High miR-133a levels in the circulation anticipates presentation of clinical events in familial hypercholesterolaemia patients Rafael Escate 1,2† , Teresa Padro´ 1,2† , Rosa Suades 1 , Sandra Camino 1 , Ovidio Mu~ niz 3 , Jose Luis Diaz-Diaz 4 , Alessandro Sionis 5 , Pedro Mata 6 , and Lina Badimon 1,2,7 * 1 Cardiovascular-Program ICCC, Research Institute Hospital Santa Creu i Sant Pau, IIB-Sant Pau, Sant Antoni Maria Claret 167, 08025 Barcelona, Spain; 2 Centro de Investigacio´n Biome´dica en Red Cardiovascular (CIBERCV) Instituto de Salud Carlos III, Madrid, Spain; 3 Servicio de Medicina Interna, Hospital Virgen del Rocı ´o, Sevilla, Espa~na, Spain; 4 Department of Internal Medicine, Hospital Abente y Lago, A Coru~na, Spain; 5 Cardiology Department, Acute and Intensive Cardiac Care Unit, Hospital Santa Creu i Sant Pau, Barcelona, Spain; 6 Fundacio´n Hipercolesterolemia Familiar, Madrid, Spain; and 7 Cardiovascular Research Chair, UAB, Barcelona, Spain Received 11 May 2019; revised 17 November 2019; editorial decision 31 January 2020; accepted 10 February 2020; online publish-ahead-of-print 15 February 2020 Time for primary review: 29 days Aims Presentation of acute events in patients with atherosclerosis remains unpredictable even after controlling for classi- cal risk factors. MicroRNAs (miRNAs) measured in liquid biopsies could be good candidate biomarkers to improve risk prediction. Here, we hypothesized that miRNAs could predict atherosclerotic plaque progression and clinical event presentation in familial hypercholesterolaemia (FH) patients. .................................................................................................................................................................................................... Methods and results Circulating miRNAs (plasma, exosomes, and microvesicles) were investigated by TaqMan Array and RT-qPCR assays. Patients with genetic diagnosis of FH and healthy relatives from the SAFEHEART cohort were included. A differential signature of 10 miRNA was obtained by comparing two extreme phenotypes consisting of FH patients suffering a cardiovascular event (CVE) within a 8-year follow-up period (FH-CVE, N = 42) and non-FH hypercholes- terolaemic relatives from the same cohort, matched for age and treatment, without CVE during the same period (nFH-nCVE, N = 30). The validation studies included two independent groups of patients with FH background (dis- covery group, N = 89, validation group N = 196), developing a future CVE (FH-CVE) or not (FH-nCVE) within the same time period of follow-up. Of the 10 miRNAs initially selected, miR-133a was significantly higher in FH-CVE than in FH-nCVE patients. Receiver operating characteristic analysis confirmed miR-133a as the best microRNA for predicting CVE in FH patients (0.76 ± 0.054; P < 0.001). Furthermore, Kaplan–Meier and COX analysis showed that high plasma miR-133a levels associated to the higher risk of presenting a CVE within the next 8 years (hazard ratio 3.89, 95% confidence interval 1.88–8.07; P < 0.001). In silico analysis of curate biological interactions related miR- 133a with target genes involved in regulation of the cell-membrane lipid-receptor LRP6 and inflammatory cytokines (CXCL8, IL6, and TNF). These predictions were experimentally proven in human macrophages and endothelial cells transfected with agomiR-133a. .................................................................................................................................................................................................... Conclusion Elevated levels of miR-133a in the circulation anticipate those FH patients that are going to present a clinical CVE within the next 2 years (average). Mechanistically, miR-133a is directly related with lipid- and inflammatory signalling in key cells for atherosclerosis progression.                                                                                                                                                                                                                     * Corresponding author. Tel: þ34 93 5565882; fax: þ34 93 5565559, E-mail: lbadimon@santpau.cat † The first two authors contributed equally to the study. Published on behalf of the European Society of Cardiology. All rights reserved. VC The Author(s) 2020. For permissions, please email: journals.permissions@oup.com. Cardiovascular Research (2021) 117, 109–122 doi:10.1093/cvr/cvaa039 Downloaded from https://academic.oup.com/cardiovascres/article/117/1/109/5736549 by guest on 10 June 2022