Paraptosis landscape reveals distinct prognoses and immune microenvironments in osteosarcoma: Experimental validation...

Osteosarcoma (OS) is a highly aggressive bone cancer with complex molecular mechanisms and variable patient outcomes. This study delineates the paraptosis-related genes (PRGs) landscape in OS and its association with prognosis and the immune microenvironment using bioinformatics approaches. Sixty PRGs were identified and annotated in the Therapeutically Applicable Research to Generate Effective Treatments (TARGET) and GSE21257 cohorts. Consensus clustering stratified patients into molecular clusters, revealing significant survival differences. A risk score was developed using LASSO-Cox regression, which effectively stratified patients into high and low-risk groups. Immune profiling revealed significant differences in immune cell infiltration, with high-risk groups exhibiting decreased levels of activated immune cells. Pathway enrichment analysis indicated higher activity in WNT/BETA-CATENIN, TGF-BETA, and KRAS signaling pathways in high-risk groups. Single-cell RNA sequencing highlighted elevated TNK2 expression in malignant cells. Functional assays confirmed TNK2's role in cell proliferation, viability, and migration, and showed that TNK2 knockdown mitigated cycloheximide-induced proliferation inhibition, linking TNK2 to paraptosis regulation. These findings identify key PRGs and their prognostic significance in OS, revealing distinct immune microenvironments and signaling pathways between molecular clusters. This study also underscores the importance of TNK2 in OS pathogenesis and its potential utility in developing personalized treatment strategies.