Comparative Analysis of microRNA Expression Profiles of Exosome-Mimetic Vesicles, Exosomes, and Originating Human Bone Marrow Mesenchymal Stem Cells.

Stem cells and development Pub Date : 2025-07-01 Epub Date: 2025-06-17 DOI:10.1089/scd.2025.0005
Congya Zhang, Jie Yu, Shuhong Chen, Guyan Wang
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Abstract

Exosomes derived from mesenchymal stem cells (MSCs) show therapeutic potential despite limited yield. In contrast, the yield of exosome-mimetic vesicles (EMVs), which share a similar structure and size with exosomes is high. Previous studies have compared their proteomic profiles, and the microRNA (miRNA) expression signatures remain unexplored. EMVs from human bone marrow MSCs were isolated through continuous extrusion and exosomes were isolated from the supernatant via differential ultracentrifugation. miRNA sequencing was performed using high-throughput sequencing, and the miRNA expression profiles of MSC-EMVs, MSC-exosomes, and MSCs were compared. Following the comparison of differentially expressed miRNAs in MSC-EMVs and MSC-exosomes, target gene prediction and functional enrichment analyses were performed. Furthermore, a trend analysis was conducted on the miRNA expression levels in the three groups to further explore the relationship between miRNA expression levels. Our study confirmed that EMVs could be stably produced and that their yield was approximately 100-fold higher than that of exosomes. A total of 763 known miRNAs were identified through comparison using the miRBase library. The miRNAs in EMVs and exosomes overlapped with those in MSCs; however, EMVs shared more miRNAs with the parent cells. Comparative analysis identified 21 upregulated and 17 downregulated miRNAs in EMVs versus exosomes, while trend analysis revealed 108 miRNAs preferentially expressed in MSCs and EMVs. Functional enrichment of differentially expressed miRNAs provides new insights for EMV-based therapies. Importantly, we demonstrated that both MSC-EMVs and MSC-exosomes significantly attenuated LPS-induced inflammation in THP-1 macrophages by modulating cytokine secretion (ELISA), suppressing iNOS expression (immunofluorescence), and inhibiting NF-κB activation (western blot). In an lipopolysaccharide (LPS)-induced acute kidney injury model, both vesicle types effectively reduced renal inflammation and tissue damage (histopathology and protein analysis). Our findings not only present the first comprehensive miRNA profiling comparison between MSC-derived EMVs and exosomes but also validate their comparable anti-inflammatory efficacy, supporting EMVs as a viable high-yield alternative for cell-free therapies.

模拟外泌体囊泡、外泌体和原源人骨髓间充质干细胞microRNA表达谱的比较分析。
来自间充质干细胞(MSCs)的外泌体尽管产量有限,但仍显示出治疗潜力。相比之下,与外泌体具有相似结构和大小的外泌体模拟囊泡(emv)的产量很高。先前的研究已经比较了它们的蛋白质组学特征,而microRNA (miRNA)的表达特征仍未被探索。通过连续挤压从人骨髓间充质干细胞中分离emv,并通过差动超离心从上清中分离外泌体。采用高通量测序进行miRNA测序,比较msc - emv、msc -外泌体和MSCs的miRNA表达谱。通过比较msc - emv和msc -外泌体中差异表达的mirna,进行靶基因预测和功能富集分析。进一步对三组患者的miRNA表达水平进行趋势分析,进一步探讨miRNA表达水平之间的关系。我们的研究证实,emv可以稳定地产生,其产量大约是外泌体的100倍。通过miRBase文库的比较,共鉴定出763个已知的mirna。emv和外泌体中的mirna与MSCs中的mirna重叠;然而,emv与亲本细胞共享更多的mirna。对比分析发现,emv与外泌体中有21个上调的mirna和17个下调的mirna,而趋势分析显示,有108个mirna优先在MSCs和emv中表达。差异表达mirna的功能富集为基于emv的治疗提供了新的见解。重要的是,我们证明了msc - emv和msc -外泌体通过调节细胞因子分泌(ELISA)、抑制iNOS表达(免疫荧光)和抑制NF-κB激活(western blot)显著减轻lps诱导的THP-1巨噬细胞炎症。在脂多糖(LPS)诱导的急性肾损伤模型中,两种囊泡类型都有效地减轻了肾脏炎症和组织损伤(组织病理学和蛋白质分析)。我们的研究结果不仅首次全面比较了msc衍生的emv和外泌体之间的miRNA谱,而且验证了它们的抗炎功效,支持emv作为无细胞治疗的可行高产替代方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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