ارزيابي SLCO5A1 ، miR-218-5p و PILRB به عنوان بيوماركرهاي جديد براي تشخيص بيماري مالتيپل اسكلروزيس با استفاده از سيستم بيولوژي و Real-Time PCR

eva‎luation of SLCO5A1, miR-218-5p and PILRB as new biomarkers for the diagnosis of multiple sclerosis using systems biology and Real-time PCR

زيست شناسي سلولي مولكولي و ميكروبيولوژي

Introduction: Multiple sclerosis (MS) is an inflammatory demyelinating disease of the human central nervous system. The clinical course of the disease is variable, usually starting with reversible episodes of neurological disability in the third or fourth decade of life and becoming a persistent and irreversible neurodegenerative disease by the sixth or seventh decade. Differences in gene expression patterns have been recorded not only in multiple sclerosis patients versus healthy individuals, but also in disease recurrence. In this study, using systems biology tools, genes with different expression patterns were collected between people with MS and healthy people, and finally bioinformatics analysis led to the construction of a new mRNA-miRNA-lncRNA regulatory network involved in the occurrence of MS. Materials and Methods: First, in the bioinformatics phase, massive RNA-seq data related to MS and normal were downloaded, and then the data were analyzed using bioinformatics tools in Linux platform. After examining the differential expression of genes with DESeq2, the co-expression network was reconstructed by WGCNA tool; Finally, the key genes in this disease were identified. To investigate the biological functions of the identified key genes, biological pathway analysis was performed by Enrichr and GSEA. Several online databases MIRMAP, mirDB, targetscan, mirDIP, DIANA Tools and mirwalk were used to predict the upstream miRNAs of the desired gene. Potential upstream lncRNAs that interacted with key miRNAs were predicted using the Starbase online database. In the laboratory phase, the expression of all three biomarkers was measured by Real-Time PCR between two MS and normal groups. Results: Examining the differential expression of genes showed that 4156 genes had significant expression changes in disease state compared to normal. In the co-expression network analysis, the mediumorchid module was selected as the key module in the occurrence of the disease. Finally, from the sharing between the analysis of the differential expression of genes and the co-expression network and the investigation of the functional pathways, the SLCO5A1 gene was selected as the key gene and by checking the online databases that predict the upstream miRNA of the desired gene, has-miR-218-5p was selected as the key miRNA and finally, STAG3L5P-PVRIG2P-PILRB was selected as the key lncRNA with starbase database. With the laboratory investigations of SLCO5A1 in MS disease compared to the healthy state, the significant increase in expression (log2FC=2.27, p value<0.0001), STAG3L5P-PVRIG2P-PILRB, also in the disease state, the significant increase in expression compared to the healthy state (log2FC=7.48, p value< 0.0001) and has-miR-218-5p showed a significant decrease in expression in the disease state compared to the healthy state (log2FC=0.041, p value<0.0001). Conclusion: A novel ceRNA network was successfully constructed, each component significantly associated with MS disease prognosis.

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