Smoking-Associated DNA Methylation Biomarkers and Their Predictive Value for All-Cause and Cardiovascular Mortality

Abstract

Background: With epigenome-wide mapping of DNA methylation, a number of novel smoking-associated loci have been identified.

Objectives: We aimed to assess dose-response relationships of methylation at the top hits from the epigenome-wide methylation studies with smoking exposure as well as with total and cause-specific mortality.

Methods: In a population-based prospective cohort study in Germany, methylation was quantified in baseline blood DNA of 1,000 older adults by the Illumina 450K assay. Deaths were recorded during a median follow-up of 10.3 years. Dose-response relationships of smoking exposure with methylation at nine CpGs were modeled by restricted cubic spline regression. Associations of individual and aggregate methylation patterns with all-cause, cardiovascular, and cancer mortality were assessed by multiple Cox regression.

Results: Clear dose-response relationships with respect to current and lifetime smoking intensity were consistently observed for methylation at six of the nine CpGs. Seven of the nine CpGs were also associated with mortality outcomes to various extents. A methylation score based on the top two CpGs (cg05575921 and cg06126421) showed the strongest associations with all-cause, cardiovascular, and cancer mortality, with adjusted hazard ratios (95% CI) of 3.59 (2.10, 6.16), 7.41 (2.81, 19.54), and 2.48 (1.01, 6.08), respectively, for participants with methylation levels in the lowest quartile at both CpGs. Adding methylation at those two CpGs into a model that included the variables of the Systematic Coronary Risk Evaluation chart for fatal cardiovascular risk prediction improved the predictive discrimination.

Conclusion: The novel methylation biomarkers are highly informative for both smoking exposure and smoking-related mortality outcomes. In particular, these biomarkers may substantially improve cardiovascular risk prediction. Nevertheless, the findings of the present study need to be further validated in additional large longitudinal studies.

Citation: Zhang Y, Schöttker B, Florath I, Stock C, Butterbach K, Holleczek B, Mons U, Brenner H. 2016. Smoking-associated DNA methylation biomarkers and their predictive value for all-cause and cardiovascular mortality. Environ Health Perspect 124:67-74; http://dx.doi.org/10.1289/ehp.1409020.

Figure 1

Dose–response relationships between smoking behavior and methylation intensity (results from restricted cubic spline regression adjusted for potential confounding factors). CL, confidence limit. (A) Dose–response relationship between current intensity of smoking and methylation intensity at AHRR (cg05575921; left), and 6p21.33 (cg06126421; right); never and former smokers were defined as reference, with current smoking intensity = 0. (B) Dose–response relationship between cumulative dose of smoking and methylation intensity at AHRR (cg05575921; left), and 6p21.33 (cg06126421; right); never smokers were defined as reference, with pack-years = 0. (C) Dose–response relationship between time since cessation of smoking and methylation intensity at AHRR (cg05575921; left), and 6p21.33 (cg06126421; right) among former smokers; current smokers were defined as reference, with time since cessation = 0.