Asulin Nofar, Volinsky Natalia, Grosman-Rimon Liza, Kachel Erez, Sternik Leonid, Raanani Ehud, Altshuler Roman, Magen Iddo, Ben-Zvi Inbar, Margalit Nufar, Carasso Shemy, Meir Karen, Haviv Izhak, Amir Offer
Cardiovascular Department and Research Center, Poriya Medical Center, Tiberias, Israel.
The Azrieli Faculty of Medicine, Bar-Ilan University, Safed, Israel.
J Card Surg. 2020 Jul;35(7):1508-1513. doi: 10.1111/jocs.14636. Epub 2020 Jun 2.
The aortic valve (AV) is the most commonly affected valve in valvular heart diseases (VHDs). The objective of the study is to identify microRNA (miRNA) molecules expressed in VHDs and the differential expression patterns of miRNA in AVs with either calcification or rheumatism etiologies.
Human AVs were collected during valve replacement surgery. RNA was extracted and miRNA containing libraries were prepared and sequenced using the next generation sequencing (NGS) approach. miRNAs identified as differentially expressed between the two etiologies were validated by quantitative real-time polymerase chain reaction (qPCR). The receiver operating characteristic (ROC) curve analysis was performed to examine the ability of relevant miRNA to differentiate between calcification and rheumatism etiologies.
Rheumatic and calcified AV samples were prepared for the NGS and were successfully sequenced. The expression was validated by the qPCR approach in 46 AVs, 13 rheumatic, and 33 calcified AVs, confirming that miR-145-5p, miR-199a-5p, and miR-5701 were significantly higher in rheumatic AVs as compared with calcified AVs. ROC curve analysis revealed that miR-145-5p had a sensitivity of 76.92% and a specificity of 94.12%, area under the curve (AUC) = 0.88 (P = .0001), and miR-5701 had a sensitivity of 84.62% and a specificity of 76.47%, AUC = 0.78 (P = .0001), whereas miR-199a-5p had a sensitivity of 84.62%, and a specificity of 57.58%, AUC = 0.73 (P = .0083).
We documented differential miRNA expression between AV disease etiologies. The miRNAs identified in this study advance our understanding of the mechanisms underlining AV disease.
主动脉瓣（AV）是心脏瓣膜疾病（VHDs）中最常受累的瓣膜。本研究的目的是鉴定在VHDs中表达的微小RNA（miRNA）分子，以及在钙化或风湿病因的主动脉瓣中miRNA的差异表达模式。
在瓣膜置换手术期间收集人主动脉瓣。提取RNA，制备含miRNA的文库，并使用下一代测序（NGS）方法进行测序。通过定量实时聚合酶链反应（qPCR）验证在两种病因之间鉴定为差异表达的miRNA。进行受试者工作特征（ROC）曲线分析，以检验相关miRNA区分钙化和风湿病因的能力。
制备了风湿性和钙化性主动脉瓣样本用于NGS，并成功测序。通过qPCR方法在46个主动脉瓣、13个风湿性主动脉瓣和33个钙化性主动脉瓣中验证了表达，证实与钙化性主动脉瓣相比，miR-145-5p、miR-199a-5p和miR-5701在风湿性主动脉瓣中显著更高。ROC曲线分析显示，miR-145-5p的敏感性为76.92%，特异性为94.12%，曲线下面积（AUC）=0.88（P=0.0001），miR-5701的敏感性为84.62%，特异性为76.47%，AUC=0.78（P=0.0001），而miR-199a-5p的敏感性为84.62%，特异性为57.58%，AUC=0.73（P=0.0083）。
我们记录了主动脉瓣疾病病因之间的miRNA差异表达。本研究中鉴定的miRNA推进了我们对主动脉瓣疾病潜在机制的理解。
