Integrated multi-omics and single-cell analysis reveals CDKN2A-mediated cuproptosis mechanisms driving thyroid carcinoma progression

Abstract Cuproptosis is a recently identified copper-dependent cell death pathway with growing relevance in tumor biology, yet its involvement in thyroid carcinoma (TC) remains poorly understood. In this study, we integrated multi-omics datasets to characterize the functional roles of cuproptosis-related genes (CRGs) in TC progression. Bulk and single-cell transcriptomic datasets from public repositories were analyzed to classify TC into two CRG-based molecular subtypes that showed significant associations with clinicopathological features and immune cell infiltration. A prognostic model derived from CRG-related differentially expressed genes exhibited high predictive accuracy for patient survival. CDKN2A emerged as the only consistently upregulated CRG in TC and correlated with adverse prognosis. Single-cell analyses further revealed distinct cellular distributions of CRGs within the tumor microenvironment, with notable enrichment in immune cell populations. In addition, a previously unrecognized competing endogenous RNA network, the GAS5/miR-128-3p/CDKN2A axis, was identified and experimentally validated. Functional assays demonstrated that this regulatory circuit modulates TC cell proliferation, invasion, and metastasis in vitro and in vivo, with GAS5 acting for miR-128-3p to regulate CDKN2A expression. These findings provide a comprehensive systems-level perspective on cuproptosis-related mechanisms in TC and highlight the therapeutic promise of targeting the cuproptosis pathway and its regulatory networks in thyroid cancer management.