Integrative multi-omics identifies PANX2 as a ferroptosis suppressor and potential therapeutic target in clear cell renal cell carcinoma
Background Clear cell renal cell carcinoma (ccRCC) is metabolically primed for ferroptosis, yet the prognostic relevance and mechanistic contribution of ferroptosis-related genes remain incompletely defined. This study aimed to identify ferroptosis-associated biomarkers with prognostic value and to clarify their functional relevance in ccRCC progression. Methods We integrated single-cell RNA sequencing, bulk RNA-seq, spatial transcriptomics, and machine-learning-based feature selection to identify ferroptosis-related prognostic genes in ccRCC. A four-gene risk model and an integrated nomogram were constructed and evaluated in independent cohorts. PANX2 was prioritized for experimental validation using stable knockdown models, RNA sequencing, lipid peroxidation and iron probes, redox assays, Western blot, luciferase reporter assays, xenografts, and an immunocompetent murine renal carcinoma model. Results A four-gene prognostic signature (CA9, PVT1, RRM2, and PANX2) was identified and used to construct a risk model with consistent predictive performance in the training and external validation cohorts. Among these genes, PANX2 was predominantly enriched in epithelial tumor compartments and had not been functionally characterized in ccRCC. PANX2 knockdown inhibited ccRCC cell proliferation, reduced antioxidant capacity, increased intracellular Fe2+ accumulation and lipid peroxidation, and sensitized cells to erastin-induced ferroptotic death. Mechanistically, PANX2 loss was associated with reduced Akt/mTOR pathway activity and diminished SLC7A11 expression; rescue with an Akt activator or SLC7A11 overexpression attenuated ferroptosis-associated phenotypes. In an immunocompetent murine renal carcinoma model, PANX2 knockdown was accompanied by increased infiltration of CD45+ leukocytes, CD3+ T cells, and CD8+ T cells, supporting a potential link between PANX2-dependent ferroptosis resistance and the tumor immune contexture. Conclusions This study identifies PANX2 as a ccRCC-relevant suppressor of ferroptosis and supports the involvement of a PANX2-Akt/mTOR-SLC7A11-associated signaling axis in redox homeostasis and tumor progression. The ferroptosis-related prognostic model and nomogram may support risk stratification, while PANX2 represents a candidate therapeutic vulnerability that warrants further mechanistic and translational validation.