Identification of ferroptosis-related genes in type 2 diabetes mellitus based on machine learning

BackgroundType 2 diabetes mellitus (T2DM), which has a high incidence and several harmful consequences, poses a severe danger to human health. Research on the function of ferroptosis in T2DM is increasing. This study uses bioinformatics techniques identify new diagnostic T2DM biomarkers associated with ferroptosis.MethodsTo identify ferroptosis-related genes (FRGs) that are differentially expressed between T2DM patients and healthy individuals, we first obtained T2DM sequencing data and FRGs from the Gene Expression Omnibus (GEO) database and FerrDb database. Then, drug-gene interaction networks and competitive endogenous RNA (ceRNA) networks linked to the marker genes were built after marker genes were filtered by two machine learning algorithms (LASSO and SVM-RFE algorithms). Finally, to confirm the expression of marker genes, the GSE76895 dataset was utilized. The protein and RNA expression of some marker genes in T2DM and nondiabetic tissues was also examined by Western blotting, immunohistochemistry (IHC), immunofluorescence (IF) and quantitative real-time PCR (qRT-PCR).ResultsWe obtained 58 differentially expressed genes (DEGs) associated with ferroptosis. GO and KEGG enrichment analyses showed that these DEGs were significantly enriched in hypoxia and ferroptosis. Subsequently, eight marker genes (SCD, CD44, HIF1A, BCAT2, MTF1, HILPDA, NR1D2, and MYCN) were screened by LASSO and SVM-RFE machine learning algorithms, and a model was constructed based on these eight genes. This model also has high diagnostic power. In addition, based on these eight genes, we obtained 48 drugs and constructed a complex ceRNA network map. Finally, Western blotting, IHC, IF, and qRT-PCR results of clinical samples further confirmed the results of public databases.ConclusionsThe diagnosis and aetiology of T2DM can be greatly aided by eight FRGs, providing novel therapeutic avenues. In this study, we identified eight hub genes (SCD, CD44, HIF1A, BCAT2, MTF1, HILPDA, NR1D2, and MYCN) that are closely associated with ferroptosis in T2DM. The three ferroptosis genes (HIF1A, HILPDA, and SCD) are strongly related to T2DM, hypoxia and lipid metabolism, providing new research directions for the development and treatment of T2DM and its complications. Based on these eight genes, we constructed a model with a high ability to diagnose T2DM. We also predicted the drugs corresponding to these eight genes as well as constructed a ceRNA network map. In addition, we verified the protein expression of CD44 and MYCN in T2DM and nondiabetic tissues by Western blotting, Immunohistochemistry staining, Immunofluorescence Staining and quantitative real-time PCR and the results were statistically significant. The above findings suggest that further studies of ferroptosis may offer new therapeutic goals and biomarkers for patients with T2DM.image