Mesenchymal stem cells-derived exosomes carrying microRNA-30b confer protection against pulmonary fibrosis by downregulating Runx1 via Spred2

IF 2.3 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Genetics and Genomics Pub Date : 2024-03-13 DOI:10.1007/s00438-024-02116-7

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Abstract

Idiopathic pulmonary fibrosis (IPF) is a chronic pulmonary fibrosis disease that is fatal. Mesenchymal stem cells (MSCs)-secreted exosomes (exos) have been linked to improving PF. Moreover, exosomal microRNAs (miRs) can control the growth of numerous diseases, including lung disorders. Our bioinformatics analysis showed that miR-30b was downregulated in tissue samples from surgical remnants of biopsies or lungs explanted from patients with IPF who underwent pulmonary transplantation. This suggests that miR-30b plays an important role in both the pathogenesis and treatment of IPF. Herein, this research was designed to ascertain the mechanism of MSCs-exos-packaged miR-30b in alleviating PF. The serum was harvested from idiopathic PF (IPF) patients with interstitial pneumonia caused by dermatomyositis and the MLE12 lung epithelial cell fibrosis model was built with TGF-β1 (10 ng/mL), followed by miR-30b expression determination. TGF-β1-stimulated MLE12 cells were co-incubated with exos from MSCs with or without Spred2 or Runx1 overexpression, followed by measurement of cell viability and apoptosis. After establishing the IPF mouse model with bleomycin and injecting exos and/or silencing and overexpressing adenovirus vectors, fibrosis evaluation was conducted. In mice and cells, the expression of TGF-β1, TNF-α, and IL-1β was tested via ELISA, and the levels of E-cad, ZO-1, α-SMA, and collagen type I via western blot analysis. The promoters of miR-30b, Runx1, and Spred2 were investigated. miR-30b was downregulated in the serum of IPF patients and TGF-β1-stimulated MLE12 cells. Mechanistically, miR-30b inhibited Spred2 transcription by negatively targeting Runx1. MSCs-exos or MSCs-exo-miR-30b decreased the apoptosis, inflammation, and fibrosis while increasing their viability in TGF-β1-stimulated MLE12 cells, which was annulled by overexpressing Runx1 or Spred2. Exo-miR-30b decreased Runx1 expression to downregulate Spred2, reducing fibrosis and inflammation in IPF mice. Our results indicated that MSCs-exos-encapsulated miR-30b had a potential function to inhibit PF and part of its function may be achieved by targeting RUNX1 to reduce the Spred2 transcription level. Moreover, this work offered evidence and therapeutic targets for therapeutic strategies for managing clinical PF in patients.

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间充质干细胞衍生的携带 microRNA-30b 的外泌体通过 Spred2 下调 Runx1，从而防止肺纤维化

摘要特发性肺纤维化（IPF）是一种致命的慢性肺纤维化疾病。间充质干细胞（MSCs）分泌的外泌体（exos）与改善肺纤维化有关。此外，外泌体微RNA（miRs）可控制包括肺部疾病在内的多种疾病的生长。我们的生物信息学分析表明，在接受肺移植手术的 IPF 患者的手术残留活检组织样本或肺切除组织样本中，miR-30b 出现下调。这表明，miR-30b 在 IPF 的发病机制和治疗中发挥着重要作用。因此，本研究旨在确定间充质干细胞-外胚层包裹的 miR-30b 在缓解 PF 中的作用机制。研究人员采集了由皮肌炎引起间质性肺炎的特发性 PF（IPF）患者的血清，并用 TGF-β1（10 ng/mL）建立了 MLE12 肺上皮细胞纤维化模型，随后测定了 miR-30b 的表达。TGF-β1刺激的MLE12细胞与Spred2或Runx1过表达或未表达的间充质干细胞外泌体共同孵育，然后测定细胞活力和凋亡。在用博来霉素建立 IPF 小鼠模型并注射外泌体和/或沉默和过表达腺病毒载体后，进行了纤维化评估。在小鼠和细胞中，通过 ELISA 检测了 TGF-β1、TNF-α 和 IL-1β 的表达，通过 Western 印迹分析检测了 E-cad、ZO-1、α-SMA 和胶原 I 型的水平。在 IPF 患者血清和 TGF-β1 刺激的 MLE12 细胞中，miR-30b 被下调。从机制上讲，miR-30b通过负靶向Runx1抑制了Spred2的转录。间充质干细胞-exos或间充质干细胞-exo-miR-30b可减少TGF-β1刺激的MLE12细胞的凋亡、炎症和纤维化，同时提高其存活率。Exo-miR-30b 可减少 Runx1 的表达，从而下调 Spred2，减轻 IPF 小鼠的纤维化和炎症。我们的研究结果表明，间充质干细胞外包被的miR-30b具有抑制PF的潜在功能，其部分功能可能是通过靶向RUNX1以降低Spred2的转录水平来实现的。此外，这项研究还为临床PF患者的治疗策略提供了证据和治疗靶点。

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来源期刊

Molecular Genetics and Genomics 生物-生化与分子生物学

CiteScore

5.10

自引率

3.20%

发文量

134

审稿时长

1 months

期刊介绍： Molecular Genetics and Genomics (MGG) publishes peer-reviewed articles covering all areas of genetics and genomics. Any approach to the study of genes and genomes is considered, be it experimental, theoretical or synthetic. MGG publishes research on all organisms that is of broad interest to those working in the fields of genetics, genomics, biology, medicine and biotechnology. The journal investigates a broad range of topics, including these from recent issues: mechanisms for extending longevity in a variety of organisms; screening of yeast metal homeostasis genes involved in mitochondrial functions; molecular mapping of cultivar-specific avirulence genes in the rice blast fungus and more.