Identification of ferroptosis-related molecular markers in glomeruli and tubulointerstitium of lupus nephritis.

ABSTRACT

PURPOSE: Ferroptosis, characterized by iron accumulation and lipid peroxidation, is a newly demonstrated form of programed cell death. Present studies reveal that ferroptosis is involved in tumor and neurodegenerative disease. Regarding its roles in the development of LN, it is least interrogated. In this study, we explored whether ferroptosis is activated and how does it change at transcriptomic level in LN. METHODS: 4-Hydroxynonenal (4-HNE) was stained to explore whether ferroptosis is activated. Subsequently, by using bioinformatic methods, public GSE32591 dataset was analyzed. Ferroptosis-related differentially expressed genes (FR-DEGs) were identified in both glomeruli and tubulointerstitium. Immune cell infiltration was evaluated. Correlation between FR-DEGs and infiltrated immune cells was also calculated. Finally, dataset of GSE113342, qPCR, and immunofluorescence staining were also used or performed to validate the results. RESULTS: Expression of 4-HNE was significantly increased in both glomeruli and tubulointerstitium. At transcriptomic level, 19 FR-DEGs in glomeruli and 15 FR-DEGs in tubulointerstitium including genes of iron metabolism, antioxidant system inhibitors, and ferroptosis suppressors were significantly altered in LN. Of which, LTF, CYBB, and CCL5 were upregulated and G0S2 and AKR1C1 were downregulated in both glomeruli and tubulointerstitium of LN. qPCR further validated the alteration of LTF, CYBB, CCL5, G0S2, and AKR1C1 in the whole kidney. Correlation analysis showed that CYBB positively correlated with monocyte infiltration in glomeruli and positively correlated with response to therapy. CONCLUSION: Lipid peroxidation was aberrantly activated in LN, suggesting the activation of ferroptosis. LTF, CYBB, CCL5, G0S2, and AKR1C1, especially CYBB, might be good biomarkers of ferroptosis in LN.