Waterpipe smoking induces epigenetic changes in the small airway epithelium

PLoS One. 2017 Mar 8;12(3):e0171112. doi: 10.1371/journal.pone.0171112. eCollection 2017.


Waterpipe (also called hookah, shisha, or narghile) smoking is a common form of tobacco use in the Middle East. Its use is becoming more prevalent in Western societies, especially among young adults as an alternative form of tobacco use to traditional cigarettes. While the risk to cigarette smoking is well documented, the risk to waterpipe smoking is not well defined with limited information on its health impact at the epidemiologic, clinical and biologic levels with respect to lung disease. Based on the knowledge that airway epithelial cell DNA methylation is modified in response to cigarette smoke and in cigarette smoking-related lung diseases, we assessed the impact of light-use waterpipe smoking on DNA methylation of the small airway epithelium (SAE) and whether changes in methylation were linked to the transcriptional output of the cells. Small airway epithelium was obtained from 7 nonsmokers and 7 light-use (2.6 ± 1.7 sessions/wk) waterpipe-only smokers. Genome-wide comparison of SAE DNA methylation of waterpipe smokers to nonsmokers identified 727 probesets differentially methylated (fold-change >1.5, p<0.05) representing 673 unique genes. Dominant pathways associated with these epigenetic changes include those linked to G-protein coupled receptor signaling, aryl hydrocarbon receptor signaling and xenobiotic metabolism signaling, all of which have been associated with cigarette smoking and lung disease. Of the genes differentially methylated, 11.3% exhibited a corresponding significant (p<0.05) change in gene expression with enrichment in pathways related to regulation of mRNA translation and protein synthesis (eIF2 signaling and regulation of eIF4 and p70S6K signaling). Overall, these data demonstrate that light-use waterpipe smoking is associated with epigenetic changes and related transcriptional modifications in the SAE, the cell population demonstrating the earliest pathologic abnormalities associated with chronic cigarette smoking.

MeSH terms

  • Adult
  • Bronchi / metabolism
  • DNA / genetics
  • DNA / isolation & purification
  • DNA / metabolism
  • DNA Methylation
  • Down-Regulation
  • Epigenesis, Genetic*
  • Epithelium / metabolism*
  • Female
  • Genome, Human
  • Humans
  • Male
  • RNA / genetics
  • RNA / isolation & purification
  • RNA / metabolism
  • Receptors, Aryl Hydrocarbon / metabolism
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism
  • Smoking*
  • Up-Regulation


  • Receptors, Aryl Hydrocarbon
  • Receptors, G-Protein-Coupled
  • RNA
  • DNA

Grants and funding

This work was supported by the National Heart, Lung, and Blood Institute through grants HL107882, HL107882-2S1 and P20 HL113443 to RGC, by the National Institutes of Health through grants UL1 TR000457 and RR024143 and by the Qatar National Research Fund through grant NPRP 09-742-3-194 to MSW. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.