Targeting dysregulated glycolysis in type 2 diabetic osteoporosis: Identification of a diagnostic gene signature and therapeutic validation of calcifediol.

Type 2 diabetic osteoporosis (T2DOP) is a significant complication of type 2 diabetes, characterized by an increased risk of fractures. While dysregulated bone cell glucose metabolism is implicated, its specific mechanisms are unclear. This research combined bioinformatics with experimental validation to examine glycolysis-related genes in T2DOP. We merged osteoporosis and type 2 diabetes datasets, then applied differential expression analysis, WGCNA, and machine learning (113 algorithms) to identify core biomarkers. Functional enrichment, immune infiltration, and scRNA-seq analyses elucidated their roles. L1000FWD screened potential drugs, validated by molecular docking and in a T2D mouse model. We identified eight key biomarkers (ATN1, PDLIM7, PLD3, ACADSB, PDZK1, RAPGEF4, MYBPC3, BSG) with high diagnostic value (AUC > 0.75). They were enriched in glucose metabolism and osteoclast pathways. Immune analysis revealed altered infiltration, and scRNA-seq indicated their primary influence on osteoblasts, mesenchymal stem cells, and osteoclasts. Calcifediol (25(OH)D₃) was the top drug candidate, showing strong binding affinity. In vivo, 25(OH)D₃ treatment ameliorated bone loss, improved metabolic parameters, and modulated key biomarker expression. Our research identifies eight glycolysis-related genes as candidate diagnostic and therapeutic targets with biological and causal plausibility for T2DOP. Calcifediol demonstrates promise as a preclinical therapeutic candidate for T2DOP, with the potential to enable simultaneous management of diabetic hyperglycemia and osteoporosis, and its clinical value requires further verification in subsequent studies.