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[人源化小鼠中基于结核肽的疫苗MP3RT的转录组学分析]

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

作者信息

Wang J, Mi J, Liang Y, Wu X Q, Zhang J X, Liu Y P, Wang L, Xue Y, Shi Y C, Gong W P

机构信息

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.

出版信息

Zhonghua Jie He He Hu Xi Za Zhi. 2022 Sep 12;45(9):894-903. doi: 10.3760/cma.j.cn112147-20220112-00045.

DOI:10.3760/cma.j.cn112147-20220112-00045
PMID:36097927
Abstract

To identify the differentially expressed genes (DEGs) induced by tuberculosis peptide-based vaccine MP3RT in a humanized mouse model using transcriptomics technology. 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 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. 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 (, (=2.95, =0.025 6), (=2.87, =0.028 4), (=3.23, =0.178), (=2.91, =0.033 5), (=2.70, =0.027 3) and (=3.03, =0.019 2) genes in the MP3RT group were significantly up-regulated than those in the PBS group, while gene expressions of (=2.56, =0.043 0), (=2.91, =0.022 7) and (=2.42, =0.046 2) genes in the MP3RT group were significantly down-regulated than those in the PBS group. 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.

摘要

利用转录组学技术,在人源化小鼠模型中鉴定基于结核肽的疫苗MP3RT诱导的差异表达基因(DEGs)。本研究于2019年8月至2022年2月进行。我们使用edgeR软件,以倍数变化大于或等于1.5且P值小于0.05作为筛选条件来筛选DEGs。对筛选出的DEGs进行基因本体论(GO)、京都基因与基因组百科全书(KEGG)以及蛋白质相互作用网络分析。然后,通过RT-qPCR对这些DEGs进行验证,并使用GraphPad Prism 8软件进行统计分析。通过转录组学共鉴定出367个DEGs(214个上调和153个下调)。生物信息学分析表明,上述DEGs的GO富集显著集中在细胞代谢、生长、凋亡、炎症等方面。相比之下,KEGG富集显著集中在如MAPK信号通路等炎症途径。蛋白质相互作用网络分析表明,蛋白质Abl1具有最高的聚集度、最高的聚集系数和最佳的连通性。RT-qPCR结果显示,MP3RT组中(,(F = 2.95,P = 0.025 6),(F = 2.87,P = 0.028 4),(F = 3.23,P = 0.178),(F = 2.91,P = 0.033 5),(F = 2.70,P = 0.027 3)和(F = 3.03,P = 0.019 2)基因的表达明显高于PBS组,而MP3RT组中(F = 2.56,P = 0.043 0),(F = 2.91,P = 0.022 7)和(F = 2.42,P = 0.046 2)基因的表达明显低于PBS组。MP3RT疫苗在人源化小鼠中诱导了367个DEGs,这些DEGs与代谢和免疫反应相关。此外,我们发现p38 MAPK和JNK/MAPK信号通路在MP3RT疫苗的分子机制中发挥了重要作用。

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