Integrative oxidative stress–disulfidptosis analysis identifies NPR3 and ZNF366 as cell-type-associated candidate markers in clear cell renal cell carcinoma

Disulfidptosis is a regulated cell-death program that occurs in SLC7A11-high cells under glucose limitation, but its relationship with oxidative stress in clear cell renal cell carcinoma (ccRCC) remains unclear. We integrated 313 oxidative stress-related genes with 24 disulfidptosis-related genes in the original TCGA-KIRC analysis. Pearson screening (|r| ≥ 0.8 and nominal two-sided P < 0.05) identified 21 disulfidptosis-oxidative stress (DOS) genes, from which three reported DOS subtypes, 63 subtype-associated genes and a four-gene score comprising ZNF366 , EMCN , GRIK3 and NPR3 were derived. In an updated GDC-derived TCGA-KIRC dataset, both k = 3 classifications showed method-dependent stability, with weaker stability for the available 58-of-63-gene solution. Outcome-blind centroid transfer assigned all 101 E-MTAB-1980 cases and reproduced several molecular and immune/stromal differences in GSE73731, although the hard clusters have not been independently validated for prognosis. In the atezolizumab-containing arms of IMmotion150, the within-cohort re-standardized four-gene score was lower in patients with clinical benefit and was associated with progression-free survival; the DOS21 module score was not associated with clinical benefit. The fixed four-gene score had modest internal discrimination (bootstrap optimism-corrected C-index, 0.697), and its transfer was sensitive to platform scaling and stage adjustment. Under the single 6-h glucose-deprivation condition, NPR3 and ZNF366 increased only in SLC7A11-high UMRC6 cells. Single-cell analyses in one VHL-related case and an independent 19-patient ccRCC cohort supported endothelial enrichment of ZNF366 and showed that NPR3 was not restricted to tumor cells. These findings identify NPR3 and ZNF366 as compartment-associated candidate markers but do not establish a functional role in disulfidptosis.