间充质基质细胞衍生的细胞外囊泡通过 miR-181a-5p-PTEN-pSTAT5-SOCS1 轴重编程急性呼吸窘迫综合征模型中的巨噬细胞。
Mesenchymal stromal cells-derived extracellular vesicles reprogramme macrophages in ARDS models through the miR-181a-5p-PTEN-pSTAT5-SOCS1 axis.
机构信息
Wellcome-Wolfson Institute for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, Belfast, UK.
Department of Medicine, University of Vermont, Burlington, Vermont, USA.
出版信息
Thorax. 2023 Jun;78(6):617-630. doi: 10.1136/thoraxjnl-2021-218194. Epub 2022 Aug 10.
RATIONALE
A better understanding of the mechanism of action of mesenchymal stromal cells (MSCs) and their extracellular vesicles (EVs) is needed to support their use as novel therapies for acute respiratory distress syndrome (ARDS). Macrophages are important mediators of ARDS inflammatory response. Suppressor of cytokine signalling (SOCS) proteins are key regulators of the macrophage phenotype switch. We therefore investigated whether SOCS proteins are involved in mediation of the MSC effect on human macrophage reprogramming.
METHODS
Human monocyte-derived macrophages (MDMs) were stimulated with lipopolysaccharide (LPS) or plasma samples from patients with ARDS (these samples were previously classified into hypo-inflammatory and hyper-inflammatory phenotype) and treated with MSC conditioned medium (CM) or EVs. Protein expression was measured by Western blot. EV micro RNA (miRNA) content was determined by miRNA sequencing. In vivo: LPS-injured C57BL/6 mice were given EVs isolated from MSCs in which miR-181a had been silenced by miRNA inhibitor or overexpressed using miRNA mimic.
RESULTS
EVs were the key component of MSC CM responsible for anti-inflammatory modulation of human macrophages. EVs significantly reduced secretion of tumour necrosis factor-α and interleukin-8 by LPS-stimulated or ARDS plasma-stimulated MDMs and this was dependent on SOCS1. Transfer of miR-181a in EVs downregulated phosphatase and tensin homolog (PTEN) and subsequently activated phosphorylated signal transducer and activator of transcription 5 (pSTAT5) leading to upregulation of SOCS1 in macrophages. In vivo, EVs alleviated lung injury and upregulated pSTAT5 and SOCS1 expression in alveolar macrophages in a miR181-dependent manner. Overexpression of miR-181a in MSCs significantly enhanced therapeutic efficacy of EVs in this model.
CONCLUSION
miR-181a-PTEN-pSTAT5-SOCS1 axis is a novel pathway responsible for immunomodulatory effect of MSC EVs in ARDS.
背景
为了支持间充质基质细胞（MSCs）及其细胞外囊泡（EVs）作为急性呼吸窘迫综合征（ARDS）的新型治疗方法，需要更好地了解它们的作用机制。巨噬细胞是 ARDS 炎症反应的重要介质。细胞因子信号转导抑制因子（SOCS）蛋白是巨噬细胞表型转换的关键调节因子。因此，我们研究了 SOCS 蛋白是否参与调节 MSC 对人巨噬细胞重编程的作用。
方法
用脂多糖（LPS）或 ARDS 患者的血浆样本（这些样本先前被分类为低炎症和高炎症表型）刺激人单核细胞衍生的巨噬细胞（MDMs），并用 MSC 条件培养基（CM）或 EVs 处理。通过 Western blot 测量蛋白表达。通过 miRNA 测序确定 EV 微小 RNA（miRNA）含量。在体内：用沉默 miRNA 抑制剂或过表达 miRNA 模拟物的 MSC 中分离的 EV 处理 LPS 损伤的 C57BL/6 小鼠。
结果
EVs 是 MSC CM 中负责抗炎调节人巨噬细胞的关键成分。EVs 显著降低了 LPS 刺激或 ARDS 血浆刺激的 MDMs 的肿瘤坏死因子-α和白细胞介素-8 的分泌，这依赖于 SOCS1。EVs 中的 miR-181a 转移下调磷酸酶和张力蛋白同源物（PTEN），随后激活磷酸化信号转导和转录激活因子 5（pSTAT5），导致巨噬细胞中 SOCS1 的上调。在体内，EVs 以 miR181 依赖的方式减轻了肺损伤并上调了肺泡巨噬细胞中的 pSTAT5 和 SOCS1 表达。MSC 中 miR-181a 的过表达显著增强了该模型中 EVs 的治疗效果。
结论
miR-181a-PTEN-pSTAT5-SOCS1 轴是 MSC EVs 在 ARDS 中发挥免疫调节作用的新途径。
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