Ortega Ana, Tarazón Estefanía, Gil-Cayuela Carolina, García-Manzanares María, Martínez-Dolz Luis, Lago Francisca, González-Juanatey José Ramón, Cinca Juan, Jorge Esther, Portolés Manuel, Roselló-Lletí Esther, Rivera Miguel
Cardiocirculatory Unit, Health Research Institute La Fe, Valencia, Spain.
Center for Biomedical Research Network in Cardiovascular Diseases (CIBERCV), Madrid, Spain.
PLoS One. 2017 Sep 21;12(9):e0185062. doi: 10.1371/journal.pone.0185062. eCollection 2017.
Alterations in myocardial structure and reduced cardiomyocyte adhesions have been previously described in dilated cardiomyopathy (DCM). We studied the transcriptome of cell adhesion molecules in these patients and their relationships with left ventricular (LV) function decay. We also visualized the intercalated disc (ID) structure and organization. The transcriptomic profile of 23 explanted LV samples was analyzed using RNA-sequencing (13 DCM, 10 control [CNT]), focusing on cell adhesion genes. Electron microscopy analysis to visualize ID structural differences and immunohistochemistry experiments of ID proteins was also performed. RT-qPCR and western blot experiments were carried out on ID components. We found 29 differentially expressed genes, most of all, constituents of the ID structure. We found that the expression of GJA3, DSP and CTNNA3 was directly associated with LV ejection fraction (r = 0.741, P = 0.004; r = 0.674, P = 0.011 and r = 0.565, P = 0.044, respectively), LV systolic (P = 0.003, P = 0.003, P = 0.028, respectively) and diastolic dimensions (P = 0.006, P = 0.001, P = 0.025, respectively). Electron microscopy micrographs showed a reduced ID convolution index and immunogold labeling of connexin 46 (GJA gene), desmoplakin (DSP gene) and catenin α-3 (CTNNA3 gene) proteins in DCM patients. Moreover, we observed that protein and mRNA levels analyzed by RT-qPCR of these ID components were diminished in DCM group. In conclusion, we report significant gene and protein expression changes and found that the ID components GJA3, DSP and CTNNA3 were highly related to LV function. Microscopic observations indicated that ID is structurally compromised in these patients. These findings give new data for understanding the ventricular depression that characterizes DCM, opening new therapeutic perspectives for these critically diseased patients.
先前已报道扩张型心肌病(DCM)存在心肌结构改变和心肌细胞黏附减少的情况。我们研究了这些患者中细胞黏附分子的转录组及其与左心室(LV)功能衰退的关系。我们还观察了闰盘(ID)的结构和组织。使用RNA测序分析了23个移植的左心室样本的转录组谱(13个DCM样本,10个对照[CNT]样本),重点关注细胞黏附基因。还进行了电子显微镜分析以观察ID结构差异以及ID蛋白的免疫组织化学实验。对ID成分进行了RT-qPCR和蛋白质印迹实验。我们发现了29个差异表达基因,其中大多数是ID结构的组成成分。我们发现GJA3、DSP和CTNNA3的表达与左心室射血分数直接相关(分别为r = 0.741,P = 0.004;r = 0.674,P = 0.011;r = 0.565,P = 0.044),与左心室收缩期(分别为P = 0.003,P = 0.003,P = 0.028)和舒张期尺寸(分别为P = 0.006,P = 0.001,P = 0.025)也直接相关。电子显微镜照片显示DCM患者的ID卷曲指数降低,连接蛋白46(GJA基因)、桥粒斑蛋白(DSP基因)和连环蛋白α-3(CTNNA3基因)蛋白的免疫金标记减少。此外,我们观察到通过RT-qPCR分析的这些ID成分的蛋白质和mRNA水平在DCM组中降低。总之,我们报告了显著的基因和蛋白质表达变化,并发现ID成分GJA3、DSP和CTNNA3与左心室功能高度相关。显微镜观察表明这些患者的ID在结构上受损。这些发现为理解DCM特征性的心室功能减退提供了新数据,为这些重症患者开辟了新的治疗前景。