Glycolysis-related gene signatures and the functional role of P4HA1 in osteosarcoma prognosis

Osteosarcoma, a primary malignant bone tumor predominantly affecting children and adolescents, is characterized by aerobic glycolysis, which is intricately linked to tumor progression and metastasis, yet its prognostic implications remain underexplored. This study aimed to develop a prognostic model utilizing glycolysis-related genes and to elucidate the functional role of P4HA1, a key gene within this model, in osteosarcoma prognosis and immune cell infiltration. We collected clinical and transcriptomic data from osteosarcoma patients in the UCSC Xena and GEO databases. Through univariate Cox and LASSO regression analyses, we identified 12 glycolysis-related genes that significantly influence osteosarcoma prognosis. These genes were employed to construct a risk score model, which accurately predicted patient outcomes as demonstrated by survival analysis and ROC curves, with an AUC of 0.899, 0.881, and 0.878 for 1-year, 3-year, and 5-year survival predictions, respectively. The model was particularly effective across different clinical subgroups. Immune cell infiltration analysis revealed that CD8⁺ T cells, naïve CD4⁺ T cells, resting dendritic cells, and activated mast cells significantly contributed to the model's predictive power. The model also showed significant enrichment of immune-related signaling pathways, indicating a robust association between immune status and glycolytic-related risk scores in osteosarcoma prognosis. Notably, P4HA1 was upregulated in osteosarcoma tissues and promoted cell proliferation in a glycolysis-dependent manner, as evidenced by increased intracellular ATP levels, inhibited glucose absorption, and elevated lactate levels in P4HA1-overexpressing osteosarcoma cells. The promotion of proliferation by P4HA1 could be significantly attenuated by the glycolysis inhibitor 2-DG, highlighting the glycolysis dependency of P4HA1's action. In conclusion, we developed a prognostic model for osteosarcoma by integrating glycolysis-related genes, with a particular emphasis on the functional role of P4HA1. Our findings highlight the interplay between glycolysis and immune cell infiltration in disease prognosis. This model provides insights for targeted therapies and a foundation for further research into osteosarcoma treatment.