and platelet reactivity (pR), a marker of platelet function, in two cohorts following a non-St elevation acute coronary syndrome (nSte-AcS) event. first, non-targeted microRnA concentrations and pR were compared in a case (n = 77) control (N = 76) cohort within the larger TRILOGY-ACS trial. MicroRNA significant in this analysis plus CVD-associated microRNAs from the literature were then quantified by targeted rt-PCR in the complete TRILOGY-ACS cohort (N = 878) and compared with matched PR samples. Finally, microRNA significant in the non-targeted & targeted analyses were verified in an independent post NSTE-ACS cohort (N = 96). From the non-targeted analysis, 14 microRNAs were associated with PR (Fold Change: 0.91-1.27, p-value: 0.004-0.05). From the targeted analysis, five microRNAs were associated with PR (Beta: −0.09-0.22, p-value: 0.004-0.05). Of the 19 significant microRNAs, three, miR-15b-5p, miR-93 and miR-126, were consistently associated with pR in the tRiLoGY-AcS and independent Singapore post-AcS cohorts, suggesting the measurement of circulating microRnA concentrations may report on dynamic changes in platelet biology following a cardiovascular ischemic event.Despite advancements in its management, cardiovascular disease including non-ST elevation acute coronary syndrome (NSTE-ACS) remains a major cause of patient morbidity and mortality 1-4 . Antiplatelet therapies including P2Y 12 antagonists are used to treat NSTE-ACS acutely and in the longer-term. While measurements of platelet reactivity (PR) have facilitated the rapid quantification of anti-platelet medication efficacy, questions still remain about the utility of PR to predict clinical outcomes 5-8 . Platelet genetics and PR sub-studies from large-scale clinical trials have informed our understanding of individual variability in P2Y 12 inhibitor response, but genomic regulation of individual patient's PR has not been fully clarified 9-14 .Small non-coding ribonucleic acids (RNA) including microRNAs have emerged as potential regulators of PR in patients with cardiovascular disease 15 . MicroRNAs (miRNAs) are 18-22 nucleotide non-coding RNAs that play an essential role in gene modulation and expression and are established regulators of cardiovascular

MicroRNA as Biomarkers for Platelet Function and Maturity in Patients with Cardiovascular Disease

“…[33][34][35][36][37][38][39][40][41][42][43][44][45][46] Among the studies using plasma samples, seven studies used platelet-rich plasma (PRP), 34,36,37,39,42,43,46 whereas two studies used plateletpoor plasma (PPP). 33,35 To compensate for potential analytical variations, 47 all studies used normalization strategies of the measured miRs. Four studies used high-throughput sequencing to investigate a large number of miRs followed by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) of a selected number of miRs.…”

“…31,32,[34][35][36][37][38][40][41][42][43]45 Subgroups, Laboratory Analyses, and Treatment In 12 of 16 included studies, the patient cohort was divided into subgroups. [32][33][34][35][36][37]39,[42][43][44][45][46] The methods and cut-offs employed to define subgroups varied between studies.…”

“…31 In five of the included studies, blood samples were obtained at different time points. 33,34,37,40,43 The results were only presented if the studies also investigated the corresponding association between miRNA and platelet function. We denominated the first blood sample as baseline and the following blood samples were named after the number of days they were taken after the baseline sample.…”

“…In studies on patients with acute coronary syndrome (ACS), all patients were treated according to guidelines with dual antiplatelet inhibitors consisting of aspirin and clopidogrel preceded by loading dose (►Table 2). 33,34,38,39,42,43,45 In the studies on patients with coronary artery disease (CAD), the antiplatelet treatment consisted of either aspirin monotherapy or dual antiplatelet therapy with aspirin plus adenosine diphosphate (ADP) inhibitor (►Table 3). 32,33,36,37,40,41,44 In the two studies examining patients with intermittent claudication 35 or patients with symptomatic atherothrombotic diseases, 46 all patients were treated with aspirin monotherapy (►Table 4).…”

MicroRNA as Potential Biomarkers of Platelet Function on Antiplatelet Therapy: A Review

“…Furthermore, these miRNAs were upregulated in activated platelet supernatant, which suggests the role of platelet reactivity for their release, although the phenomenon was not reflected in peripheral circulation (Gidlöf et al, 2013). In the case of NSTEMI patients, correlation between miR-15b-5p, miR-93, and miR-126 expression levels and platelet reactivity was observed (Becker et al, 2020).…”