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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.

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Effect of smoking cessation on the SAE expression of the 34 miRNAs dysregulated by smoking.A: Quantitative evaluation of smoking cessation. Urine nicotine and cotinine levels of healthy nonsmokers, healthy smoker before smoking cessation (month 0), healthy smokers after 3 months smoking cessation. B: Global effect of smoking cessation on smoking-dependent miRNAs. Smoking dependent miRNA indexes based on 34 smoking dependent miRNAs were calculated (see Methods) for each of the healthy nonsmokers, healthy smokers before smoking cessation (month 0) and healthy smokers after 3 months smoking cessation. C: Volcano plot of the 34 smoking-dependent miRNAs in healthy smokers after 3-month smoking cessation compared to healthy nonsmokers. X-axis, fold-change, healthy smokers after 3-month smoking cessation vs healthy nonsmokers; Y-axis, p value. Twelve of the 34 miRNAs were not reversed after smoking cessation (criteria p<0.05, fold-change >1.5). D: Quantitative assessment of smoking cessation on miRNA expression in human SAE. Using mean absolute expression levels, the degree of reversibility in a miRNA was calculated as: % change = [(healthy smokers before smoking cessation—healthy smokers after 3-month smoking cessation)x100 / (healthy smokers before smoking cessation—healthy nonsmokers)]. X-axis—percentage of the smoking dependent miRNA (in order of ascending % change). The persistent miRNA identified in panel C are shown as red. Y-axis—% change of miRNA. The vertical dash line indicates the percentage of miRNAs which have 25% and 50% reversal.
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pone.0120824.g002: Effect of smoking cessation on the SAE expression of the 34 miRNAs dysregulated by smoking.A: Quantitative evaluation of smoking cessation. Urine nicotine and cotinine levels of healthy nonsmokers, healthy smoker before smoking cessation (month 0), healthy smokers after 3 months smoking cessation. B: Global effect of smoking cessation on smoking-dependent miRNAs. Smoking dependent miRNA indexes based on 34 smoking dependent miRNAs were calculated (see Methods) for each of the healthy nonsmokers, healthy smokers before smoking cessation (month 0) and healthy smokers after 3 months smoking cessation. C: Volcano plot of the 34 smoking-dependent miRNAs in healthy smokers after 3-month smoking cessation compared to healthy nonsmokers. X-axis, fold-change, healthy smokers after 3-month smoking cessation vs healthy nonsmokers; Y-axis, p value. Twelve of the 34 miRNAs were not reversed after smoking cessation (criteria p<0.05, fold-change >1.5). D: Quantitative assessment of smoking cessation on miRNA expression in human SAE. Using mean absolute expression levels, the degree of reversibility in a miRNA was calculated as: % change = [(healthy smokers before smoking cessation—healthy smokers after 3-month smoking cessation)x100 / (healthy smokers before smoking cessation—healthy nonsmokers)]. X-axis—percentage of the smoking dependent miRNA (in order of ascending % change). The persistent miRNA identified in panel C are shown as red. Y-axis—% change of miRNA. The vertical dash line indicates the percentage of miRNAs which have 25% and 50% reversal.

Mentions: Assessment of urine nicotine and cotinine confirmed that the 10 smokers had quit, with urine nicotine and cotinine measured in the nonsmoking range at the 3 month assessment (Fig 2A). To assess the effect of smoking cessation on the expression of smoking-dependent miRNAs at a global level, a smoking-dependent miRNA index was used. Smoking up-regulated the smoking-dependent miRNA index (3.8-fold, p<0.05) and smoking cessation was associated with significant down-regulation of the index (1.6-fold, p<0.05, compared to healthy smokers, Fig 2B). When assessed at this global level of all smoking changed miRNAs, the differences between healthy nonsmokers and smokers that quit was not statistically significant, although the average miRNA index of the smoking cessation group was still 2.4-fold higher than that of the healthy nonsmokers, suggesting the levels of individual miRNAs may still be significantly abnormal even with smoking cessation.


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)

Effect of smoking cessation on the SAE expression of the 34 miRNAs dysregulated by smoking.A: Quantitative evaluation of smoking cessation. Urine nicotine and cotinine levels of healthy nonsmokers, healthy smoker before smoking cessation (month 0), healthy smokers after 3 months smoking cessation. B: Global effect of smoking cessation on smoking-dependent miRNAs. Smoking dependent miRNA indexes based on 34 smoking dependent miRNAs were calculated (see Methods) for each of the healthy nonsmokers, healthy smokers before smoking cessation (month 0) and healthy smokers after 3 months smoking cessation. C: Volcano plot of the 34 smoking-dependent miRNAs in healthy smokers after 3-month smoking cessation compared to healthy nonsmokers. X-axis, fold-change, healthy smokers after 3-month smoking cessation vs healthy nonsmokers; Y-axis, p value. Twelve of the 34 miRNAs were not reversed after smoking cessation (criteria p<0.05, fold-change >1.5). D: Quantitative assessment of smoking cessation on miRNA expression in human SAE. Using mean absolute expression levels, the degree of reversibility in a miRNA was calculated as: % change = [(healthy smokers before smoking cessation—healthy smokers after 3-month smoking cessation)x100 / (healthy smokers before smoking cessation—healthy nonsmokers)]. X-axis—percentage of the smoking dependent miRNA (in order of ascending % change). The persistent miRNA identified in panel C are shown as red. Y-axis—% change of miRNA. The vertical dash line indicates the percentage of miRNAs which have 25% and 50% reversal.
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pone.0120824.g002: Effect of smoking cessation on the SAE expression of the 34 miRNAs dysregulated by smoking.A: Quantitative evaluation of smoking cessation. Urine nicotine and cotinine levels of healthy nonsmokers, healthy smoker before smoking cessation (month 0), healthy smokers after 3 months smoking cessation. B: Global effect of smoking cessation on smoking-dependent miRNAs. Smoking dependent miRNA indexes based on 34 smoking dependent miRNAs were calculated (see Methods) for each of the healthy nonsmokers, healthy smokers before smoking cessation (month 0) and healthy smokers after 3 months smoking cessation. C: Volcano plot of the 34 smoking-dependent miRNAs in healthy smokers after 3-month smoking cessation compared to healthy nonsmokers. X-axis, fold-change, healthy smokers after 3-month smoking cessation vs healthy nonsmokers; Y-axis, p value. Twelve of the 34 miRNAs were not reversed after smoking cessation (criteria p<0.05, fold-change >1.5). D: Quantitative assessment of smoking cessation on miRNA expression in human SAE. Using mean absolute expression levels, the degree of reversibility in a miRNA was calculated as: % change = [(healthy smokers before smoking cessation—healthy smokers after 3-month smoking cessation)x100 / (healthy smokers before smoking cessation—healthy nonsmokers)]. X-axis—percentage of the smoking dependent miRNA (in order of ascending % change). The persistent miRNA identified in panel C are shown as red. Y-axis—% change of miRNA. The vertical dash line indicates the percentage of miRNAs which have 25% and 50% reversal.
Mentions: Assessment of urine nicotine and cotinine confirmed that the 10 smokers had quit, with urine nicotine and cotinine measured in the nonsmoking range at the 3 month assessment (Fig 2A). To assess the effect of smoking cessation on the expression of smoking-dependent miRNAs at a global level, a smoking-dependent miRNA index was used. Smoking up-regulated the smoking-dependent miRNA index (3.8-fold, p<0.05) and smoking cessation was associated with significant down-regulation of the index (1.6-fold, p<0.05, compared to healthy smokers, Fig 2B). When assessed at this global level of all smoking changed miRNAs, the differences between healthy nonsmokers and smokers that quit was not statistically significant, although the average miRNA index of the smoking cessation group was still 2.4-fold higher than that of the healthy nonsmokers, suggesting the levels of individual miRNAs may still be significantly abnormal even with smoking cessation.

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