CDKN1A attenuates ferroptosis in renal tubular epithelial cells and alleviates calcium oxalate crystal deposition under hyperoxaluric conditions

Abstract

Hyperoxaluria is a recognized risk factor for calcium oxalate nephrolithiasis, often contributing to tubular injury and crystal deposition. This study aimed to investigate the involvement of ferroptosis in hyperoxaluria-induced renal damage and identify key regulatory genes with therapeutic relevance. In vivo and in vitro hyperoxaluria models were established, and ferroptosis-related differentially expressed genes (FRDEGs) were screened via GEO and FerrDb databases. Functional enrichment analyses were conducted using GO and KEGG. CDKN1A was identified as a hub gene through PPI network analysis and subsequently validated in both in vivo and in vitro hyperoxaluria models via RT-qPCR and Western blotting. Concurrently, activation of ferroptosis was observed under hyperoxaluric conditions in both models, as indicated by changes in key ferroptosis markers. In HK-2 cells, CDKN1A expression decreased upon Erastin exposure, and its knockdown enhanced ferroptosis sensitivity, confirming its potential suppressive role. Additionally, analysis of the human dataset GSE73680 revealed a significant upregulation of CDKN1A expression in Randall's plaques from calcium oxalate stone patients and enrichment of ferroptosis-related pathways, consistent with findings from the mouse dataset, supporting its relevance in crystal-associated pathology. Furthermore, candidate compounds potentially activating CDKN1A were predicted based on GTEx-derived expression profiles. These findings suggest that ferroptosis contributes to hyperoxaluria-associated renal epithelial injury and crystal deposition. CDKN1A may exert a protective role by modulating ferroptosis, providing mechanistic insight into oxalate-induced kidney injury and offering a basis for developing strategies to mitigate the risk of calcium oxalate stone formation.

Keywords: CDKN1A; Calcium oxalate crystal deposition; Ferroptosis; Hyperoxaluria.