Limits...
Persistence of smoking-induced dysregulation of miRNA expression in the small airway epithelium despite smoking cessation.

Wang G, Wang R, Strulovici-Barel Y, Salit J, Staudt MR, Ahmed J, Tilley AE, Yee-Levin J, Hollmann C, Harvey BG, Kaner RJ, Mezey JG, Sridhar S, Pillai SG, Hilton H, Wolff G, Bitter H, Visvanathan S, Fine JS, Stevenson CS, Crystal RG - PLoS ONE (2015)

Bottom Line: There were significant differences in the expression of 34 miRNAs between healthy smokers and healthy nonsmokers (p<0.01, fold-change >1.5), with functions associated with lung development, airway epithelium differentiation, inflammation and cancer.After quitting smoking for 3 months, 12 out of the 34 miRNAs did not return to normal levels, with Wnt/β-catenin signaling pathway being the top identified enriched pathway of the target genes of the persistent dysregulated miRNAs.In the context that many of these persistent smoking-dependent miRNAs are associated with differentiation, inflammatory diseases or lung cancer, it is likely that persistent smoking-related changes in SAE miRNAs play a role in the subsequent development of these disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Medicine, Weill Cornell Medical College, New York, New York, United States of America.

ABSTRACT
Even after quitting smoking, the risk of the development of chronic obstructive pulmonary disease (COPD) and lung cancer remains significantly higher compared to healthy nonsmokers. Based on the knowledge that COPD and most lung cancers start in the small airway epithelium (SAE), we hypothesized that smoking modulates miRNA expression in the SAE linked to the pathogenesis of smoking-induced airway disease, and that some of these changes persist after smoking cessation. SAE was collected from 10th to 12th order bronchi using fiberoptic bronchoscopy. Affymetrix miRNA 2.0 arrays were used to assess miRNA expression in the SAE from 9 healthy nonsmokers and 10 healthy smokers, before and after they quit smoking for 3 months. Smoking status was determined by urine nicotine and cotinine measurement. There were significant differences in the expression of 34 miRNAs between healthy smokers and healthy nonsmokers (p<0.01, fold-change >1.5), with functions associated with lung development, airway epithelium differentiation, inflammation and cancer. After quitting smoking for 3 months, 12 out of the 34 miRNAs did not return to normal levels, with Wnt/β-catenin signaling pathway being the top identified enriched pathway of the target genes of the persistent dysregulated miRNAs. In the context that many of these persistent smoking-dependent miRNAs are associated with differentiation, inflammatory diseases or lung cancer, it is likely that persistent smoking-related changes in SAE miRNAs play a role in the subsequent development of these disorders.

Show MeSH

Related in: MedlinePlus

Target genes of smoking cessation persistently altered miRNAs in the human SAE.A: Numbers of predicted target genes for each of the persistent miRNAs. The target genes prediction was based on Target Scan 6.2 database. Only genes that are expressed in human SAE were used for the analysis. Blue, target genes of persistent-down-regulated miRNAs. Red, target genes of persistent-up-regulated miRNA. The persistently dysregulated miRNAs with the highest target gene number were miR-218 and miR-133a and miR-133b targets. B: Top 10 enriched canonical pathways in the target genes of the smoking cessation persistent miRNAs. The analysis was performed using Ingenuity Pathway Analysis software. X axis,-log of p value, Fisher's exact test. The ratio of genes that were targeted by the smoking cessation persistent-miRNA in each pathway are indicated. C: Wnt/β-catenin signaling pathway associated with persistently dysregulated miRNAs despite smoking cessation. The Wnt pathway genes that are targets of the smoking cessation persistent miRNAs are highlighted by yellow. The correspondent miRNAs are marked red, with the number of red dots corresponding to the number of miRNAs targeted toward each gene. Many Wnt pathway ligands, receptors, effectors and regulators are potential targets of the smoking cessation persistent miRNAs. Abbreviations: IGF-1, insulin-like growth factor 1; ERK5, Extracellular signal-regulated kinase 5; MAPK, mitogen-activated protein kinase.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4401720&req=5

pone.0120824.g004: Target genes of smoking cessation persistently altered miRNAs in the human SAE.A: Numbers of predicted target genes for each of the persistent miRNAs. The target genes prediction was based on Target Scan 6.2 database. Only genes that are expressed in human SAE were used for the analysis. Blue, target genes of persistent-down-regulated miRNAs. Red, target genes of persistent-up-regulated miRNA. The persistently dysregulated miRNAs with the highest target gene number were miR-218 and miR-133a and miR-133b targets. B: Top 10 enriched canonical pathways in the target genes of the smoking cessation persistent miRNAs. The analysis was performed using Ingenuity Pathway Analysis software. X axis,-log of p value, Fisher's exact test. The ratio of genes that were targeted by the smoking cessation persistent-miRNA in each pathway are indicated. C: Wnt/β-catenin signaling pathway associated with persistently dysregulated miRNAs despite smoking cessation. The Wnt pathway genes that are targets of the smoking cessation persistent miRNAs are highlighted by yellow. The correspondent miRNAs are marked red, with the number of red dots corresponding to the number of miRNAs targeted toward each gene. Many Wnt pathway ligands, receptors, effectors and regulators are potential targets of the smoking cessation persistent miRNAs. Abbreviations: IGF-1, insulin-like growth factor 1; ERK5, Extracellular signal-regulated kinase 5; MAPK, mitogen-activated protein kinase.

Mentions: To further assess the roles of smoking cessation persistent miRNAs in the SAE, the functions of their target genes were explored. Only those target genes known to be expressed in SAE were used for analysis [21]. There were 2,112 genes (excluding duplicates) that were predicted-targets of the smoking cessation persistent-miRNAs. Among these genes, 1763 were expressed in the human SAE. Interestingly, many of the target genes were targets of miR-218 and miR-133a/b (Fig 4A). By pathway analysis, Wnt/β-catenin signaling, cardiac β-adrenergic signaling and protein kinase A signaling were found to be the top 3 canonical pathways (ranked by p value) enriched in the target genes of smoking cessation persistent-miRNAs (Fig 4B and S3 Table). The Wnt/ β-catenin signaling pathway was the top enriched pathway, consistent with smoking being associated with dysregulated differentiation, lung cancer and inflammation. Remarkably, 24% of Wnt pathway genes were predicted to be targets of smoking cessation miRNAs, with some Wnt pathway genes targeted by multiple miRNAs, including Wnt ligands, receptors, regulators and effectors (Fig 4C).


Persistence of smoking-induced dysregulation of miRNA expression in the small airway epithelium despite smoking cessation.

Wang G, Wang R, Strulovici-Barel Y, Salit J, Staudt MR, Ahmed J, Tilley AE, Yee-Levin J, Hollmann C, Harvey BG, Kaner RJ, Mezey JG, Sridhar S, Pillai SG, Hilton H, Wolff G, Bitter H, Visvanathan S, Fine JS, Stevenson CS, Crystal RG - PLoS ONE (2015)

Target genes of smoking cessation persistently altered miRNAs in the human SAE.A: Numbers of predicted target genes for each of the persistent miRNAs. The target genes prediction was based on Target Scan 6.2 database. Only genes that are expressed in human SAE were used for the analysis. Blue, target genes of persistent-down-regulated miRNAs. Red, target genes of persistent-up-regulated miRNA. The persistently dysregulated miRNAs with the highest target gene number were miR-218 and miR-133a and miR-133b targets. B: Top 10 enriched canonical pathways in the target genes of the smoking cessation persistent miRNAs. The analysis was performed using Ingenuity Pathway Analysis software. X axis,-log of p value, Fisher's exact test. The ratio of genes that were targeted by the smoking cessation persistent-miRNA in each pathway are indicated. C: Wnt/β-catenin signaling pathway associated with persistently dysregulated miRNAs despite smoking cessation. The Wnt pathway genes that are targets of the smoking cessation persistent miRNAs are highlighted by yellow. The correspondent miRNAs are marked red, with the number of red dots corresponding to the number of miRNAs targeted toward each gene. Many Wnt pathway ligands, receptors, effectors and regulators are potential targets of the smoking cessation persistent miRNAs. Abbreviations: IGF-1, insulin-like growth factor 1; ERK5, Extracellular signal-regulated kinase 5; MAPK, mitogen-activated protein kinase.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401720&req=5

pone.0120824.g004: Target genes of smoking cessation persistently altered miRNAs in the human SAE.A: Numbers of predicted target genes for each of the persistent miRNAs. The target genes prediction was based on Target Scan 6.2 database. Only genes that are expressed in human SAE were used for the analysis. Blue, target genes of persistent-down-regulated miRNAs. Red, target genes of persistent-up-regulated miRNA. The persistently dysregulated miRNAs with the highest target gene number were miR-218 and miR-133a and miR-133b targets. B: Top 10 enriched canonical pathways in the target genes of the smoking cessation persistent miRNAs. The analysis was performed using Ingenuity Pathway Analysis software. X axis,-log of p value, Fisher's exact test. The ratio of genes that were targeted by the smoking cessation persistent-miRNA in each pathway are indicated. C: Wnt/β-catenin signaling pathway associated with persistently dysregulated miRNAs despite smoking cessation. The Wnt pathway genes that are targets of the smoking cessation persistent miRNAs are highlighted by yellow. The correspondent miRNAs are marked red, with the number of red dots corresponding to the number of miRNAs targeted toward each gene. Many Wnt pathway ligands, receptors, effectors and regulators are potential targets of the smoking cessation persistent miRNAs. Abbreviations: IGF-1, insulin-like growth factor 1; ERK5, Extracellular signal-regulated kinase 5; MAPK, mitogen-activated protein kinase.
Mentions: To further assess the roles of smoking cessation persistent miRNAs in the SAE, the functions of their target genes were explored. Only those target genes known to be expressed in SAE were used for analysis [21]. There were 2,112 genes (excluding duplicates) that were predicted-targets of the smoking cessation persistent-miRNAs. Among these genes, 1763 were expressed in the human SAE. Interestingly, many of the target genes were targets of miR-218 and miR-133a/b (Fig 4A). By pathway analysis, Wnt/β-catenin signaling, cardiac β-adrenergic signaling and protein kinase A signaling were found to be the top 3 canonical pathways (ranked by p value) enriched in the target genes of smoking cessation persistent-miRNAs (Fig 4B and S3 Table). The Wnt/ β-catenin signaling pathway was the top enriched pathway, consistent with smoking being associated with dysregulated differentiation, lung cancer and inflammation. Remarkably, 24% of Wnt pathway genes were predicted to be targets of smoking cessation miRNAs, with some Wnt pathway genes targeted by multiple miRNAs, including Wnt ligands, receptors, regulators and effectors (Fig 4C).

Bottom Line: There were significant differences in the expression of 34 miRNAs between healthy smokers and healthy nonsmokers (p<0.01, fold-change >1.5), with functions associated with lung development, airway epithelium differentiation, inflammation and cancer.After quitting smoking for 3 months, 12 out of the 34 miRNAs did not return to normal levels, with Wnt/β-catenin signaling pathway being the top identified enriched pathway of the target genes of the persistent dysregulated miRNAs.In the context that many of these persistent smoking-dependent miRNAs are associated with differentiation, inflammatory diseases or lung cancer, it is likely that persistent smoking-related changes in SAE miRNAs play a role in the subsequent development of these disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetic Medicine, Weill Cornell Medical College, New York, New York, United States of America.

ABSTRACT
Even after quitting smoking, the risk of the development of chronic obstructive pulmonary disease (COPD) and lung cancer remains significantly higher compared to healthy nonsmokers. Based on the knowledge that COPD and most lung cancers start in the small airway epithelium (SAE), we hypothesized that smoking modulates miRNA expression in the SAE linked to the pathogenesis of smoking-induced airway disease, and that some of these changes persist after smoking cessation. SAE was collected from 10th to 12th order bronchi using fiberoptic bronchoscopy. Affymetrix miRNA 2.0 arrays were used to assess miRNA expression in the SAE from 9 healthy nonsmokers and 10 healthy smokers, before and after they quit smoking for 3 months. Smoking status was determined by urine nicotine and cotinine measurement. There were significant differences in the expression of 34 miRNAs between healthy smokers and healthy nonsmokers (p<0.01, fold-change >1.5), with functions associated with lung development, airway epithelium differentiation, inflammation and cancer. After quitting smoking for 3 months, 12 out of the 34 miRNAs did not return to normal levels, with Wnt/β-catenin signaling pathway being the top identified enriched pathway of the target genes of the persistent dysregulated miRNAs. In the context that many of these persistent smoking-dependent miRNAs are associated with differentiation, inflammatory diseases or lung cancer, it is likely that persistent smoking-related changes in SAE miRNAs play a role in the subsequent development of these disorders.

Show MeSH
Related in: MedlinePlus