Yin Xin, Yi Jialian, Mao Fugang, Tang Qisheng, Zhang Xinyu, Yang Xiaoyu, Xie Hongqing, Wang Linping, Sun Shuifen, Yu Xin, Liu Jie, Jiang Lihong
Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, China.
The Affiliated Hospital of Kunming University of Science and Technology, Kunming, China.
Front Genet. 2025 Jan 15;15:1407671. doi: 10.3389/fgene.2024.1407671. eCollection 2024.
This study aimed to investigate the impact of low-intensity pulsed ultrasound (LIPUS) treatment on the miRNA and mRNA profiles of stem cell-derived extracellular vesicles (EVs). Specifically, it sought to identify key miRNAs and their target mRNAs associated with enhanced therapeutic efficacy in LIPUS-treated stem cell-derived EVs.
Utilizing miRNA deep-sequencing data from the Gene Expression Omnibus database, differential gene analysis was performed. MiRNA-mRNA target analysis, functional and pathway enrichment analysis, protein-protein interaction network construction, and hub gene identification were conducted. Validation of differentially expressed miRNAs was performed via RT-qPCR in human umbilical cord mesenchymal stem cells (hUC-MSCs) treated with LIPUS.
Ten differentially expressed miRNAs were identified, with six upregulated and four downregulated miRNAs in LIPUS-treated stem cell-derived EVs. Functional enrichment analysis revealed involvement in biological processes such as regulation of metabolic processes, cellular component organization, and response to stress, as well as signaling pathways like cell cycle, MAPK signaling, and Hippo signaling. Protein-protein interaction network analysis identified key hub genes including MYC, GAPDH, HSP90AA1, EP300, JUN, PTEN, DAC1, STAT3, HSPA8, and HIF1A associated with LIPUS treatment. RT-qPCR validation confirmed differential expression of selected miRNAs (hsa-miR-933, hsa-miR-3943, hsa-miR-4633-5p, hsa-miR-592, hsa-miR-659-5p, hsa-miR-4766-3p) in LIPUS-treated hUC-MSCs.
This study sheds light on the potential therapeutic mechanisms underlying LIPUS-treated stem cell-derived EVs. The identified differentially expressed miRNAs and their potential target mRNAs offer valuable insights into the biological processes influenced by LIPUS treatment. While further investigation is necessary to validate their roles as therapeutic targets, this study lays the groundwork for future research on optimizing SC-EV therapy with LIPUS preconditioning.
本研究旨在探讨低强度脉冲超声（LIPUS）治疗对干细胞衍生细胞外囊泡（EVs）的miRNA和mRNA谱的影响。具体而言，它试图确定与LIPUS处理的干细胞衍生EVs中增强的治疗效果相关的关键miRNA及其靶mRNA。
利用基因表达综合数据库中的miRNA深度测序数据进行差异基因分析。进行了miRNA-mRNA靶标分析、功能和通路富集分析、蛋白质-蛋白质相互作用网络构建以及枢纽基因鉴定。通过RT-qPCR在接受LIPUS处理的人脐带间充质干细胞（hUC-MSCs）中对差异表达的miRNA进行验证。
鉴定出10个差异表达的miRNA，在LIPUS处理的干细胞衍生EVs中有6个miRNA上调，4个miRNA下调。功能富集分析显示其参与了代谢过程调节、细胞成分组织和应激反应等生物过程，以及细胞周期、MAPK信号传导和Hippo信号传导等信号通路。蛋白质-蛋白质相互作用网络分析确定了与LIPUS治疗相关的关键枢纽基因，包括MYC、GAPDH、HSP90AA1、EP300、JUN、PTEN、DAC1、STAT3、HSPA8和HIF1A。RT-qPCR验证证实了在接受LIPUS处理的hUC-MSCs中所选miRNA（hsa-miR-933、hsa-miR-3943、hsa-miR-4633-5p、hsa-miR-592、hsa-miR-659-5p、hsa-miR-4766-3p）的差异表达。
本研究揭示了LIPUS处理的干细胞衍生EVs潜在的治疗机制。鉴定出的差异表达miRNA及其潜在的靶mRNA为LIPUS治疗影响的生物过程提供了有价值的见解。虽然需要进一步研究来验证它们作为治疗靶点的作用，但本研究为未来利用LIPUS预处理优化SC-EV治疗的研究奠定了基础。
