부산대학교 도서관

Funding Acknowledgements Type of funding sources: Public Institution(s). Main funding source(s): Agencia Estatal de Investigaciópn (AEI)" - Spanish Ministry of Economy Competitiveness of Science Agencia Estatal de Investigaciópn (AEI)" - Spanish Institute of Health Carlos III, ISCIII Background Familial hypercholesterolemia (FH) is associated with premature atherosclerosis but FH patients have a highly variable risk of clinical atherosclerotic cardiovascular disease (ACVD) presentation. Preclinical atherosclerosis has been identified by imaging of coronary artery calcium (CAC); however, little is known of the molecular pathways involved in vascular calcification in FH. Circulating microRNAs are proven as a powerful epigenetic regulators of atherosclerosis development. Purpose We have investigated the circulating microRNA signature of FH patients with CAC and asymptomatic atherosclerosis. Methods Differential plasma miRNA profiling was performed by Affymetrix microarray technology (discovery studies) and RT-PCR using TaqMan technology (validation studies) in plasma of FH subjects (N=86) included in a multicentric nationwide cohort. FH-patients with and without coronary calcification (determined by Agatston score) by computed tomographic angiography imaging were investigated. Proof-of-concept validation studies were performed in cell culture of human coronary-derived vascular smooth muscle cells (hcVSMC) calcification analysis. Results Using a non-targeted approach, a differential signature of 15 miRNA (all upregulated) was identified in FH-patients presenting subclinical atherosclerosis with calcified plaques (FH-CCS+, N=49, Agatston score >100) compared with FH-subjects without coronary calcification (FH-CCS-, N=37, Agatston score 0). After RT-PCR validation, 3 miRNAs (miR-193a-5p, miR-30e-5p and miR-6821-5p) were significantly upregulated in FH-CCS+ patients. ROC-analysis confirmed miR-6821-5p as the best miRNA for discriminating FH-patients with coronary calcification (AUC 0.70±0.06, P=0.003). High miR-6821-5p levels associated to older age (P=0.003) and high LDL burden (P=0.014), two established risk factors for calcification. In vitro studies showed miR-6821-5p up-regulation (P=0.018) in hcVSMC cultured under calcification conditions (high-phosphate medium) and expressing increased levels of calcification-related genes such as SPP1 (Secreted Phosphoprotein 1, osteopontin) and BMP2 (Bone Morphogenetic Protein 2) when compared with non-calcified hcVSMC. Conclusion Up-regulated levels of miR-6821-5p are found in FH patient with CAC+ atherosclerotic plaques, as well as in calcified isolated human coronary smooth muscle cells that express the pro-calcification genes as osteopontin and BMP2. miR-6821-5p is involved in the development of calcification in the coronary arteries and in the development of atherosclerotic plaques. [ABSTRACT FROM AUTHOR]