Distinct microRNA expression in human airway cells of asthmatic donors identifies a novel asthma-associated gene.
Airway inflammation is a hallmark of asthma, and suggests a dysregulation of homeostatic mechanisms. MicroRNAs (miRNAs) are key regulators of gene expression necessary for the proper function of cellular processes. We tested the hypothesis that differences between healthy and asthmatic subjects may be a result of distinct miRNA cellular profiles that lead to differential regulation of inflammatory genes. We collected human bronchial epithelial cells from seven healthy donors and seven patients with asthma, and profiled miRNA expression, using the Affymetrix (Santa Clara, CA) miRNA array platform. Results were confirmed according to quantitative RT-PCR on RNA isolated from 16 healthy and 16 asthmatic donors. We identified 66 miRNAs that were significantly different (≥ 1.5-fold; P ≤ 0.05) between the two groups, and validated three of them in epithelial cells from 16 asthmatic and 16 healthy subjects. Molecular network analysis indicated that putative targets were principally involved in regulating the expression of inflammatory pathway genes (P ≤ 10(-4)). Our analysis confirmed the prediction that the expression of IL-8, Cox2, and TNF-α was up-regulated in asthmatic cells, whereas the expression of IL-6 was lower compared with that in healthy control subjects. Network analysis was also used to identify a novel asthma-associated gene. The top-ranked predicted target of the highly down-regulated miRNA-203 in asthmatic cells was the aquaporin gene AQP4. Its expression was confirmed to be significantly higher in cells from patients with asthma. Overall, these data suggest that the heightened inflammatory pathway activation observed in patients with asthma may be attributed to underlying aberrant miRNA expression.
National Health and Environmental Effects Research Laboratory, Environmental Public Health Division, United States Environmental Protection Agency, Chapel Hill, NC 27514, USA. melaniejardim@gmail.com
Melanie J
MJ
Jardim
Lisa
L
Dailey
Robert
R
Silbajoris
David
D
Diaz-Sanchez
D000328:Adult; D051401:Aquaporin 4 / Q000235:genetics* / Q000378:metabolism; D001249:Asthma / Q000235:genetics* / Q000378:metabolism; D055745:Bronchioles / Q000473:pathology; D016022:Case-Control Studies; D002478:Cells, Cultured; D016000:Cluster Analysis; D005260:Female; D053263:Gene Regulatory Networks; D056726:Genetic Association Studies; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D008297:Male; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D020545:Respiratory Mucosa / Q000378:metabolism*; D014019:Tissue Donors; D059467:Transcriptome; D015854:Up-Regulation; D055815:Young Adult
C496030:AQP4 protein, human; D051401:Aquaporin 4; D018836:Inflammation Mediators; C546703:MIRN203 microRNA, human; D035683:MicroRNAs