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Modification of gene expression of the small airway epithelium in response to cigarette smoking

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An Erratum to this article was published on 29 May 2008

Abstract

The earliest morphologic evidence of changes in the airways associated with chronic cigarette smoking is in the small airways. To help understand how smoking modifies small airway structure and function, we developed a strategy using fiberoptic bronchoscopy and brushing to sample the human small airway (10th–12th order) bronchial epithelium to assess gene expression (Affymetrix HG-U133A and HG-133 Plus 2.0 array) in phenotypically normal smokers (n = 16, 25 ± 7 pack-years) compared to matched nonsmokers (n = 17). Compared to samples from large (second to third order) bronchi, the small airway samples had a higher proportion of ciliated cells, but less basal, undifferentiated, and secretory cells, and contained Clara cells. Even though the smokers were phenotypically normal, microarray analysis of gene expression of the small airway epithelium of the smokers compared to the nonsmokers demonstrated up- and downregulation of genes in multiple categories relevant to the pathogenesis of chronic obstructive lung disease (COPD), including genes coding for cytokines/innate immunity, apoptosis, mucin, response to oxidants and xenobiotics, and general cellular processes. In the context that COPD starts in the small airways, these gene expression changes in the small airway epithelium in phenotypically normal smokers are candidates for the development of therapeutic strategies to prevent the onset of COPD.

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Acknowledgements

We thank the Pulmonary Fellows and the nurses of the bronchoscopy suite of the Division of Pulmonary and Critical Care Medicine for helping with bronchoscopies; Igor Dolgalev and Tina Raman for excellent technical assistance; T. O’Connor and N. Hackett for helpful discussions; and N. Mohamed for help in preparing this manuscript. These studies were supported, in part, by NIH R01 HL074326 and Weill Cornell GCRC M01RR00047 and the Will Rogers Memorial Fund, Los Angeles, CA.

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Authors and Affiliations

  1. Division of Pulmonary and Critical Care Medicine, Weill Medical College of Cornell University, New York, NY, USA

    Ben-Gary Harvey & Ronald G. Crystal

  2. Department of Genetic Medicine, Weill Medical College of Cornell University, 515 East 71st Street, S-1000, New York, NY, 10021, USA

    Adriana Heguy, Philip L. Leopold, Brendan J. Carolan, Barbara Ferris & Ronald G. Crystal

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  1. Ben-Gary Harvey
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  2. Adriana Heguy
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  3. Philip L. Leopold
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  4. Brendan J. Carolan
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  5. Barbara Ferris
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  6. Ronald G. Crystal
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Corresponding author

Correspondence to Ronald G. Crystal.

Additional information

Ben-Gary Harvey and Adriana Heguy contributed equally to this study.

An erratum to this article can be found at http://dx.doi.org/10.1007/s00109-008-0351-1

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Supplemental Fig. 1

Fluoroscopy of brushing to obtain large and small airway epithelial cells through a fiberoptic bronchoscope. a Proximal brush, large airway (third order), and right lower lobe. b. Distal brush, small airway (10th to 12th order), and right lower lobe. Arrows mark the tip of the brush(PDF 476 kb)

Supplemental Fig. 2

Hierarchical cluster analysis of airway epithelial gene expression changes in the small airways of healthy smokers vs nonsmokers. a Dendrogram resulting from cluster analysis for all expressed genes (30,963 probe sets, present in at least one array), using MAS5-generated values and the GeneSpring software (Spearman correlation) by sample. Note that this analysis does not segregate smokers from nonsmokers, indicating that cigarette smoking does not change the global pattern of gene expression in the epithelium of the small airways. b. Cluster analysis for genes differentially expressed in smokers vs nonsmokers. The cluster was generated for 118 significant probe sets [p < 0.05, Welch t test with the Benjamini–Hochberg false discovery rate multiple test correction, by both MAS5 and RMA preprocessing algorithms; >1.5-fold change in smokers vs nonsmokers (upregulated) or nonsmokers vs smokers (downregulated), calculated by either the MAS5 values or the RMA values], using the GeneSpring program (Spearman correlation) by sample, and by gene. Smokers and nonsmokers were not grouped a priori. Genes expressed above average are represented in red, genes expressed below average in blue, and average levels in white. The degree of red or blue intensity indicates the degree of up- or downregulation relative to the average level of expression across all samples. The genes are represented vertically, and the individual samples [S for smokers (n = 10) and NS for nonsmokers (n = 12)] horizontally(PDF 218 kb)

Supplemental Table 1

Demographics of the Study Population1 (PDF 91 kb).

Supplemental Table 2

Genes Differentially Expressed in the Small Airway Epithelium of 6 Healthy Smokers vs 5 Healthy Non-smokers (PDF 82 kb).

Supplemental Table 3

Examples of Genes Differentially Expressed in the Small Airway Epithelium in Smokers vs Non-smokers from Group A in Functional Categories that are Relevant for the Pathogenesis of COPD (PDF 40 kb).

Supplemental Table 4

Genes Differentially Expressed in the Small Airway Epithelium of 10 Healthy Smokers vs 12 Healthy Non-smokers (PDF 155 kb).

Supplemental Table 5

Examples of Genes Differentially Expressed in the Small Airway Epithelium of 10 Smokers vs 12 Non-smokers, in Functional Categories that are Relevant for the Pathogenesis of COPD1 (PDF 34 kb).

Supplemental Table 6

Comparison of HG-U133 2.0 vs. HG-U133A results (PDF 104 kb).

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Harvey, BG., Heguy, A., Leopold, P.L. et al. Modification of gene expression of the small airway epithelium in response to cigarette smoking. J Mol Med 85, 39–53 (2007). https://doi.org/10.1007/s00109-006-0103-z

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