Screening Of Differentially Expressed CircRNAs In IgA Nephropathy And Bioinformatics Analysis Of Hsa?circ?0073237

Objective:IgA nephropathy(IgAN),an immune-mediated primary glomerulonephritis,is the most common type of primary glomerulonephritis worldwide and a leading cause of end-stage renal disease in young adults.However,its pathogenesis remains unclear.More and more evidences show that circRNA can act as competitive endogenousRNAs(ceRNA)interacting with miRNAs,which has attracted more and more attention in the field of renal disease.However,the role of circRNA in IgAN is still unclear.The purpose of this study was to explore the potential circRNA and downstream microRNA/mRNA pathways in IgAN and to provide a new research direction for the genetic and molecular mechanisms of IgAN.Methods:1.Download high-throughput sequencing data GSE154046 from the GEO database.R language was used to run edger and limma package and analyze the difference of the data,identifying the differentially expressed circRNAs and screening the significantly differentially expressed circRNAs.The first three circRNAs with the most significant differential expression "hsa?circ?0073237?hsa?circ?0007720?hsa?circ?0073239" were selected for subsequent analysis.2.The relevant information of three circRNAs was obtained by circ Base(circbase.org).Then the basic structure diagram of differentially expressed circRNAs was made by using CSCD database.The Circinteractome database and CSCD database were used to predict the downstream microRNA of the three circRNAs above.The Perl Programming Language was used to obtain the intersection.Cytoscape software was used to construct the circRNA-miRNA network diagram.3.The analysis of miRNA-mRNA was performed for the most significant circRNA,i.e.,hsa?circ?0073237 corresponding to the minimum adjust P Value.The downstream target genes of miRNAs acting as ceRNA with hsa?circ?0073237 were predicted through three databases,mi RDB,mi RTARbase and Target Scan Human.The intersection of the three databases is taken as the final prediction result.Finally,the miRNA-mRNA network was visualized using Cytoscape software.4.Cluster Prolifer template was run by R language software to perform GO and KEGG enrichment analysis on the prediction results of target genes.5.Use STING Version 11.0 to make protein-protein interaction network then use the Perl programming language to predict the core genes of the network.Results:1.After the difference analysis of GSE154046 data set obtained from the GEO database,significantly differentially expressed circRNAs were obtained.An analysis of15,070 circRNAs between IgAN and healthy controls showed that 11,622 circRNAs were up-regulated and 3,448 circRNAs were down-regulated.Then use R language to run the edger and limma difference analysis package,with conditions "| log FC | > 1,adj.P.v al< 0.05" for the screening of significantly differentially expressed circRNAs.The results showed that there were 26 circRNAs,of which 14 were up-regulated and 12 were down-regulated.2.Prediction results of downstream miRNAs were obtained.The first three circRNAs with the most significant differential expression were hsa?circ?0073237,hsa?circ?0007720 and hsa?circ?0073239.Circ Base database was used to obtain the sequence,chromosome position and structure schematic diagram of the three circRNAs.Circinteractome database and CSCD database were performed to predict downstream microRNA of the three circRNAs.There were 48 miRNAs interacting with hsa?circ?0073237,44 miRNAs interacting with hsa?circ?0007720 and 44 miRNAs interacting with hsa?circ?0073239.3.The mRNA prediction results of hsa?circ?0073237/miRNA downstream target genes were obtained by using three databases,mi RDB,mi Tarbase and Target Scan Human.The miRNA-mRNA network was made by Cytoscape software.4.GO and KEGG enrichment analysis was performed on the prediction results of target genes by running the Clusterprolifer template using R language.The results suggested that hsa?circ?0073237-miRNA-mRNA network is involved in the biogenic processes such as transcription co-regulator activity,DNA-binding transcription factor binding,transcription coactivator protein activity,translation regulator activity,and regulation of protein kinase regulator.It is involved in autophagy,HIF-signaling pathway,Wnt signaling pathway,erb B signaling pathway and other pathways.5.The STRING Version 11.0 was used to make the protein-protein interaction network,and it was found that there were interaction relationships between multiple target gene mRNAs and mRNAs in the hsa?circ?0073237-miRNA-mRNA axis.The results were displayed by the PPI protein network,and the target gene with the largest number of protein-protein interaction nodes was MYC after the prediction of the core genes in the network.Based on the above bioinformatics analysis,it can provide a potential new idea for the study of the pathogenesis of IgA nephropathy.Conclusion:CircRNAs play a potentially mediating role in the pathogenesis and development of IgA nephropathy through the ceRNA mechanism.Hsa?circ?0073237-miRNA-mRNA plays a potentially important role in the pathogenesis of IgAN.