Differential Analysis Of CircRNA Gene Expression In Brain Tissue Of Children With Tuberous Sclerosis Epilepsy And Normal Children

Objectives To investigate the differential expression profiles and biological functions of circRNAs in children’s TSC epilepsy cortical nodules and children’s normal cerebral cortex,and to explore new clinical markers and potential therapeutic targets of circRNAs.Methods 5 brain tissue samples from diagnosed children with TSC epilepsy and 5 brain tissue samples from normal children were used for circRNAs high-throughput sequencing,and the differences in the expression of circRNAs in the brain tissues of the two groups were detected and analyzed.Taking the fold change(FC)≥ 2 and P< 0.05 as the screening conditions,GO enrichment and KEGG pathway analysis were performed on the differentially expressed circRNAs in the two groups of samples.mi Randa was used to predict the potential binding mi RNAs sites of significantly differentially expressed circRNAs,to construct a circRNAmi RNA-m RNA gene expression regulatory network,and to infer potential biological functions based on bioinformatics analysis and prediction results.q-PCR was used to verify the relative expression levels of cortical nodules in children with TSC epilepsy and in children with normal cerebral cortex.Results The 5 children with TSC epilepsy were all diagnosed with tuberous sclerosis epilepsy,4 boys and 1 girl,all had epileptic seizures without obvious incentives;the average disease duration was 3.90 years(1.67-6.00 years),and the surgical age was 4.33 years.age(1.92～6.25 years old).Using FC≥2 and P<0.05 as the differential expression criteria,a total of 517 circRNAs differentially expressed in children with TSC epilepsy and normal children were screened,of which 312 circRNAs were up-regulated and 205 were down-regulated.From the results of bioinformatics GO functional analysis,it was found that most of these differentially expressed circRNAs are located in dendritic branches,nuclear inclusion bodies,and presynaptic active regions,and have the functions of regulating 1-phosphatidylinositol 3--kinase activity and participating in hydrolysis.Positive regulation of enzyme activity,hematopoiesis,immune system development,embryonic organ development and other biological processes.Disease enrichment found that these differentially expressed circRNAs were mostly associated with major depression and multiple sclerosis,including epilepsy.The results of KEGG pathway enrichment analysis indicated that the linear genes corresponding to circRNAs and mi RNA-m RNA were involved in the PI3K-Akt and m TOR signaling pathways closely related to TSC epilepsy.The circRNAs-mi RNA co-expression and interaction analysis showed that these differentially expressed circRNAs had multiple mi RNAs binding sites.Among them,hsa-mi R-6511a-3p,hsa-mi R-6511b-3p,hsa-mi R-671-5p,17＿30568-3p(dsimi R-983a-3p)can regulate GNB4,while hsa-mi R-6772-3p and 16＿28944-3p(mmu-mi R-6943-3p)can regulate ITGB4,while GNB4 and ITGB4 are located upstream of PI3K-Akt-m TOR signaling pathway and have direct regulatory activation.Through the analysis of mi RNAm RNA regulation,it was found that hsa-mi R-671-5p was significantly affected by EIF5 B,hsami R-671-5p and hsa-mi R-6772-3p were significantly affected by MCM5,hsa-mi R-671-5p and hsa-mi R-6511a-3p may have a negative regulatory effect on NMT1.q-PCR verification found that in addition to hsa＿circ＿6431,the remaining four hsa＿circ＿0010716(P=0.0235),hsa＿circ＿7697(P=0.0169),hsa＿circ＿0010716(P=0.0235),hsa＿circ＿7697(P=0.0169)among the five circRNAs related to PI3K-Akt and m TOR signaling pathways in the cerebral cortex of children with TSC epilepsy and normal children,The relative m RNA expression levels of hsa＿circ＿7707(P=0.0217)and hsa＿circ＿9929(P=0.0217)were significantly lower than those of normal children,and the difference was statistically significant.Conclusions In this study,we determined the differential expression profile of circRNAs in the brain tissues of children with TSC epilepsy.Bioinformatics analysis indicated that these differentially expressed circRNAs may play an important regulatory role in the development of TSC epilepsy in children through the circr Na-mir Na-MTOR regulatory network.Figure 8;Table 7;Reference 141...