Analysis of key transcription factors and cellular communication in osteoblast development

LIU Yu'an; LIU Yan; YANG Liu; ZHENG Chao

doi:10.13276/j.issn.2097-1656.2024.11.016

Abstract：Objective To analyze key transcription factors and cellular communication for developmental regulation using osteoblast single-cell sequencing data.Methods The osteoblast single-cell sequencing dataset from GEO database was subjected to dimensionality reduction,clustering,and annotation,and the annotated subpopulations of osteoblasts were analyzed for transcription factor regulatory networks and regulatory activities using pySCENIC software package,and the binary regulator activity scores were obtained using the binarization function binarize to identify the characteristic transcription factors of osteoblasts at different stages of maturation and the driving transcription factors at different stages of differentiation,respectively.Ligand-receptor interactions and cell-cell communication networks were predicted from single-cell transcriptome data using CellCall,and potential interaction strengths and key signaling pathways between osteoblast subpopulations at different stages of maturation were predicted based on the expression of ligand-receptor pairs and transcription factors.Results The C6 subpopulation of terminally differentiated osteoblasts communicated with the C3 subpopulation through SOST/DKK1 paracrine secretion,which in turn led to the inhibition of Wnt signaling and the reduction of Tcf7l2 transcriptional activity in the C3 subpopulation.At the same time,Notch signaling pathway and downstream transcription factor Rbpj were activated through cellular contact between C3-C4 and C4-C4,which in turn finely regulated the maturation and differentiation of osteoblasts to osteocytes.Conclusion Osteoblast-to-osteocyte differentiation is regulated by the ligand-receptor-transcription factor-mediated Wnt and Notch signaling pathways.