Table_2_Comprehensive Analysis of circRNA-miRNA-mRNA Regulatory Network and Novel Potential Biomarkers in Acute Myocardial Infarction.XLSX

BackgroundCircular RNA (circRNA) plays an important role in the regulation of gene expression and the occurrence of human diseases. However, studies on the role of circRNA in acute myocardial infarction (AMI) are limited. This study was performed to explore novel circRNA-related regulatory networks in AMI, aiming to better understand the molecular mechanism of circRNAs involvement in AMI and provide basis for further scientific research and clinical decision-making.
MethodsThe AMI-related microarray datasets GSE160717 (circRNA), GSE31568 (miRNA), GSE61741 (miRNA), and GSE24519 (mRNA) were obtained from the Gene Expression Omnibus (GEO) database. After differential expression analysis, the regulatory relationships between these DERNAs were identified by online databases circBank, circInteractome, miRDB, miRWalk, Targetscan, and then two circRNA-miRNA-mRNA regulatory networks were constructed. Differentially expressed genes (DEGs) in this network were selected followed by enrichment analysis and protein–protein interaction (PPI) analysis. Hub genes were identified using Cytohubba plug-in of Cytoscape software. Hub genes and hub gene-related miRNAs were used for receiver operating characteristic curve (ROC) analysis to identify potential biomarkers. The relative expression levels of these biomarkers were further assessed by GSE31568 (miRNA) and GSE66360 (mRNA). Finally, on the basis of the above analysis, myocardial hypoxia model was constructed to verify the expression of Hub genes and related circRNAs.
ResultsA total of 83 DEcircRNAs, 109 CoDEmiRNAs and 1204 DEGs were significantly differentially expressed in these datasets. The up-regulated circRNAs and down-regulated circRNAs were used to construct a circRNA-miRNA-mRNA regulatory network respectively. These circRNA-related DEGs were mainly enriched in the terms of “FOXO signaling pathway,” “T cell receptor signaling pathway,” “MAPK signaling pathway,” “Insulin resistance,” “cAMP signaling pathway,” and “mTOR signaling pathway.” The top 10 hub genes ATP2B2, KCNA1, GRIN2A, SCN2B, GPM6A, CACNA1E, HDAC2, SRSF1, ANK2, and HNRNPA2B1 were identified from the PPI network. Hub genes GPM6A, SRSF1, ANK2 and hub gene-related circRNAs hsa_circ_0023461, hsa_circ_0004561, hsa_circ_0001147, hsa_circ_0004771, hsa_circ_0061276, and hsa_circ_0045519 were identified as potential biomarkers in AMI.
ConclusionIn this study, the potential circRNAs associated with AMI were identified and two circRNA-miRNA-mRNA regulatory networks were constructed. This study explored the mechanism of circRNA involvement in AMI and provided new clues for the selection of new diagnostic markers and therapeutic targets for AMI.

背景：环状RNA（circular RNA, circRNA）在基因表达调控及人类疾病发生发展中发挥重要作用。然而，目前关于环状RNA在急性心肌梗死（acute myocardial infarction, AMI）中作用的研究仍较为有限。本研究旨在探索急性心肌梗死中与环状RNA相关的新型调控网络，以更好地阐明环状RNA参与急性心肌梗死发生的分子机制，为后续相关科学研究及临床决策提供理论依据。
方法：本研究从基因表达综合数据库（Gene Expression Omnibus, GEO）中获取与急性心肌梗死相关的微阵列数据集：GSE160717（circRNA）、GSE31568（微小RNA, miRNA）、GSE61741（miRNA）及GSE24519（信使RNA, mRNA）。完成差异表达分析后，通过circBank、circInteractome、miRDB、miRWalk、Targetscan等在线数据库鉴定上述差异表达RNA（DERNAs）之间的调控关系，并构建两条circRNA-miRNA-mRNA调控网络。选取该网络中的差异表达基因（differentially expressed genes, DEGs）进行富集分析及蛋白质相互作用（protein–protein interaction, PPI）分析。利用Cytoscape软件的Cytohubba插件筛选核心基因（Hub genes）。将核心基因及其关联的miRNA进行受试者工作特征曲线（receiver operating characteristic curve, ROC）分析，以鉴定潜在生物标志物。通过数据集GSE31568（miRNA）与GSE66360（mRNA）进一步评估上述生物标志物的相对表达水平。最后，基于上述分析结果构建心肌缺氧模型，验证核心基因及相关环状RNA的表达情况。
结果：本研究共筛选得到83个差异表达环状RNA（DEcircRNAs）、109个共差异表达miRNA（CoDEmiRNAs）及1204个DEGs。分别以上调表达环状RNA与下调表达环状RNA构建circRNA-miRNA-mRNA调控网络。与环状RNA相关的DEGs主要富集于“FOXO信号通路”“T细胞受体信号通路”“MAPK信号通路”“胰岛素抵抗”“cAMP信号通路”及“mTOR信号通路”等条目。从PPI网络中筛选得到排名前十的核心基因：ATP2B2、KCNA1、GRIN2A、SCN2B、GPM6A、CACNA1E、HDAC2、SRSF1、ANK2及HNRNPA2B1。其中核心基因GPM6A、SRSF1、ANK2以及与其关联的环状RNA hsa_circ_0023461、hsa_circ_0004561、hsa_circ_0001147、hsa_circ_0004771、hsa_circ_0061276、hsa_circ_0045519被鉴定为急性心肌梗死潜在生物标志物。
结论：本研究鉴定出与急性心肌梗死相关的潜在环状RNA，并构建了两条circRNA-miRNA-mRNA调控网络。本研究阐明了环状RNA参与急性心肌梗死发生的潜在机制，为急性心肌梗死新型诊断标志物及治疗靶点的筛选提供了新的研究思路。