Identification and validation of autophagy-related genes during osteogenic differentiation of bone marrow mesenchymal stem cells

Objective(s): Osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is an essential stage in bone formation. Autophagy plays a pivotal role in the self-renewal potential and pluripotency of stem cells. This study aimed to explore the function of autophagy-related genes during osteogenic differentiation of BMSCs.Materials and Methods: The differentially expressed autophagy-related genes (ARGs) were obtained from the GEO and HADb databases. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed using R software. The PPI and hub gene mining networks were constructed using the STRING database and Cytoscape. Finally, the RT-qPCR was conducted to validate the expression level of ARGs in BMSCs.Results: Thirty-seven differentially expressed ARGs were finally obtained, including ۱۲ upregulated and ۲۵ downregulated genes. GO and KEGG enrichment analysis showed that most of these genes were enriched in apoptosis and autophagy. The PPI network revealed strong interactions between differentially expressed ARGs. The expression level of differentially expressed ARGs tested by RT-qPCR showed ۶ upregulated ARGs, including FOXO۱, MAP۱LC۳C, CTSB, FOXO۳, CALCOCO۲, FKBP۱A, and ۴ downregulated  ARGs, including MAPK۸IP۱, NRG۱, VEGFA, and ITGA۶ were consistent with the expression of high-throughput sequencing data.Conclusion: We identified ۳۷ ARGs during osteogenic differentiation using bioinformatics analysis. FOXO۱, MAP۱LC۳C, CTSB, FOXO۳, CALCOCO۲, FKBP۱A, MAPK۸IP۱, NRG۱, VEGFA, and ITGA۶ may regulate osteogenic differentiation of hBMSCs by involving autophagy pathway. This study provides new insight into the osteogenic differentiation of hBMSCs and may be available in developing therapeutic strategies for maxillofacial bone defects.

Keywords:

Autophagy , Bioinformatics , Bone marrow mesenchymal stem cells , bone regeneration , Osteogenesis