Identification of hub genes in monocyte macrophages of high and low peak bone mass populations and early diagnostic biomarkers for osteoporosis

Osteoporosis (OP) , a degenerative condition defined by osteopenia, is strongly influenced by peak bone mass (PBM) . However, the early diagnosis of osteoporosis remains incompletely understood. This study aims to identify early diagnostic biomarkers in populations with low PBM and to validate their clinical significance in the diagnosis, treatment, and prevention of osteoporosis. We obtained three microarray datasets (GSE2208, GSE97498, and GSE64433) from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were screened using the limma package. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed. Protein-protein interaction network (PPI) analysis and visualization were conducted with String and Cytoscape. CytoHubba was used to identify hub genes, and relevant miRNAs were predicted using CyTargetLinker in Cytoscape. Finally, the hub genes and predicted miRNA expression were confirmed via RT-qPCR experiments in peripheral blood monocytes of ovariectomy (OVX) mice. We identified 747 DEGs from GSE2208 and 1238 DEGs from GSE97498 and identified 58 overlapping DGEs between these two datasets. The enriched GO terms and pathways were determined, including “TNF signaling pathway, plasma membrane, protease binding, and positive regulation of I-kB kinase/NF-kB signaling.” Ten hub genes (TNF, FN-1, CCR2, HB-EGF, MMP14, NOD2, SOCS3, IFNAR1, IRAK3, PRKACB) were selected from the overlapping DEGs. Additionally, 42 target miRNAs were identified for the ten hub genes using CyTargetLinker. Eight miRNAs were selected after cross-validation with the osteoporosis miRNA expression profiling dataset (GSE64433) . In RT-qPCR experiments, the ten hub genes and eight predicted miRNAs in the blood monocytes of OVX mice were further validated. The study found that the hub genes and predicted miRNAs are potentially linked to OP development and may serve as biomarkers for the early screening of individuals at high risk of OP, thus playing a pivotal role in OP prevention and treatment. This provides a valuable foundation for further experimental studies and clinical applications.