Differential MRNA-miRNA-circRNA Expression And Association Analysis Of Moyamoya Disease Based On High Throughput RNA Sequencing

Objective:Moyamoya disease(MMD),also known as abnormal cerebral reticular vascular disease,is a chronic cerebral vascular occlusive disease of unknown etiology.In order to understand the characteristics of the circular RNA(circRNA)expression profile of moyamoya disease,we applied lllumina-Seq technology to deep transcriptome sequencing of whole blood from moyamoya disease patients.Screening and verification of differentially expressed circRNAs between patients with moyamoya disease and healthy controls,bioinformatics analysis of high-throughput results,and further correlation analysis of the results of the combined transcriptome sequencing data to construct a possible ceRNA regulatory network for differentially expressed circRNAs.It lays the foundation for understanding the molecular biological mechanism of circRNA in moyamoya disease.Methods: 1.Use lllumina-Seq technology to perform sRNA and transcriptome deep sequencing on whole blood samples of 5 patients with moyamoya disease patients and 5 healthy controls,analyze and screen circRNA differential expression profiles;2.Select circRNA for further expansion of sample size verification.In 8 patients with moyamoya disease and 7 normal controls,the expression levels of the six circRNAs screened in the expanded samples were detected by real-time quantitative PCR;3.using software to perform bioinformatics analysis of the high-throughput results,Perform GO enrichment and KEGG functional enrichment analysis of differentially expressed circRNA-derived genes;4.Combine transcriptome sequencing data to analyze the differentially expressed circRNA,miRNA,and mRNA,and draw the core ceRNA regulation of circRNA in the transcriptome The internet.Results: 1.This study used lllumina-Seq technology to obtain differential expression profiles of circRNA related to moyamoya disease.A total of 51 genes were found to be differentially expressed,of which 23 circRNAs were up-regulated and 28 circRNAs were down-regulated.2.Through RT-qPCR verification of whole transcriptome sequencing samples and independent enlarged samples,the results showed that hsa?circ?0005946,hsa?circ?0007079,hsa?circ?0001360,hsa?circ?0002490,and hsa?circ?0014606 showed a clear upward regulation trend;circRNA-HIPK3 showed a downward regulation trend.KO enrichment and KEGG enrichment were performed on the genes of differentially expressed circRNAs.The GO enrichment analysis results showed that most of the differentially expressed circRNAs were enriched into the biological processes related to cell metabolic regulation.The KEGG-enriched pathway is the PI3K-Akt signaling pathway,suggesting that it may play a central role in the pathogenesis of moyamoya disease.4.Based on the ceRNA theory,draw a circRNA-miRNA-mRNA ceRNA regulatory network based on the differentially expressed circRNA,miRNA,and differentially expressed mRNAs.By visualizing the relationship between the three RNAs,narrowing the scope of the study,making circRNAs in moyamoya disease The continued exploration of the specific role mechanism provided provided a theoretical basis.Conclusion: In this study,the differential expression profile of circRNA related to moyamoya disease was obtained by lllumina-Seq technology.Through RT-qPCR and biological analysis,it was proved that circRNA-HIPK3 may participate in the development of moyamoya disease by regulating angiogenesis.The PI3K-AKT signaling pathway is the most relevant research pathway discovered for moyamoya disease,and may be the core pathway of the disease.At the same time,we combined the transcriptome data to draw the circRNA-miRNA-mRNA regulatory network map of ceRNA.The ceRNA regulatory network map is useful for our research in discovering and screening circRNA and ceRNA networks that are at the core of network regulation.It can further narrow the scope of the study and provide further research on the function and transcriptomics of circRNA related to moyamoya The study provides a theoretical basis.