1 Reproduction Unit, Centro Clinico-Veterinario e Zootecnico-Sperimentale di Ateneo, Università degli Studi di Milano, Lodi, Italy.
2 Parco Tecnologico Padano, Lodi, Italy.
Cell Transplant. 2018 Jan;27(1):45-54. doi: 10.1177/0963689717724796.
Cell-derived microvesicles (MVs) are a recently discovered mechanism of cell-to-cell communication. Our previous data show that MVs secreted by equine amniotic mesenchymal-derived cells (AMCs) are involved in downregulation of proinflammatory genes in lipopolysaccharide-stressed equine tendon and endometrial cells. The aim of the present study was to evaluate whether AMC-MVs contain selected microRNAs (miRNAs) involved in inflammation. Two pools of cells, derived from 3 amniotic membranes each, and their respective MVs were collected. Small RNAs were extracted and deep sequenced, followed by miRNA in silico detection. The analysis identified 1,285 miRNAs, which were quantified both in AMCs and MVs. Among these miRNAs, 401 were classified as Equus caballus miRNAs, 257 were predicted by homology with other species (cow, sheep, and goat), and 627 were novel candidate miRNAs. Moreover, 146 miRNAs differentially expressed (DE) in AMCs and MVs were identified, 36 of which were known and the remaining were novel. Among the known DE miRNAs, 17 showed higher expression in MVs. Three of these were validated by real time polymerase chain reaction: eca-miR-26, eca-miR-146a, and eca-miR-223. Gene ontology analysis of validated targets showed that the DE miRNAs in cells and MVs could be involved both in immune system regulation by modulating interleukin signaling and in the inflammatory process. In conclusion, this study suggests a significant role of AMCs in modulating immune response through cell-cell communication via MV-shuttling miRNAs.
细胞衍生的微泡（MVs）是最近发现的细胞间通讯机制。我们之前的数据表明，马羊膜间充质衍生细胞（AMCs）分泌的 MV 参与下调脂多糖应激的马腱和子宫内膜细胞中的促炎基因。本研究旨在评估 AMCs-MVs 是否含有参与炎症的选定 microRNAs（miRNAs）。从 3 个羊膜中分别分离出 2 个细胞池及其各自的 MV。提取小 RNA 并进行深度测序，然后进行 miRNA 计算机检测。分析鉴定出 1285 个 miRNA，在 AMCs 和 MV 中均进行了定量。在这些 miRNA 中，401 个被归类为马 caballus miRNAs，257 个通过与其他物种（牛、绵羊和山羊）的同源性预测，627 个是新的候选 miRNA。此外，鉴定出在 AMCs 和 MV 中差异表达（DE）的 146 个 miRNA，其中 36 个是已知的，其余是新的。在已知的 DE miRNAs 中，有 17 个在 MV 中表达较高。其中 3 个通过实时聚合酶链反应进行验证：eca-miR-26、eca-miR-146a 和 eca-miR-223。经验证的靶基因的 GO 分析表明，细胞和 MV 中的 DE miRNAs 可能通过调节白细胞介素信号参与免疫系统调节和炎症过程。总之，这项研究表明 AMCs 通过 MV 转运的 miRNAs 进行细胞间通讯，在调节免疫反应方面发挥重要作用。
