结核表位疫苗MP3RT在人源化小鼠模型上的转录组学分析  

Transcriptomic analysis of tuberculosis peptide-based vaccine MP3RT in humanized mice

作　　者：王杰 米洁 梁艳[1] 吴雪琼[1] 张俊仙[1] 刘银萍[1] 王兰[1] 薛勇 史迎昌[1] 龚文平 Wang Jie;Mi Jie;Liang Yan;Wu Xueqiong;Zhang Junxian;Liu Yinping;Wang Lan;Xue Yong;Shi Yingchang;Gong Wenping(Tuberculosis Prevention and Control Key Laboratory/Beijing Key Laboratory of New Techniques of Tuberculosis Diagnosis and Treatment,Senior Department of Tuberculosis,the Eighth Medical Center of PLA General Hospital,Beijing 100091,China)

机构地区：[1]解放军总医院第八医学中心结核病医学部研究所军队结核病防控重点实验室/结核病诊疗新技术北京市重点实验室,北京100091

出　　处：《中华结核和呼吸杂志》2022年第9期894-903,共10页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金(81801643);北京市自然科学基金(19L2065);解放军总医院青年基金(QNC19047)。

摘　　要：目的通过转录组学确定结核表位疫苗MP3RT在人源化动物模型上诱导的差异表达基因(DEG),为阐明MP3RT潜在的分子机制提供新思路。方法本研究于2019年8月开始至2022年2月完成。本研究采用edgeR软件以差异倍数≥1.5且P<0.05为筛选条件筛DEG,对筛选出的DEG进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)分析以及蛋白互作网络分析。并对上述DEG通过RT-qPCR进行实验验证,采用GraphPad Prism 8软件进行统计学分析。结果转录组学鉴定了367个DEG(214个上调,153个下调)。生物信息学分析发现,上述DEG的GO富集显著聚焦于细胞代谢、生长、凋亡、炎症等词条,KEGG富集显著聚焦于MAPK信号通路等炎症相关通路。蛋白互作网络分析显示,蛋白Abl1聚集度最大,聚集系数最高,连通性最好。RT-qPCR结果显示,与磷酸盐缓冲液(PBS)组相比,MP3RT疫苗组中cpne4(t=2.48,P=0.0480)、h2-q10(t=2.95,P=0.0256)、mef2c(t=2.87,P=0.0284)、cr2(t=3.23,P=0.178)、ablim1(t=2.91,P=0.0335)、dll1(t=2.70,P=0.0273)和ms4a2(t=3.03,P=0.0192)基因表达水平显著上调,cd163l1(t=2.56,P=0.0430)、il1r1(t=2.91,P=0.0227)和cd34(t=2.42,P=0.0462)基因表达水平显著下调。结论MP3RT表位疫苗在人源化动物模型上诱导了367个DEG,这些DEG与新陈代谢和免疫反应相关,p38 MAPK和JNK/MAPK信号通路在MP3RT疫苗分子机制中扮演着重要的角色。Objective To identify the differentially expressed genes(DEGs)induced by tuberculosis peptide-based vaccine MP3RT in a humanized mouse model using transcriptomics technology.Methods This study was conducted from August 2019 to February 2022.We used edgeR software to screen DEGs with a fold change greater than or equal to 1.5 and a P value less than 0.05 as screening conditions.Gene ontology(GO),Kyoto encyclopedia of genes and genomes(KEGG),and protein interaction network analyses were performed on the screened DEGs.Then,these DEGs were verified by RT-qPCR and statistically analyzed by GraphPad Prism 8 software.Results A total of 367 DEGs(214 up-regulated and 153 down-regulated)were identified by transcriptomics.Bioinformatics analysis showed that the GO enrichment of the DEGs mentioned above significantly focused on cell metabolism,growth,apoptosis,inflammation,and other terms.In contrast,the KEGG enrichment significantly focused on inflammatory pathways such as the MAPK signaling pathway.Protein interaction network analysis showed that protein Abl1 had the highest aggregation,the highest aggregation coefficient,and the best connectivity.RT-qPCR results showed that gene expressions of cpne4(t=2.48,P=0.0480),h2-q10(t=2.95,P=0.0256),mef2c(t=2.87,P=0.0284),cr2(t=3.23,P=0.178),ablim1(t=2.91,P=0.0335),dll1(t=2.70,P=0.0273)and ms4a2(t=3.03,P=0.0192)genes in the MP3RT group were significantly up-regulated than those in the PBS group,while gene expressions of cd163l1(t=2.56,P=0.0430),il1r1(t=2.91,P=0.0227)and cd34(t=2.42,P=0.0462)genes in the MP3RT group were significantly down-regulated than those in the PBS group.Conclusions The MP3RT vaccine induced 367 DEGs in humanized mice,which were associated with metabolic and immune responses.Furthermore,we found that p38 MAPK and JNK/MAPK signaling pathways played an important role in the molecular mechanism of the MP3RT vaccine.

关 键 词：结核分枝杆菌 表位疫苗 MP3RT 转录组学 差异表达基因 

分 类 号：R392-33[医药卫生—免疫学]