To Analyze The Ferroptosis Genes And Potential Drugs Associatted With Immune Infiltration In Hashimoto’s Thyroiditis By Bioinformatics

Background and objective: HT is one of the most common autoimmune diseases,which is characterized by thyroid-specific autoantibodies.Although the exact etiology has not been fully elucidated,Hashimoto’s thyroiditis is associated with genetic factors,environmental factors,and autoimmune interactions.A new type of iron-mediated cell death(ferroptosis)has also been reported to be associated with the initiation of inflammation and increased immune infiltration.Recent studies have found that Ferroptosis and oxidation share several features,such as lipoxygenase action,ROS production,and gene expression.Ferroptosis differs from apoptosis,necrosis,and pyroptosis in morphological and physiological characteristics.During ferroptosis,the nucleus remains intact.Chromatin was not aggregated,and the plasma membrane was not ruptured or blebbing.The contracted mitochondria showed a greater inner membrane density while the outer membrane was ruptured.The most important biochemical features of Ferroptosis are elevated concentrations of lipid hydroperoxides(LOOH)and ferrous ions(Fe2+),since Ferroptosis cells produce excess reactive oxygen species that initiate lipid peroxidation.Glutathione peroxidase4(GPX4),an enzyme that specifically reduces lipid peroxides to the corresponding alcohol,is another central regulator of ferroptosis.In addition,glutathione(GSH),as a cofactor of GPX4,maintains the level of GPX4 by exchanging glutamate and cystine through the reverse transport system Xc.The genes that control Ferroptosis are also distinct from those that control other forms of cell death.SHRNA libraries were used to target genes encoding predicted mitochondrial proteins,including those encoding ribosomal protein L8(RPL8),iron response element binding protein 2(IREB2),and ATP synthase F0.Six protein-coding genes,complex subunit C3(ATP5G3),citrate synthase(CS),tetrapeptides repeat domain 35(TTC35),and acyl-Co A synthetase family member 2(ACSF2),required for ferroptosis death were screened in HT1080 and Ca LU-1 cells.In addition,TFRC,ISCU,FTH1,and FTL are key genes for ferroptosis,controlling iron uptake,metabolism,and storage by affecting FE 2+ levels.These genes are distinct from genes that control apoptosis,such as BH3-interacting domain death agonist(BID),BCL2 antagonist/killer 1(BAK1),Bcl2-associated X(BAX),apoptosis-inducing factor mitochondria-associated 1(AIFM1),or control other modes of cell death,such as involvement in MPT The gene driving necrosis,peptidyl-prolyl isomerase F(PPIF).The aim of this study is to investigate the expression of ferroptosis-related genes in Hashimoto’s thyroiditis,its role in immune infiltration,and potential therapeutic targets.Methods: The dataset GSE138198 was downloaded to identify and analyze differentially expressed genes(DEGs)between Hashimoto’s thyroiditis patients and healthy control samples.GO and KEGG enrichment analysis was used to analyze the functions of the DEGs.The DEGs were intersected with ferroptosis-related genes(FRGs)from The Ferr Db database to obtain differentially expressed FRGs(DEFRGs).A protein-protein interaction network(PPI)was established by Cytoscape software to identify hub genes.Real-time polymerase chain reaction(RT-PCR)was used for the validation of hub genes.Gene set enrichment analysis(GSEA)was also conducted for each hub gene.Then immune infiltration characteristics were analyzed by CIBERSORT.Furthermore,potential therapeutic drug were identified using the Connectivity Map(CMAP)database and validated by RT-PCR.Results: A total of 75 differentially expressed ferroptosis-related genes(24upregulated genes and 51 down-regulated genes)between Hashimoto’s thyroiditis patients and healthy controls were identified.Based on the PPI network,9 hub genes were screened out.The expressions of 7 hub genes were consistent in RT-PCR validation and DEFRGs data.The immunological features of Hashimoto’s thyroiditis samples had an increase in memory B cells and M1 macrophages and a decrease in M0 macrophages,monocytes,activated dendritic cells,activated NK cells,and neutrophils.At the same time,the above performance is related to the differential expression of the hub gene.The hub gene was verified in the collected clinical samples of Hashimoto’s thyroiditis patients and healthy controls by q PCR.Through the CMAP database,we found that the compound MLN4924 could be a potential therapeutic drug for Hashimoto’s thyroiditis.And we finally validated the effect of MLN4924 on Hub genes in Hashimoto’s thyroiditis cell model stimulated by TNF-αand IFN-γ.Conclusion: This study provides a deeper understanding of the molecular pathogenesis of Hashimoto’s thyroiditis.We found that ferroptosis plays an essential role in Hashimoto’s thyroiditis by affecting immune infiltration.And ferroptosis-related genes may be potential therapeutic targets for Hashimoto’s thyroiditis.It is suggested that the compound MLN4924 can be further explored as a potential medicinal drug for Hashimoto’s thyroiditis.