Purpose: To identify DNA methylation-based biomarkers predictive of outcome in locally advanced non-small cell lung cancer (LA-NSCLC) patients undergoing definitive chemoradiotherapy. Methods: Genome-wide DNA methylation profiling was performed on pre-treatment tumor biopsies from 89 LA-NSCLC patients. Differentially methylated regions were identified and a predictive signature was developed. Results: A 12-CpG methylation signature significantly stratified patients by progression-free survival (HR 2.4, p<0.001). The signature was validated in an independent cohort. Conclusions: DNA methylation profiling identifies LA-NSCLC patients at high risk of treatment failure who may benefit from intensified or alternative treatment strategies.