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Mechanical stretch differentially regulates the expression of specific miRNA in the extracellular vesicles released from lung epithelial cells

ABSTRACT: Mechanical force is critical for lung development. In this study we identified specific EV-miRNAs released from the mouse epithelial cell line in response to mechanical stretch involve in lung development. In utero fetal lung experiences significant continuous transpulmonary pressure as a result of epithelial secretion in to the airway lumen, and periodic fetal breathing movement that move the fluid along the developing airway. Mechanical force is important factors for fetal lung development. However, the effect of mechanical force on the functions of lung cells is not known precisely. Extracellular vesicles –microRNAs (EV-miRNA) are increasingly recognized as a new mode of cell-to-cell communication. miRNA is well known as a regulator of physio-pathological process. In this study, we used oligonucleotide microarray technology to investigate miRNA expression in EV-released from mouse lung epithelial cell MLE12 after exposed to 10% cyclic or 5% continuous stretch. Analysis of microarray data identified 9 and 33 miRNAs significantly differentially expressed by the cyclic and continuous stretch respectively. Several differentially expressed miRNAs were reported dynamically expressed in mouse developing lung. miRNAs associated with important transcription factors for cell function and key signaling pathways for fetal lung development also identified in this study. We conclude that mechanical signals differentially regulate the expression of specific EV/miRNAs in MLE12 are important for intercellular communication during lung fetal development.

ORGANISM(S): Mus musculus synthetic construct

PROVIDER: GSE131645 | GEO | 2019/05/23

REPOSITORIES: GEO

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Project description:Acute respiratory distress syndrome (ARDS) is a catastrophic form of acute lung injury (ALI). The necessity for mechanical ventilation (MV) renders patients at risk for ventilator induced lung injury (VILI). Exposure to repetitive cyclic stretch (CS) and/or over-inflation exacerbates injury. Reducing tidal volume (VT) is the only therapeutic strategy shown to mitigate morbidity and mortality. Cyclic stretch has been shown to differentially regulate gene expression in part through the activation of mammalian mitogen-activated protein kinase (MAPK). Although these studies have shown both molecular and cellular alterations, no unifying hypothesis to explain MV-induced lung injury has emerged. In the current study, we hypothesized that coordinated expression of cyclic stretch (CS)-responsive genes relies on the presence of common CS-sensitive regulatory elements. To identify CS-responsive genes, we undertook a comparative examination of the gene expression profile of human bronchial epithelial airway (Beas-2B) cells in response to various injurious stimuli involved in the pathogenesis of acute lung injury (ALI)/Ventilator induced lung injury (VILI): cyclic stretch, tumor necrosis factor alpha (TNF-a), and lipopolysaccharide (LPS). Experiment Overall Design: Human Bronchial Epithelial Cells (Beas-B2) cells grown on silicon elastic plates coated with Type I collagen (Flexercell International, McKeesport, PA) were exposed to six regiments for 4 h: 1) control (static, [control]); 2) mechanical stretch (25 PKa, 30 cycles per min, [stretch]); 3) LPS (1 mcg/ml [LPS]); 4) TNF-Î± (20 ng/ml; [TNF]); 5) mechanical stretch plus LPS [LPS+S], and 6) mechanical stretch plus TNF-Î± [TNF+S]. Total RNA (duplicate experiments) was extracted using TRIZOL reagent (as per manufactures specifications) and purified using Qiagen mRNA purification Kit (as per manufacturers specifications). mRNA was hybridized to Affymetrix Human U133plus2.0 chips. Probe based analysis, background reduction, and quantile data normalization was performed in MeV 4.0 of TM4 using Robust Multi-array Average (RMA).

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Mechanical stretch differentially regulates the expression of specific miRNA in the extracellular vesicles released from lung epithelial cells

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