影响扩张型心肌病干细胞移植疗效的差异miRNA表达分析  

Analysis of differential miRNA expression related to the efficacy of stem cell transplantation in dilated cardiomyopathy

作　　者：范家铭 杨秭莹 冯振辉 陈一欢[1] 王雨桐 沈振亚[1] Fan Jiaming;Yang Ziying;Feng Zhenhui;Chen Yihuan;Wang Yutong;Shen Zhenya(Department of Cardiovascular Surgery of the First Affiliated Hospital of Soochow University,Suzhou 215006,China)

机构地区：[1]苏州大学附属第一医院心脏大血管外科,215006

出　　处：《中华细胞与干细胞杂志（电子版）》2023年第5期288-298,共11页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：国家自然科学基金资助项目(92168203);博习培育计划项目(BXQN2023006,202131);江苏省科研项目(M2022010,21KJB310003);苏州市医学研究项目(SKY2022131,KJXW2021001)。

摘　　要：目的基于生物信息学分析探索影响扩张型心肌病(DCM)干细胞移植疗效的差异表达微小RNA(miRNAs)及其靶基因。方法本研究筛选术前基线特征相似的DCM患者,以干细胞移植术后左心室射血分数改善>5%为标准,分为治疗良好组和无效组。通过对两组患者的骨髓间充质干细胞样本进行miRNAs高通量测序,运用edgeR筛选出两组间的差异表达miRNAs,并使用MiRanda、TargetScan和RNAhybrid数据库进行靶基因预测,对预测得到的靶基因进行GO、KEGG及Reactome富集分析。使用STRING数据库和Cytoscape软件,并参考紧密度、介数中心性、度三种拓扑分析方法和韦恩图筛选关键基因。两组间比较采用t检验。结果本研究共筛选出44个差异表达miRNAs,其中23个上调,21个下调。上调的差异表达miRNAs靶基因有908个,下调的差异表达miRNAs靶基因835个。GO分析结果显示靶基因在生物过程中主要富集在细胞代谢过程的调节,分子功能主要富集在离子结合,细胞组分则主要与细胞膜相关。KEGG通路和Reactome分析发现靶基因主要与调节干细胞多能性的信号通路、RIG-I样受体信号通路和RNA聚合酶II转录等相关。此外,韦恩图共筛选出4个关键基因,分别为MED1、SMAD2、TRAF6、GSK3B,其中GSK3B和TRAF6分别为上调的hsa-miR-1304-3p和下调的hsa-miR-146a-3p靶基因,MED1和SMAD2为下调的hsa-miR-1299靶基因。结论本研究提供了hsa-miR-1304-3p、hsa-miR-1299、hsa-miR-146a-3p 3个可能用于提示DCM患者干细胞移植预后的差异表达miRNAs,其调控的关键基因MED1、SMAD2、TRAF6、GSK3B及所在通路可能是提高干细胞移植治疗DCM的潜在作用靶点。Objective To identify differentially expressed miRNAs(DEmiRNAs)and their targets that potentially influence the therapeutic efficacy of bone marrow mesenchymal stem cells(BMSCs)transplantation for dilated cardiomyopathy(DCM).Methods The DCM patients with similar baseline characteristics before stem cell transplantation were selected and categorized into two groups based on the improvement of left ventricular ejection fraction after stem cell transplantation,namely response group(improvement>5%)and nonresponse group(improvement≤5%).High-throughput sequencing of miRNAs in BMSCs samples from two groups was performed,and DEmiRNAs were analyzed by the edgeR tool.Furthermore,the target genes of DEmiRNAs were predicted by MiRanda,TargetScan,and RNAhybrid tools,and subjected to GO,KEGG,and Reactome enrichment analyses.Finally,utilizing the STRING database and Cytoscape software,three topological analysis methods based on closeness,betweenness,and degree,and the Venn diagram were employed to identify the hub genes.Results This study identified 44 DEmiRNAs,including 23 upregulated and 21 downregulated.908 and 835 genes were predicted as target genes for upregulated and downregulated miRNAs respectively.GO analyses indicated that the target genes were mainly enriched in the regulation of cell metabolism in biological processes(BP),ion binding in molecular functions(MF),and cell membrane in cell components(CC).In addition,KEGG and Reactome pathway analyses discovered signal pathways regulating stem cell pluripotency,RIG-I-like receptor,and RNA polymerase II transcription as the most enriched pathways.Moreover,MED1,SMAD2,TRAF6,and GSK3B were identified as hub genes,and GSK3B and TRAF6 were the target genes of upregulated hsa-miR-1304-3p and downregulated hsa-miR-146a-3p respectively.MED1 and SMAD2 were the target genes of downregulated hsa-miR-1299.Conclusion The study predicted three DEmiRNAs that potentially influence the prognosis of DCM patients after stem cell transplantation.The hub genes MED1,SMAD2,TRAF6,and GSK3B an

关 键 词：扩张型心肌病 间充质干细胞 MICRORNAS 基因表达分析 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R542.2[医药卫生—基础医学]