Identification and verification of hub genes associated with ferroptosis in ischemia and reperfusion injury during renal transplantation

Ferroptosis is involved in ischemia and reperfusion injury (IRI) of transplanted kidney. Understanding the molecular mechanisms of ferroptosis is essential to elucidate the pathogenesis of IRI. 1307 differentially expressed genes (DEGs) were obtained by GSE90861 retrieved from the GEO database. 29 ferroptosis-related DEGs were obtained from the intersection with FerrDb database, which were subjected to enrichment analysis and cytoHubba plugin for selecting the top three (IL6, ATF3 and JUN) as hub genes. Next, ROC analysis of hub genes showed good diagnostic prospects in both GSE90861 and GSE126805. Given the close link between ferroptosis and immunity, immunological analysis of CIBERSORTx revealed that the proportions of 10 cell types out of 22 immune cells in the transplanted kidney significantly changed after reperfusion. To study the relationship between IRI and ferroptosis, 15 male C57BL/6j mice were randomly divided into three groups: control (C), ischemia and reperfusion (IR), and IR + Fer-1 (IF) groups. The IRI mouse model not only developed significant histological damage changes, but also exhibited mitochondrial damage, iron accumulation, increased MDA, and decreased GSH. The ferroptosis inhibitor, Fer-1, ameliorated renal IRI, as demonstrated by rise of GPX4 and decline of TFRC, PTGS2 and ACSL4. In addition, hub genes were further confirmed by significant increase in IRI mouse model the same as the GEO database. In brief, ferroptosis-related hub genes (IL-6, ATF3 and JUN) screened were closely relevant to immune response and might be diagnostic biomarkers and therapeutic targets for IRI during renal transplantation, which could prevent renal allograft dysfunction.