Targeting TYROBP to influence the immune microenvironment and osteogenic differentiation of mesenchymal stem cells

BACKGROUND: Lactate, as an end product of glycolysis, plays an important role in cellular metabolism and signal transduction, and recent studies have shown that it is closely related to cellular differentiation, but its potential role in osteogenic differentiation has not yet been fully investigated. METHODS: We obtained two datasets containing human mesenchymal stem cells and human osteoblasts, GSE12266 and GSE18043, from the GEO database, which contained a total of 14 samples with sequencing data, and searched for lactate metabolism-related genes from the Genecards database. Ten differentially expressed core genes related to lactate metabolism were identified by differential expression analysis, protein interaction network analysis, and correlation expression analysis, and determined to play a key role in osteogenic differentiation. The effects of hub genes on the immune microenvironment of osteogenic differentiation were explored by enrichment analysis and immune infiltration analysis, and the significant effects of the key gene TYRO Protein Tyrosine Kinase-Binding Protein(TYROBP) on the characterization of bone marrow mesenchymal stem cells (BMSCs) were experimentally verified, and it was determined by drug sensitivity analysis that TYROBP may be a regulatory target of certain drugs affecting osteogenic differentiation. RESULT: We successfully screened 10 differentially expressed hub genes related to lactate metabolism, and their area under the curve AUC values for predicting osteogenic differentiation were all highly favorable. Enrichment analysis showed that lactate metabolism may affect osteoblast differentiation through immune infiltration, and the immune infiltration results confirmed the strong association between hub genes and osteoblast immune infiltration status. It was verified that decreasing TYROBP expression promoted cell viability, proliferation and migration ability of BMSCs. Drug sensitivity analysis showed that TYROBP may be a major regulator of drug-induced MSC differentiation. CONCLUSION: Our study reveals the critical role of lactate metabolism in osteoblast differentiation, identifies the role of the key gene TYROBP in the regulation of BMSCs, and provides new insights for studies related to the regulation of osteoblast differentiation.