成骨细胞发育中的关键转录因子及细胞通讯分析  

Analysis of key transcription factors and cellular communication in osteoblast development

作　　者：刘遇安 刘妍 杨柳 郑超 LIU Yu'an;LIU Yan;YANG Liu;ZHENG Chao(Cadet Brigade,School of Basic Medical Sciences,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China;Department of Ophthalmology,Eye Institute of Chinese PLA,Xijing Hospital,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China;Department of Orthopedics,Institute of Orthopedics of PLA,Xijing Hospital,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China;Department of Medical Genetics and Developmental Biology,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China)

机构地区：[1]空军军医大学基础医学院学员队,陕西西安710032 [2]空军军医大学全军眼科研究所,西京医院眼科,陕西西安710032 [3]空军军医大学全军骨科研究所,西京医院骨科,陕西西安710032 [4]空军军医大学基础医学院医学遗传与发育生物学教研室,陕西西安710032

出　　处：《空军军医大学学报》2024年第11期1278-1283,共6页Journal of Air Force Medical University

基　　金：国家自然科学基金面上项目(82372361);空军军医大学西京医院学科助推计划项目(XJZT240N54,XJZT21CM12,XJZT24QN51)。

摘　　要：目的使用成骨细胞单细胞测序数据分析发育调控关键转录因子和细胞通讯。方法对GEO数据库中成骨细胞单细胞测序数据集进行降维、聚类、注释,对注释好的成骨细胞亚群使用pySCENIC软件包,分析转录因子调控网络和调控活性,使用二值化函数binarize获得二元调控子活性评分,分别鉴定成骨细胞不同成熟阶段的特征性转录因子和不同分化阶段的驱动转录因子;使用CellCall从单细胞转录组数据中预测配体受体相互作用和细胞细胞通信网络,根据配体受体对及转录因子的表达,预测不同成熟分化阶段的成骨细胞亚群之间的潜在相互作用强度和关键信号通路。结果终末分化的骨细胞C6亚群通过SOST/DKK1旁分泌作用于C3亚群,进而使得C3亚群Wnt信号受到抑制、Tcf7l2转录活性降低;同时,C3-C4、C4-C4间通过细胞接触激活Notch信号通路、激活下游转录因子Rbpj,进而对成骨细胞向骨细胞的成熟分化进行精细的调节。结论成骨细胞向骨细胞发育过程受到配受体转录因子介导的Wnt和Notch信号通路调控。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.

关 键 词：成骨细胞 骨细胞 单细胞测序 转录因子 细胞通讯 

分 类 号：R394[医药卫生—医学遗传学]