Identification of Immune-Related Ferroptosis Biomarkers in Diabetic Kidney Disease and Screening of Associated Inhibitors.

OBJECTIVE

Immune infiltration and ferroptosis play pivotal roles in the progression of diabetic kidney disease (DKD). However, investigations of immune cell-related ferroptosis genes (ICRFGs) in the context of DKD are insufficient. This study aimed to identify ICRFGs relevant to DKD and screen related inhibitors.

METHODS

In this study, two DKD datasets from the GEO database were utilized. We adopted the ESTIMATE algorithm to generate microenvironment scores. The CIBERSORT and WGCNA methods were employed to identify immune-related differentially expressed genes (DEGs). The common ICRFGs were derived through a Venn diagram. We employed random forest, LASSO, K-M survival, receiver operating characteristic (ROC) curve, clinical relevance, and Spearman correlation analyses to select hub ICRFGs further. Immunohistochemical experiments were also performed to validate the expression. Additionally, we utilized the Selleck database to obtain ferroptosis-related compounds and used USCF Chimera 1.14 to minimize energy, combined with molecular dynamics (MD) simulations to explore possible ferroptosis inhibitors.

RESULTS

Immunohistochemical analysis revealed that arachidonate 5-lipoxygenase (ALOX5) was significantly highly expressed in the db/db group. Clinical correlation and K-M survival analyses confirmed ALOX5 as the most crucial ICRFG in DKD. Furthermore, ALOX5 was significantly enriched in the terms ECM-receptor interaction, regulation of chemokine production, and regulation of the inflammatory response. A positive correlation was observed between ALOX5 and M1 macrophages, γδ T cells, and monocytes. Moreover, virtual screening and MD revealed NSC348884, salvianolic acid B, and deltarasin as potential ferroptosis inhibitors in combination with ALOX5.

CONCLUSION

We identified ALOX5 as a reliable and prospective diagnostic marker associated with immunity and ferroptosis in DKD patients.