Bioinformatics-based screening of potential key targets of ferroptosis involved in multiple sclerosis mechanisms and preliminary validation
Objective To screen the potential key genes in multiple sclerosis(MS)from the perspective of fer-roptosis by bioinformatics technology,and to explore the mechanism of ferroptosis in MS by preliminary verifica-tion in animal experiment.Methods We downloaded MS datasets GSE21942 and GSE43591 from the GEO data-base and obtained ferroptosis-related genes from the FerrDb database and related literature.The dataset GSE21942 was analyzed using R software for differential analysis and WGCNA analysis.The intersection of the results from both analyses and the ferroptosis-related genes yielded the MS ferroptosis-related differentially ex-pressed genes(FRDEGs).Subsequently,GO and KEGG enrichment analyses were performed on the FRDEGs.Two machine learning algorithms,LASSO and SVM-RFE,were employed to further screen the key genes,which were validated using the dataset GSE43591,and the diagnostic value of the key genes was assessed by plotting ROC curves.The ssGESA algorithm was used to explore the correlation between key genes and immune cells.Fi-nally,the expression of key genes was further validated using qPCR and Western blot methods.Results A total of 20 FRDEGs were screened.The results of GO and KEGG enrichment analysis indicated that they were mainly in-volved in response to oxidative stress,negative regulation of protein phosphorylation,participation in the NOD-like receptor signaling pathway,MAPK signaling pathway,Th17 cell differentiation,etc.Machine learning and validation with external datasets screened two key genes(CIRBP and TNFAIP3)related to ferroptosis.The ROC results indicated that both CIRBP and TNFAIP3 had high diagnostic values.The results of immune infiltration a-nalysis showed that CIRBP and TNFAIP3 were significantly associated with activated CD4 T cells and activated B cells.qPCR and Western blot assays showed that the mRNA and protein expression levels of CIRBP and TN-FAIP3 were significantly increased in mice of the EAE group compared with those of the normal group(P＜0.05).Conclusion CIRBP and TNFAIP3 are potential key genes associated with ferroptosis in MS,and the in-volvement of ferroptosis in the pathogenesis of MS may be related to the high expression of CIRBP and TNFAIP3.
万方数据
维普
