嗅鞘细胞来源细胞外囊泡中 microRNAs 的差异表达及其靶基因分析
Differential Expression of microRNAs and Target Genes Analysis in Olfactory Ensheathing Cell-derived Extracellular Vesicles Olfactory Ensheathing Cells.
机构信息
Department of Orthopedics, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi Province, 710004, China.
Department of Orthopedics, Shaanxi Provincial People's Hospital, 256 Youyi West Road, Xi'an, 710068, Shaanxi, China.
出版信息
Curr Stem Cell Res Ther. 2024;19(1):116-125. doi: 10.2174/1574888X18666230418084900.
INTRODUCTION
Olfactory ensheathing cells (OECs) are important transplantable cells for the treatment of spinal cord injury. However, information on the mechanism of OEC-derived extracellular vesicles (EVs) in nerve repair is scarce.
METHODS
We cultured OECs and extracted the OEC-derived EVs, which were identified using a transmission electron microscope, nanoparticle flow cytometry, and western blotting. High throughput RNA sequencing of OECs and OEC-EVs was performed, and the differentially expressed microRNAs (miRNAs) (DERs) were analyzed by bioinformatics. The target genes of DERs were identified using miRWalk, miRDB, miRTarBase, and TargetScan databases. Gene ontology and KEGG mapper tools were used to analyze the predicted target genes. Subsequently, the STRING database and Cytoscape software platform were used to analyze and construct miRNA target genes' protein-protein interaction (PPI) network.
RESULTS
Overall, 206 miRNAs (105 upregulated and 101 downregulated) were differentially expressed in OEC-EVs (p < 0.05;|log2 (fold change)|>2). Six DERs (rno-miR-7a-5p, rno-miR-143-3p, rno-miR-182, rno-miR-214-3p, rno-miR-434-5p, rno-miR-543-3p) were significantly up-regulated , and a total of 974 miRNAs target genes were obtained. The target genes were mainly involved in biological processes such as regulation of cell size, positive regulation of cellular catabolic process and small GTPase-mediated signal transduction; positive regulation of genes involved in cellular components such as growth cone, site of polarized growth, and distal axon; and molecular functions such as small GTPase binding and Ras GTPase binding. In pathway analysis, target genes regulated by six DERs were mainly enriched in axon guidance, endocytosis, and Ras and cGMP-dependent protein kinase G signaling pathways. Finally, 19 hub genes were identified via the PPI network.
CONCLUSION
Our study provides a theoretical basis for treating nerve repair by OEC-derived EVs.
简介
嗅鞘细胞（OECs）是治疗脊髓损伤的重要可移植细胞。然而，关于 OEC 衍生的细胞外囊泡（EVs）在神经修复中的作用机制的信息还很缺乏。
方法
我们培养了 OECs 并提取了 OEC 衍生的 EVs，通过透射电子显微镜、纳米粒子流式细胞术和 Western blot 进行了鉴定。对 OECs 和 OEC-EVs 进行了高通量 RNA 测序，并通过生物信息学分析了差异表达的 microRNAs（miRNAs）（DERs）。使用 miRWalk、miRDB、miRTarBase 和 TargetScan 数据库鉴定 DER 的靶基因。GO 和 KEGG mapper 工具用于分析预测的靶基因。随后，使用 STRING 数据库和 Cytoscape 软件平台分析和构建 miRNA 靶基因的蛋白质-蛋白质相互作用（PPI）网络。
结果
总的来说，OEC-EVs 中有 206 个 miRNAs（105 个上调和 101 个下调）差异表达（p<0.05；|log2（fold change）|>2）。六个 DER（rno-miR-7a-5p、rno-miR-143-3p、rno-miR-182、rno-miR-214-3p、rno-miR-434-5p、rno-miR-543-3p）显著上调，共获得 974 个 miRNA 靶基因。靶基因主要参与细胞大小调节、细胞分解代谢过程的正调节和小 GTPase 介导的信号转导等生物学过程；参与生长锥、极化生长部位和远轴突等细胞成分的正调节；以及小 GTPase 结合和 Ras GTPase 结合等分子功能。在通路分析中，六个 DER 调节的靶基因主要富集在轴突导向、内吞作用和 Ras 和 cGMP 依赖性蛋白激酶 G 信号通路中。最后，通过 PPI 网络鉴定了 19 个枢纽基因。
结论
本研究为 OEC 衍生的 EV 治疗神经修复提供了理论依据。
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