Unit of Radiation Medicine, Department of Radiobiology and Radiohygiene, National Public Health Centre, 1097 Budapest, Hungary.
Doctoral School of Pathological Sciences, Semmelweis University, 1085 Budapest, Hungary.
Int J Mol Sci. 2023 May 11;24(10):8607. doi: 10.3390/ijms24108607.
Extracellular vesicles (EVs), through their cargo, are important mediators of bystander responses in the irradiated bone marrow (BM). MiRNAs carried by EVs can potentially alter cellular pathways in EV-recipient cells by regulating their protein content. Using the CBA/Ca mouse model, we characterised the miRNA content of BM-derived EVs from mice irradiated with 0.1 Gy or 3 Gy using an nCounter analysis system. We also analysed proteomic changes in BM cells either directly irradiated or treated with EVs derived from the BM of irradiated mice. Our aim was to identify key cellular processes in the EV-acceptor cells regulated by miRNAs. The irradiation of BM cells with 0.1 Gy led to protein alterations involved in oxidative stress and immune and inflammatory processes. Oxidative stress-related pathways were also present in BM cells treated with EVs isolated from 0.1 Gy-irradiated mice, indicating the propagation of oxidative stress in a bystander manner. The irradiation of BM cells with 3 Gy led to protein pathway alterations involved in the DNA damage response, metabolism, cell death and immune and inflammatory processes. The majority of these pathways were also altered in BM cells treated with EVs from mice irradiated with 3 Gy. Certain pathways (cell cycle, acute and chronic myeloid leukaemia) regulated by miRNAs differentially expressed in EVs isolated from mice irradiated with 3 Gy overlapped with protein pathway alterations in BM cells treated with 3 Gy EVs. Six miRNAs were involved in these common pathways interacting with 11 proteins, suggesting the involvement of miRNAs in the EV-mediated bystander processes. In conclusion, we characterised proteomic changes in directly irradiated and EV-treated BM cells, identified processes transmitted in a bystander manner and suggested miRNA and protein candidates potentially involved in the regulation of these bystander processes.
细胞外囊泡 (EV) 通过其携带的货物，是辐照骨髓 (BM) 中旁观者反应的重要介质。EV 携带的 miRNAs 可以通过调节其蛋白质含量来潜在改变 EV 受体细胞中的细胞途径。我们使用 CBA/Ca 小鼠模型，使用 nCounter 分析系统，对 0.1Gy 或 3Gy 辐照小鼠的 BM 衍生 EV 中的 miRNA 含量进行了表征。我们还分析了直接辐照或用辐照小鼠 BM 衍生 EV 处理的 BM 细胞中的蛋白质变化。我们的目的是确定 EV 受体细胞中受 miRNAs 调节的关键细胞过程。0.1Gy 辐照 BM 细胞导致与氧化应激以及免疫和炎症过程相关的蛋白质改变。用来自 0.1Gy 辐照小鼠的 EV 处理的 BM 细胞中也存在与氧化应激相关的途径，表明氧化应激以旁观者方式传播。3Gy 辐照 BM 细胞导致与 DNA 损伤反应、代谢、细胞死亡以及免疫和炎症过程相关的蛋白质途径改变。用来自 3Gy 辐照小鼠的 EV 处理的 BM 细胞中也改变了这些途径中的大多数。某些由在 3Gy 辐照的 EV 中差异表达的 miRNAs 调节的途径（细胞周期、急性和慢性髓性白血病）与用 3Gy EV 处理的 BM 细胞中的蛋白质途径改变重叠。有 6 个 miRNAs 参与了这些与 EV 介导的旁观者过程有关的共同途径，与 11 个蛋白质相互作用，提示 miRNA 参与了 EV 介导的旁观者过程。总之，我们对直接辐照和 EV 处理的 BM 细胞中的蛋白质组学变化进行了表征，确定了以旁观者方式传递的过程，并提出了可能参与这些旁观者过程调节的 miRNA 和蛋白质候选物。
