口腔疾病防治2026，Vol.34Issue(7)：642-656,15.DOI:10.12016/j.issn.2096-1456.202660042
单细胞组学解析ACVR1在成牙本质细胞分化中的作用
Single-cell omics reveals the role of ACVR1 in odontoblastic differentiation
摘要
Abstract
Objective To investigate the impact of activin receptor type-1(ACVR1)on odontoblastic differentiation and dentin formation based on single-cell RNA sequencing.Methods A correlation analysis of ACVR1 with odontogenesis-related transcription factors was performed using single-cell data from healthy wild-type adult mouse inci-sors obtained from the Gene Expression Omnibus database(GSE146123).This experiment was approved by the Institu-tional Animal Care and Use Committee of the affiliated institution.Conditional knockout mice with ACVR1 deleted in the dental mesenchyme were generated using the Cre-LoxP system(experimental group:Osterix-Cre;ACVR1fx/-,control group:Osterix-Cre;ACVR1fx/+).Dental pulp tissue was extracted from the incisors of 3-week-old mice to prepare single-cell suspensions for 10×Genomics single-cell transcriptome sequencing.Quality control,normalization,dimensionality reduction,and Harmony batch effect correction were performed using the Seurat pipeline,followed by UMAP dimen-sionality reduction visualization and cell subpopulation identification.Monocle was used for pseudotime analysis to infer cell differentiation trajectories,and changes in transcription factors positively correlated with ACVR1 were ana-lyzed at the single-cell data level.Immunofluorescence staining was employed to validate the localization and expres-sion changes of the key transcription factor Osterix and the odontoblast-specific protein DSPP.Results Sp7/Osterix is widely expressed in the incisor mesenchyme of mice(GSE146123).Using Osterix-Cre,ACVR1 was effectively knocked out in the mesenchyme.Gene correlation analysis between ACVR1 and key odontogenic genes showed positive correla-tions between ACVR1 and genes such as Msx1,Msx2,and Sp7.Single-cell atlases of incisor dental pulp tissue from"Osterix-Cre;ACVR1fx/-"and"Osterix-Cre;ACVR1fx/+"mice were then constructed.The most significant increase in the proportion of pre-odontoblasts was in the experimental group,along with downregulated expression of key genes re-lated to odontoblastic differentiation and polarization in this subpopulation.Differential gene enrichment analysis across all mesenchymal subpopulations indicated downregulated expression of most genes associated with dentin formation.Fi-nally,as observed through single-cell differential gene analysis,pseudotime expression analysis,and immunofluores-cence validation,the expression of Sp7 and DSPP were significantly downregulated in the experimental group.Conclu-sion The ACVR1 gene may promote odontoblastic differentiation and subsequently affect dentin formation through transcription factors such as Sp7,Msx1,and Msx2.关键词
成牙本质细胞分化/活化蛋白A受体1型/Msx1转录因子/Msx2转录因子/Sp7转录因子/牙源性间充质细胞/单细胞测序/生物信息学分析Key words
odontoblastic differentiation/activin receptor type-1/Msx1 transcription factor/Msx2 transcrip-tion factor/Sp7 transcription factor/dental mesenchymal cells/single-cell sequencing/bioinformatics analysis分类
医药卫生引用本文复制引用
宋少艺,刘苍维,史册,刘麒麟,孙宏晨..单细胞组学解析ACVR1在成牙本质细胞分化中的作用[J].口腔疾病防治,2026,34(7):642-656,15.基金项目
国家自然科学基金项目(82370936) This study was supported by the grant from National Natural Science Foundation of China(No.82370936).The funder had no role in the study design,data collection and analysis,decision to publish,or preparation of the manuscript. （82370936）
