Epithelial senescence in idiopathic pulmonary fibrosis is propagated by small extracellular vesicles

Respiratory Research

Research

Sabha Asghar¹  Lynne A. Murray¹  Catherine L. Overed-Sayer¹  Ken Grime²  David J. F. Smith³  Richard J. Hewitt³  Philip L. Molyneaux³  Susan Monkley⁴ 

[1] Bioscience COPD/IPF, Research & Early Development, Respiratory & Immunology, BioPharmaceuticals R&D, AstraZeneca, Cambridge, UK;Bioscience COPD/IPF, Research & Early Development, Respiratory & Immunology, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden;National Heart & Lung Institute, Imperial College London, London, UK;Royal Brompton & Harefield Hospitals, Guy’s & St Thomas’ NHS Foundation Trust, London, UK;Translational Sciences & Experimental Medicine, Research & Early Development, Respiratory & Immunology, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden;

关键词: Small extracellular vesicles;    Exosomes;    miRNA;    Senescence;    Idiopathic pulmonary fibrosis;   

DOI  :  10.1186/s12931-023-02333-5

 received in 2022-09-15, accepted in 2023-01-18,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundIdiopathic pulmonary fibrosis (IPF) is a chronic lung disease that affects 3 million people worldwide. Senescence and small extracellular vesicles (sEVs) have been implicated in the pathogenesis of IPF, although how sEVs promote disease remains unclear. Here, we profile sEVs from bronchial epithelial cells and determine small RNA (smRNA) content.MethodsConditioned media was collected and sEVs were isolated from normal human bronchial epithelial cells (NHBEs) and IPF-diseased human bronchial epithelial cells (DHBEs).ResultsIncreased sEV release from DHBEs compared to NHBEs (n = 4; p < 0.05) was detected by nanoparticle tracking analysis. NHBEs co-cultured with DHBE-derived sEVs for 72 h expressed higher levels of SA-β-Gal and γH2AX protein, p16 and p21 RNA and increased secretion of IL6 and IL8 proteins (all n = 6–8; p < 0.05). sEVs were also co-cultured with healthy air–liquid interface (ALI) cultures and similar results were observed, with increases in p21 and p16 gene expression and IL6 and IL8 (basal and apical) secretion (n = 6; p < 0.05). Transepithelial electrical resistance (TEER) measurements, a reflection of epithelial barrier integrity, were decreased upon the addition of DHBE-derived sEVs (n = 6; p < 0.05). smRNA-sequencing identified nineteen significantly differentially expressed miRNA in DHBE-derived sEVs compared to NHBE-derived sEVs, with candidate miRNAs validated by qPCR (all n = 5; p < 0.05). Four of these miRNAs were upregulated in NHBEs co-cultured with DHBE-derived sEVs and three in healthy ALI cultures co-cultured with DHBE-derived sEVs (n = 3–4; p < 0.05).ConclusionsThis data demonstrates that DHBE-derived sEVs transfer senescence to neighbouring healthy cells, promoting the disease state in IPF.

【 授权许可】

CC BY   
© The Author(s) 2023

【 图 表 】

Fig. 3

Fig. 1

Fig. 4

Fig. 2

Fig. 3

Fig. 2

Fig. 10