Asthma and lower airway disease
Transcriptional profiling identifies the long noncoding RNA plasmacytoma variant translocation (PVT1) as a novel regulator of the asthmatic phenotype in human airway smooth muscle

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Background

The mechanism underlying nonsevere and severe asthma remains unclear, although it is commonly associated with increased airway smooth muscle (ASM) mass. Long noncoding RNAs (lncRNAs) are known to be important in regulating healthy primary airway smooth muscle cells (ASMCs), whereas changed expression has been observed in CD8 T cells from patients with severe asthma.

Methods

Primary ASMCs were isolated from healthy subjects (n = 9) and patients classified as having nonsevere (n = 9) or severe (n = 9) asthma. ASMCs were exposed to dexamethasone and FCS. mRNA and lncRNA expression was measured by using a microarray and quantitative real-time PCR. Bioinformatic analysis was used to examine relevant biological pathways. Finally, the lncRNA plasmacytoma variant translocation 1 (PVT1) was inhibited by transfection of primary ASMCs with small interfering RNAs, and the effect on ASMC phenotype was examined.

Results

The mRNA expression profile was significantly different between patient groups after exposure to dexamethasone and FCS, and these were associated with biological pathways that might be relevant to the pathogenesis of asthma, including cellular proliferation and pathways associated with glucocorticoid activity. We also observed a significant change in lncRNA expression, yet the expression of only one lncRNA (PVT1) is decreased in patients with corticosteroid-sensitive nonsevere asthma and increased in patients with corticosteroid-insensitive severe asthma. Subsequent targeting studies demonstrated the importance of this lncRNA in controlling both proliferation and IL-6 release in ASMCs from patients with severe asthma.

Conclusions

lncRNAs are associated with the aberrant phenotype observed in ASMCs from asthmatic patients. Targeting PVT1 might be effective in reducing airway remodeling in asthmatic patients.

Key words

Asthma
airway smooth muscle
proliferation
IL-6
transcriptome
long noncoding RNA
PVT1

Abbreviations used

ASM
Airway smooth muscle
ASMC
Airway smooth muscle cell
BrdU
Bromodeoxyuridine
cRNA
Coding RNA
lncRNA
Long noncoding RNA
miRNA
MicroRNA
ncRNA
Noncoding RNA
PVT1
Plasmacytoma variant translocation
siRNA
Small interfering RNA

Cited by (0)

Supported by a fellowship from Imperial College London (to M.M.P.), grants from Asthma UK (08/041) and the Wellcome Trust (085935; to K.F.C.). This project was supported by the NIHR Respiratory Disease Biomedical Research Unit at the Royal Brompton and Harefield NHS Foundation Trust and Imperial College London. The views expressed in this publication are those of the authors(s) and not necessarily those of the National Health Service, National Institute for Health Research (NIHR), or Department of Health. K.F.C. is a Senior Investigator of the NIHR. M.M.P. was an Imperial College Research Fellow. M.M.P., I.M.A., and K.F.C. are members of the Interuniversity Attraction Poles Program-Belgian State-Belgian Science Policy- project P7/30.

Disclosure of potential conflict of interest: The authors declare that they have no relevant conflicts of interest.