High-density lipoprotein remodelled in hypercholesterolaemic blood induce epigenetically driven down-regulation of endothelial HIF-1 alpha expression in a preclinical animal model
Por:
Ben-Aicha, S, Escate, R, Casani, L, Padro, T, Pena, E, Arderiu, G, Mendieta, G, Badimon, L, Vilahur, G
Publicada:
1 jun 2020
Resumen:
Aims High-density lipoproteins (HDLs) are circulating micelles that transport proteins, lipids, and miRNAs. HDL-transported miRNAs (HDL-miRNAs) have lately received attention but their effects on vascular cells are not fully understood. Additionally, whether cardiovascular risk factors affect HDL-miRNAs levels and miRNA transfer to recipient cells remains equally poorly known. Here, we have investigated the changes induced by hypercholesterolaemia on HDL-miRNA levels and its effect on recipient endothelial cells (ECs).
Methods and results Pigs were kept on a high-fat diet (HC; n = 10) or a normocholesterolaemic chow (NC; n = 10) for 10 days reaching cholesterol levels of 321.0 (229.7-378.5) mg/dL and 74.0 (62.5-80.2) mg/dL, respectively. HDL particles were isolated, purified, and quantified. HDL-miRNA profiling (n = 149 miRNAs) of HC- and NC-HDLs was performed by multipanel qPCR. Cell cultures of porcine aortic ECs were used to determine whether HDL-miRNAs were delivered to ECs. Potential target genes modulated by miRNAs were identified by bioinformatics and candidate miRNAs were validated by molecular analysis. In vivo effects in the coronary arteries of normocholesterolaemic swine administered HC- or NC-HDLs were analysed. Among the HDL-miRNAs, four were found in different amounts in HC- and NC-HDL (P < 0.05). miR-126-5p and -3p and miR-30b-5p (2.7x, 1.7x, and 1.3x, respectively) were found in higher levels and miR-103a-3p and miR-let-7g-5p (-1.6x, -1.4x, respectively) in lower levels in HC-HDL. miR-1265-p and -3p were transferred from HC-HDL to EC (2.5x; P < 0.05), but not from NC-HDL, by a SRB1-mediated mechanism. Bioinformatics revealed that HIF1 alpha was the miR-126 target gene with the highest predictive value, which was accordingly found to be markedly reduced in HC-HDL-treated ECs and in miR126 mimic transfected ECs. In vivo validation confirmed that HIF1 alpha was diminished in the coronary endothelial layer of NC pigs administered HC-HDL vs. those administered NC-HDL (P < 0.05).
Conclusion Hypercholesterolaemia induces changes in the miRNA content of HDL enhancing miR126 and its delivery to ECs with the consequent down-regulation of its target gene HIF1 alpha.
Filiaciones:
Ben-Aicha, S:
Hosp Santa Creu & Sant Pau, IIB St Pau, Cardiovasc Program ICCC, Res Inst, Barcelona, Spain
Univ Barcelona UB, Sch Med, Barcelona, Spain
Escate, R:
Hosp Santa Creu & Sant Pau, IIB St Pau, Cardiovasc Program ICCC, Res Inst, Barcelona, Spain
Inst Salud Carlos III, Ctr Invest Biomed Red Cardiovasc CIBERCV, Madrid, Spain
Casani, L:
Hosp Santa Creu & Sant Pau, IIB St Pau, Cardiovasc Program ICCC, Res Inst, Barcelona, Spain
Padro, T:
Hosp Santa Creu & Sant Pau, IIB St Pau, Cardiovasc Program ICCC, Res Inst, Barcelona, Spain
Inst Salud Carlos III, Ctr Invest Biomed Red Cardiovasc CIBERCV, Madrid, Spain
Pena, E:
Hosp Santa Creu & Sant Pau, IIB St Pau, Cardiovasc Program ICCC, Res Inst, Barcelona, Spain
Inst Salud Carlos III, Ctr Invest Biomed Red Cardiovasc CIBERCV, Madrid, Spain
Arderiu, G:
Hosp Santa Creu & Sant Pau, IIB St Pau, Cardiovasc Program ICCC, Res Inst, Barcelona, Spain
Mendieta, G:
Hosp Santa Creu & Sant Pau, IIB St Pau, Cardiovasc Program ICCC, Res Inst, Barcelona, Spain
Univ Barcelona UB, Sch Med, Barcelona, Spain
Hosp Clin Barcelona, Cardiol Dept, Barcelona, Spain
Badimon, L:
Hosp Santa Creu & Sant Pau, IIB St Pau, Cardiovasc Program ICCC, Res Inst, Barcelona, Spain
Inst Salud Carlos III, Ctr Invest Biomed Red Cardiovasc CIBERCV, Madrid, Spain
Univ Autonoma Barcelona UAB, Cardiovasc Res Chair, Barcelona, Spain
Vilahur, G:
Hosp Santa Creu & Sant Pau, IIB St Pau, Cardiovasc Program ICCC, Res Inst, Barcelona, Spain
Inst Salud Carlos III, Ctr Invest Biomed Red Cardiovasc CIBERCV, Madrid, Spain
Green Published, hybrid
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