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基于高通量测序的非动脉炎性前部缺血性视神经病变竞争性内源性RNA调控网络的构建及功能富集分析

Construction and functional enrichment analysis of the competitive endogenous RNA regulatory network for nonarteritic anterior ischemic optic neuropathy based on high-throughput sequencing.

作者信息

Zhang Meng, Wang Xinling

机构信息

Department of Ophthalmology, the Fourth Affiliated Hospital of China Medical University, Eye Hospital of China Medical University, Key Lens Research Laboratory of Liaoning Province, Shenyang City, People's Republic of China.

出版信息

Funct Integr Genomics. 2022 Dec;22(6):1253-1267. doi: 10.1007/s10142-022-00914-z. Epub 2022 Nov 10.

DOI:10.1007/s10142-022-00914-z
PMID:36355332
Abstract

Based on the transcriptome high-throughput sequencing of nonarteritic anterior ischemic optic neuropathy (NAION), this study constructed a competitive endogenous RNA (ceRNA) network, enriched and analyzed it, screened long noncoding (lnc)RNAs and circular (circ)RNAs that may participate in the competitive endogenous mechanism in NAION, and inferred its function. Four milliliters of peripheral blood from NAION patients and the control group was extracted from clinical samples, transcriptome high-throughput sequencing was performed, and the sequencing data were visualized. Based on the principle of the ceRNA network, the lncRNA-micro (mi)RNA-messenger (m)RNA interaction axis and circRNA-miRNA‒mRNA interaction axes were constructed. The differentially expressed genes (DEGs) in the interaction axis were enriched and analyzed by Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG), and the functions and signalling pathways of lncRNAs and circRNAs in the interaction network were speculated. Fifty-one circRNAs were differentially expressed in the sequencing data: 25 were upregulated, and 26 were downregulated. For 996 differentially expressed lncRNAs, 317 were upregulated and 679 were downregulated, and for 1161 differentially expressed mRNAs, 698 were upregulated and 463 were downregulated. Thirty-three differentially expressed miRNAs, upregulated miRNA 18 and downregulated miRNA 15 were identified. After screening, 13 coexpressed mRNAs, 15 lncRNAs, and 3 miRNAs were finally constructed in the lncRNA-miRNA-mRNA network, and the circRNA-miRNA-mRNA network was constructed by 159 mRNAs, 26 miRNAs, and 34 circRNAs. In the lncRNA network, GO enrichment analysis obtained 182 biological processes, 12 cell components and 38 molecular functions, which are related mainly to the regulation of the activity of proteins and enzymes such as cyclic nucleotide-dependent protein kinase activity and magnesium ion-dependent protein serine/threonine phosphatase activity. KEGG analysis involved mainly the forkhead box O (FoxO) signalling pathway, apelin signalling pathway, etc. In the circRNA enrichment results, 353 biological processes, 52 cell components, and 45 molecular functions were obtained, involving mainly calcium channel regulation, neutrophil activation, mRNA transport, and metabolism. KEGG mainly involved the Wnt signalling pathway, apelin signalling pathway, Hippo signalling pathway, oxytocin signalling pathway, etc. This paper provides a new idea for lncRNAs and circRNAs to mediate the occurrence and development of NAION through the ceRNA mechanism. This study lays a foundation for the in-depth study of the pathogenesis of NAION.

摘要

基于非动脉炎性前部缺血性视神经病变(NAION)的转录组高通量测序,本研究构建了竞争性内源RNA(ceRNA)网络,对其进行富集和分析,筛选出可能参与NAION竞争性内源机制的长链非编码(lnc)RNA和环状(circ)RNA,并推断其功能。从临床样本中提取NAION患者和对照组的4毫升外周血,进行转录组高通量测序,并将测序数据可视化。基于ceRNA网络原理,构建lncRNA-微小(mi)RNA-信使(m)RNA相互作用轴和circRNA-miRNA-mRNA相互作用轴。通过基因本体论(GO)和京都基因与基因组百科全书(KEGG)对相互作用轴中的差异表达基因(DEG)进行富集和分析,推测lncRNA和circRNA在相互作用网络中的功能和信号通路。测序数据中有51个circRNA差异表达:25个上调,26个下调。996个差异表达的lncRNA中,317个上调,679个下调;1161个差异表达的mRNA中,698个上调,463个下调。鉴定出33个差异表达的miRNA,上调的miRNA 18和下调的miRNA 15。筛选后,最终在lncRNA-miRNA-mRNA网络中构建了13个共表达的mRNA、15个lncRNA和3个miRNA,在circRNA-miRNA-mRNA网络中构建了159个mRNA、26个miRNA和34个circRNA。在lncRNA网络中,GO富集分析获得182个生物学过程、12个细胞成分和38个分子功能,主要与环核苷酸依赖性蛋白激酶活性和镁离子依赖性蛋白丝氨酸/苏氨酸磷酸酶活性等蛋白质和酶活性的调节有关。KEGG分析主要涉及叉头框O(FoxO)信号通路、apelin信号通路等。在circRNA富集结果中,获得353个生物学过程、52个细胞成分和45个分子功能,主要涉及钙通道调节、中性粒细胞活化、mRNA转运和代谢。KEGG主要涉及Wnt信号通路、apelin信号通路、Hippo信号通路、催产素信号通路等。本文为lncRNA和circRNA通过ceRNA机制介导NAION的发生发展提供了新思路。本研究为深入研究NAION的发病机制奠定了基础。

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