de Gonzalo-Calvo David, Cenarro Ana, Garlaschelli Katia, Pellegatta Fabio, Vilades David, Nasarre Laura, Camino-Lopez Sandra, Crespo Javier, Carreras Francesc, Leta Rubén, Catapano Alberico Luigi, Norata Giuseppe Danilo, Civeira Fernando, Llorente-Cortes Vicenta
Group of Lipids and Cardiovascular Pathology, Biomedical Research Institute Sant Pau (IIB Sant Pau), Barcelona, Spain; CIBERCV, Instituto de Salud Carlos III, Madrid, Spain.
CIBERCV, Instituto de Salud Carlos III, Madrid, Spain; Lipid Unit and Molecular Research Laboratory, IIS Aragón, Hospital Universitario Miguel Servet, Universidad de Zaragoza, Zaragoza, Spain.
J Mol Cell Cardiol. 2017 May;106:55-67. doi: 10.1016/j.yjmcc.2017.03.005. Epub 2017 Mar 23.
To analyze the impact of atherogenic lipoproteins on the miRNA signature of microvesicles derived from human coronary artery smooth muscle cells (CASMC) and to translate these results to familial hypercholesterolemia (FH) and coronary artery disease (CAD) patients.
Conditioned media was collected after exposure of CASMC to atherogenic lipoproteins. Plasma samples were collected from two independent populations of diagnosed FH patients and matched normocholesterolemic controls (Study population 1, N=50; Study population 2, N=24) and a population of patients with suspected CAD (Study population 3, N=50). Extracellular vesicles were isolated and characterized using standard techniques. A panel of 30 miRNAs related to vascular smooth muscle cell (VSMC) (patho-)physiology was analyzed using RT-qPCR.
Atherogenic lipoproteins significantly reduced levels of miR-15b-5p, -24-3p, -29b-3p, -130a-3p, -143-3p, -146a-3p, -222-3p, -663a levels (P<0.050) in microvesicles (0.1μm-1μm in diameter) released by CASMC. Two of these miRNAs, miR-24-3p and miR-130a-3p, were reduced in circulating microvesicles from FH patients compared with normocholesterolemic controls in a pilot study (Study population 1) and in different validation studies (Study populations 1 and 2) (P<0.050). Supporting these results, plasma levels of miR-24-3p and miR-130a-3p were also downregulated in FH patients compared to controls (P<0.050). In addition, plasma levels of miR-130a-3p were inversely associated with coronary atherosclerosis in a cohort of suspected CAD patients (Study population 3) (P<0.050).
Exposure to atherogenic lipoproteins modifies the miRNA profile of CASMC-derived microvesicles and these alterations are reflected in patients with FH. Circulating miR-130a-3p emerges as a potential biomarker for coronary atherosclerosis.
分析致动脉粥样硬化脂蛋白对源自人冠状动脉平滑肌细胞（CASMC）的微泡miRNA特征的影响，并将这些结果应用于家族性高胆固醇血症（FH）和冠状动脉疾病（CAD）患者。
将CASMC暴露于致动脉粥样硬化脂蛋白后收集条件培养基。从两个独立的已确诊FH患者群体和匹配的正常胆固醇血症对照（研究群体1，N = 50；研究群体2，N = 24）以及疑似CAD患者群体（研究群体3，N = 50）中采集血浆样本。使用标准技术分离并鉴定细胞外囊泡。使用RT-qPCR分析一组与血管平滑肌细胞（VSMC）（病理）生理学相关的30种miRNA。
致动脉粥样硬化脂蛋白显著降低了CASMC释放的微泡（直径0.1μm - 1μm）中miR-15b-5p、-24-3p、-29b-3p、-130a-3p、-143-3p、-146a-3p、-222-3p、-663a的水平（P<0.050）。在一项初步研究（研究群体1）以及不同的验证研究（研究群体1和2）中，与正常胆固醇血症对照相比，FH患者循环微泡中的这两种miRNA，即miR-24-3p和miR-130a-3p减少（P<0.050）。支持这些结果的是，与对照相比，FH患者血浆中miR-24-3p和miR-130a-3p的水平也下调（P<0.050）。此外，在一组疑似CAD患者（研究群体3）中，miR-130a-3p的血浆水平与冠状动脉粥样硬化呈负相关（P<0.050）。
暴露于致动脉粥样硬化脂蛋白会改变CASMC衍生微泡的miRNA谱，这些改变在FH患者中得到体现。循环miR-130a-3p成为冠状动脉粥样硬化的潜在生物标志物。
