基于单细胞转录组测序结果分析成纤维细胞促进口腔黏膜无瘢痕愈合的机制  

Analysis of the mechanism of fibroblast involvement in scarless healing of oral mucosa based on single cell transcriptome sequencing

作　　者：轩辕欣阳 徐佳珵 王梦婷 张立 徐慧[1] XUANYUAN Xinyang;XU Jiacheng;WANG Mengting;ZHANG Li;XU hui(Department of Dermatology,Shanghai Ninth People’s Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院皮肤科,上海市200011

出　　处：《组织工程与重建外科》2023年第2期115-123,共9页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：黄浦区产业扶持基金(XK2020007)。

摘　　要：目的探索人类正常口腔黏膜与皮肤的成纤维细胞异质性,以及口腔黏膜成纤维细胞促进伤口无瘢痕愈合的机制。方法从GEO数据库中获取13例口腔黏膜和8例皮肤的单细胞转录组测序数据。对整合后的64633个细胞进行质控、降维、聚类及细胞注释。提取成纤维细胞进行亚群定义,差异基因富集、细胞互作分析。结果从12个细胞亚群中识别出成纤维细胞,并根据基因表达特征定义为5个功能亚群:促炎型、调脂型、间充质型、收缩型和Wnt抑制型。与皮肤相比,口腔黏膜中的促炎型成纤维细胞比例显著上调,且与免疫细胞招募相关。口腔黏膜成纤维细胞高表达CXCL13、EEF1A1和B2M等基因。基因本体(GO)功能分析显示,口腔黏膜成纤维细胞中免疫应答及促炎通路等瘢痕愈合相关通路被激活。细胞互作分析显示,成纤维细胞亚群间的互作模式显著改变,口腔黏膜胶原信号通路激活的同时,包含MMP1、MMP10等基因的胶原降解基因集评分也显著上调(P<0.05)。结论正常皮肤与口腔黏膜中的成纤维细胞,其亚群、功能等存在显著差异,口腔黏膜成纤维细胞更强的促炎特性及胶原代谢水平可能是伤口无瘢痕愈合的原因。Objective To explore the heterogeneity of human normal oral mucosa and skin fibroblasts and the mechanism of oral mucosal fibroblasts in promoting scarless wound healing.Methods Single-cell transcriptome sequencing data were obtained from the GEO database for 13 cases of oral mucosa and 8 cases of skin.The 64,633 cells were subjected to quality control,dimensionality,clustering and cell annotation.Fibroblasts were extracted and annotated for subsets definition,differential gene enrichment and intercellular interactions analysis.Results Fibroblasts can be divided into 5 subsets:inflammatory,adipose,mesenchymal,contractile,and Wnt-inhibited.The proportion of inflammatory fibroblasts was significantly upregulated in the oral mucosa and was associated with immune cell recruitment.CXCL13,EEF1A1 and B2M genes were highly expressed in oral mucosa fibroblasts.Gene ontology(GO)functional analysis showed that scar healing-related pathways such as immune response and pro-inflammatory pathways were activated in oral fibroblasts.Cellular interaction analysis revealed that the interaction between fibroblast subsets was significantly changed,the information flow of oral mucosal COLLAGEN signaling pathway was upregulated,and the score of collagen degradation genes including MMP1 and MMP10 were also increased(P<0.05).Conclusion Fibroblasts in normal skin and oral mucosa differ significantly in their subpopulation and function.The stronger inflammatory characteristics and collagen metabolism of oral mucosal fibroblasts may drive the scarless wound healing.

关 键 词：皮肤 口腔黏膜 单细胞测序 成纤维细胞 无瘢痕愈合 

分 类 号：R619.6[医药卫生—外科学]