单细胞测序分析人类胚胎干细胞神经分化的分子机制  

Molecular mechanism of neural differentiation in human embryonic stem cells by single-cell sequencing analysis

作　　者：曹冰 张晓明[2] 梁富龙 Cao Bing;Zhang Xiaoming;Liang Fulong(Department ofMedical Director,Xiamen Fifth Hospital,Xiamen 361000,China;Stroke Center the Second Affiliated Hospital of Harbin Medical University,Harbin 150000,China;Department of Neurology,Xiamen Fifth Hospital,Xiamen 361000,China)

机构地区：[1]厦门市第五医院医务总监科,361000 [2]哈尔滨医科大学附属第二医院卒中中心,150000 [3]厦门市第五医院神经内科,361000

出　　处：《中华细胞与干细胞杂志（电子版）》2021年第1期1-7,共7页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：厦门市第五医院科研课题(50193)。

摘　　要：目的探讨人类胚胎干细胞 (ESCs)分化为神经细胞的关键性靶基因及分子机制,为临床靶向治疗神经康复患者提供分子理论依据。方法基于GEO数据平台芯片,采用单细胞测序方法 (scRNA-seq),利用R语言从多分子维度(单细胞差异基因、蛋白互作网络和基因通路等)分析人类ESCs分化过程中的关键Marker基因并利用质控和数据过滤、PCA、TSNE分析、细胞轨迹分析、GO富集分析、KEGG富集分析、KEGG通路分析等论证Marker基因调控人类ESCs分化作用机制。结果 GO功能富集分析结果为:Marker基因在胚层分化、细胞外基质和信号转导中作用显著;Marker基因互作网络及KEGG通路显示了特征性纤维连接蛋白1(FN1)、同源域蛋白 (NANOG)、生长因子受体结合蛋白2 (GRB2)为关键基因;KEGG通路分析显示:(1)FN1在调控细胞外基质通路中作用显著;(2)NANOG、GRB2在调控干细胞多能性信号通路作用显著。结论 FN1可能通过调控细胞外基质通路介导人类ESCs基质环境的变化;NANOG、GRB2上调在干细胞多能性信号通路中高表达介导人类ESCs定向分化为神经组织。Objective To explore the key target genes and molecular mechanisms of human embryonic stem cell differentiation into neural cells,and provide molecular theoretical basis for clinical targeted treatment of neurological rehabilitation patients.Methods Based on the chip data from GEO platform,the key Marker genes in the differentiation process of human embryonic stem cells were analyzed from the multi-molecular dimension(single cell differential genes,protein interaction networks,gene pathways,etc.)by using R language as well as using single cell sequencing method.Quality control and data filtering,PCA,TSNE analysis,cell trajectory analysis,GO enrichment analysis,KEGG enrichment analysis,KEGG pathway analysis were used to show the mechanism of Marker gene regulating human embryonic stem cell differentiation.Results GO functional enrichment analysis showed that Marker genes played a significant role in germ layer differentiation,extracellular matrix and signal transduction.Marker gene interaction network and KEGG pathway showed that characteristic Fibronectin 1(FN1),Nanog homeobox(NANOG)and Growth factor receptor-bound protein 2(GRB2)genes were the key genes.KEGG pathway analysis showed that FN1 played a significant role in regulating extracellular matrix pathway,and NANOG and GRB2 played significant roles in regulating stem cell pluripotency signaling pathway.Conclusion FN1 may mediate the matrix environment of human embryonic stem cells by regulating extracellular matrix pathways and up-regulation of NANOG and GRB2 in stem cell pluripotency signaling pathway could mediate directional differentiation of human embryonic stem cells into neural tissue.

关 键 词：人类胚胎干细胞 神经祖细胞 单细胞测序 靶基因 分子机制 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]