DataSheet_1_Serum miRNA Profile in Diabetic Patients With Ischemic Heart Disease as a Promising Non-Invasive Biomarker.docx

The increasing morbidity and mortality of type 2 diabetic mellitus (T2DM) patients with ischemic heart disease (IHD) highlight an urgent need to identify early biomarkers, which would help to predict individual risk of development of IHD. Here, we postulate that circulating serum-derived micro RNAs (miRNAs) may serve as potential biomarkers for early IHD diagnosis and support the identification of diabetic individuals with a predisposition to undergo IHD. We obtained serum samples from T2DM patients either with IHD or IHD-free and analysed the expression levels of 798 miRNAs using the NanoString nCounter technology platform. The prediction of the putative miRNAs targets was performed using the Ingenuity Pathway Analysis (IPA) software. Gene Ontology (GO) analysis was used to identify the biological function and signalling pathways associated with miRNA target genes. Hub genes of protein-protein interaction (PPI) network were identified by STRING database and Cytotoscape tool. Receiver operating characteristic (ROC) analysis was used to assess the diagnostic value of identified miRNAs. Real-time quantitative polymerase chain reaction (qRT-PCR) was used for nCounter platform data validation. Our data showed that six miRNAs (miR-615-3p, miR-3147, miR-1224-5p, miR-5196-3p, miR-6732-3p, and miR-548b-3p) were significantly upregulated in T2DM IHD patients compared to T2DM patients without IHD. Further analysis indicated that 489 putative target genes mainly affected the endothelin-1 signalling pathway, glucocorticoid biosynthesis, and apelin cardiomyocyte signalling pathway. All tested miRNAs showed high diagnostic value (AUC = 0.779 - 0.877). Taken together, our research suggests that circulating miRNAs might have a crucial role in the development of IHD in diabetic patients and may be used as a potential biomarker for early diagnosis.

2型糖尿病（T2DM）合并缺血性心脏病（IHD）患者患病率和死亡率上升，凸显了迫切需要识别早期生物标志物的必要性，以预测个体发生IHD的风险。本研究中，我们提出循环血清来源的微小RNA（miRNA）可能作为早期IHD诊断的潜在生物标志物，并有助于识别具有发生IHD倾向的糖尿病患者。我们收集了T2DM患者（伴有或无IHD）的血清样本，并使用NanoString nCounter技术平台分析了798种miRNA的表达水平。预测假定的miRNA靶标使用了Ingenuity Pathway Analysis（IPA）软件。通过基因本体（GO）分析，确定了与miRNA靶基因相关的生物学功能和信号通路。蛋白质-蛋白质相互作用（PPI）网络的核心基因通过STRING数据库和Cytoscape工具识别。受试者工作特征（ROC）分析被用于评估所识别miRNA的诊断价值。实时定量聚合酶链反应（qRT-PCR）用于验证nCounter平台的数据。我们的数据显示，与无IHD的T2DM患者相比，六种miRNA（miR-615-3p、miR-3147、miR-1224-5p、miR-5196-3p、miR-6732-3p和miR-548b-3p）在T2DM IHD患者中显著上调。进一步分析表明，489个假定的靶基因主要影响内皮素-1信号通路、糖皮质激素生物合成和心肌细胞apelin信号通路。所有测试的miRNA均显示出高诊断价值（AUC = 0.779 - 0.877）。综上所述，我们的研究结果表明，循环miRNA可能在糖尿病患者IHD的发生中发挥关键作用，并可能作为早期诊断的潜在生物标志物。