Screening Of Moyamoya Disease-related MicroRNAs By Transcriptome High-throughput Sequencing As Possible Biomarkers And Joint Analysis

Objective:Moyamoya disease(MMD)is a chronic idiopathic disorder characterized by steno-occlusive lesions around the terminal portions of the internal carotid arteries accompanied by abnormal collateral vessels.The etiology and pathogenesis of this disease are not yet to be clearly illustrated.In order to construct a possible regulatory network for differentially expressed mi RNAs,we screened,validated and bioinformatically analyzed the differential expression profiles of mi RNAs,and further combined the transcriptome sequencing data for association analysis,which may lay a foundation for understanding the molecular biological mechanism of mi RNA in the occurrence and development of MMD.Therefore,it provides possible research directions for early screening of MMD,preventing new cases to the greatest extent and slowing down the disease process through early treatment.Methods: 1.s RNA and transcriptome deep sequencing were performed on whole blood samples of 5 patients with MMD and 5 normal controls by lllumina-Seq technique,and differential expression profiles of mi RNA were analyzed and screened.2.The expression of 7 differentially expressed mi RNAs screened in high-throughput sequencing results were verified by real-time quantitative PCR(RT-q PCR)in the same source sample RT-PCR,while clinical samples were expanded to be 7 cases and 8 controls to further detect the expression changes of 3 interested mi RNAs molecules.3.Target gene prediction of differentially expressed mi RNAs,GO enrichment and KEGG function enrichment were performed using relevant bioinformatics analysis software;4.Through the combined transcriptome sequencing data,the network regulation mechanism of differentially expressed mi RNAs,and the differentially expressed mi RNAs were correlated with m RNA and lnc RNA were explored and analyzed as comprehensively as possible.Results: 1.102 differentially expressed mi RNAs were obtained,41 of which were up-regulated and 61 down-regulated.2.7 mi RNAs with changes in high-throughput sequencing results were selected for verification,the expression of hsa-let-7d-3p were up-regulated and the expression of mi R-210-3p,mi R-126-3p,hsa-let-7f-5p,hsa-let-7b-5p,hsa-mi R-330-5p and mi R-106b-5p were down-regulated.The clinical samples were expanded to be 7 cases and 8 controls.The expression of hsa-let-7d-3p and mi R-210-3p was all up-regulated.3.Prediction of target genes for differentially expressed micro RNAs and functional enrichment analysis showed that a large number of target genes were significantly enriched in cell metabolic processes and mitogen activated protein kinase(MAPK)signaling pathways.4.We have successfully constructed a ce RNA expression regulation network of lnc RNA-micro RNA-RNA,and some of the core micro RNAs deserve our further study.Conclusion: In this study,differentially expressed mi RNAs associated with MMD were obtained and verified by RT-q PCR and bioinformatics analysis.It was confirmed that hsa-let-7d-3p was significantly up-regulated in MMD,which may be involved in the regulation of cell proliferation,migration,apoptosis and angiogenesis,and thus play a role in the pathogenesis and progression of MMD.Through association analysis of transcriptome data,we constructed an expression regulation network with mi RNAs as the core,which laid a foundation for exploring and clarifying the mechanism of the occurrence and development of moyamoya disease at the transcriptome level.