基于单细胞RNA测序探讨肾SOX9阳性祖细胞转录组特征

Study on transcriptome characteristics of renal SOX9-positive progenitor cells based on single-cell RNA sequencing

目的:基于单细胞RNA测序探究人正常肾脏中SOX9阳性祖细胞的转录组特征,为肾脏再生医学研究提供依据。方法:从GEO数据库中收集已发表的人类正常肾脏的单细胞RNA测序数据,结合差异基因分析、功能富集分析和拟时序分析探究SOX9阳性祖细胞的转录组特征,通过免疫组化实验和免疫荧光实验验证相关分析结果。结果:在人正常肾脏组织中,SOX9阳性细胞主要存在于近端小管中;GO分析结果显示,近端小管中的SOX9阳性细胞仍维持祖细胞样特性;进一步的亚群分析显示,近端小管SOX9阳性细胞可分为C1、C2和C33个亚群,其中C2亚群高表达SOX4、HES1、MYC、KLF4等与干细胞状态维持的相关基因,并在SOX9阳性细胞分化轨迹中处于起始位置。结论:人正常肾脏SOX9阳性细胞中仍有一部分细胞维持祖细胞样特性,具有潜在的再生医学研究价值。

Objective:To explore the transcriptome characteristics of SOX9-positive progenitor cells in normal human kidneys based on single-cell RNA sequencing,so as to provide a basis for renal regenerative medicine research.Methods:Published single-cell RNA sequencing data of normal human kidneys were collected from GEO database.The transcriptome characteristics analysis of SOX9-positive progenitor cells in normal human kidneys was performed by differential gene analysis,functional enrichment analysis and pseudotime trajectories analysis.The results of relevant analysis were verified by immunohistochemical and immunofluorescence experiments.Results:In our study,SOX9-positive cells were mainly found in proximal tubules in normal human kidney tissue.GO analysis showed that SOX9-positive cells in proximal tubules still maintained progenitor cell like characteristics.Further subpopulation analysis showed that the proximal tubule SOX9-positive cells could be divided into C1,C2 and C3 subgroups.Among them,C2 subgroup highly expressed SOX4,HES1,MYC,KLF4 and other genes related to stem cell state maintenance,and was in the starting position in the differentiation trajectory of SOX9-positive cells.Conclusion:Some SOX9-positive cells in human kidney still maintain progenitor cell like characteristics,which have potential value in regenerative medicine research.