ChIP-seq技术分析突变IHH-GLI1信号传导通路对下游转录调控的影响  

ChIP-seq Analysis of the Impact of Mutant IHH-GLI1 Signaling Pathway on Downstream Transcriptional Regulation

作　　者：王挺 沈陆 魏慕筠 周伟 李沫 贺林[1] 秦胜营 WANG Ting;SHEN Lu;WEI Mu-yun;ZHOU Wei;LI Mo;HE Lin;QIN Sheng-ying(Bio-X Institutes, School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200030, China)

机构地区：[1]上海交通大学生命科学技术学院Bio-X研究院,中国上海200030

出　　处：《湖南师范大学自然科学学报》2020年第6期27-35,共9页Journal of Natural Science of Hunan Normal University

基　　金：上海市2011年度“科技创新行动计划”重大科技项目子课题(11dz1950300)。

摘　　要：为发现IHH/GLI1突变导致的下游转录调控的变化,在小鼠间充质干细胞C3H10T1/2中加入人野生型重组IHH-N蛋白(WT)和突变型蛋白(MT,p.E95K)以激活IHH信号通路,利用染色体免疫沉淀高通量测序(ChIP-seq)技术挖掘GLI1转录因子结合靶点,并对其进行基因功能注释和KEGG通路分析,再结合前期基因芯片数据进行联合分析。利用ChIP-seq生物信息学分析发现两组间465个不同的GLI1结合靶点,基因注释和KEGG通路分析的结果主要富集于Wnt、刺猬因子(HH)、胰岛素等参与调控长板软骨发育的关键信号通路。联合分析发现了两个数据集中19个共同的候选基因。本研究精确定位了IHH信号蛋白突变导致的GLI1结合靶点差异,为进一步探索由IHH信号介导的BDA1的发病机制和治疗靶点提供理论基础。To explore the mechanism about the changes in regulation of downstream transcription caused by IHH/GLI1 mutation,in this work,the genome-wide binding profile of GLI1 was used for ChIP-seq analysis.C3H10T1/2 cells in the mice model were employed with wild-type recombinant IHH-N protein(WT)and mutant protein(MT,p.E 95K)to activate the IHH signaling pathway.GLI1 transcription factor binding targets were excavated by using chromosome immunoprecipitation high-throughput sequencing(ChIP-seq),and gene function annotation and KEGG pathway analysis were also performed.Meanwhile,GLI1 binding target genes analyzed by ChIP-seq were verified in microarray-based gene expression data.Our results from Chip-seq bioinformatics analysis show that there are 465 different GLI1 binding genes between the two groups.KEGG pathway enrichment analysis results of differential genes suggest that they are mainly involved in key signaling pathways that regulate long plate chondrogenesis such as Wnt,hedgehog factor(HH)and insulin.Our verification results in expression profile data indicate that there are 19 common candidate genes.Different GLI1 binding targets caused by mutations in IHH signaling proteins have been precisely located.These results chould provide a theoretical basis for the further exploration of pathogenesis and therapeutic targets of BDA1 mediated by IHH signaling.

关 键 词：A1型短指症 印度刺猬因子(IHH) GLI1 CHIP-SEQ 靶基因 

分 类 号：Q78[生物学—分子生物学] R285.5[医药卫生—中药学]