An Integrated Prognostic Classifier for Stage I Lung Adenocarcinoma Based on mRNA, microRNA, and DNA Methylation Biomarkers Ana I. Robles, PhD, Eri Arai,

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An Integrated Prognostic Classifier for Stage I Lung Adenocarcinoma Based on mRNA, microRNA, and DNA Methylation Biomarkers Ana I. Robles, PhD, Eri Arai, MD, Ewy A. Mathé, PhD, Hirokazu Okayama, MD, PhD, Aaron J. Schetter, PhD, MPH, Derek Brown, MS, David Petersen, BS, Elise D. Bowman, MS, Rintaro Noro, MD, Judith A. Welsh, BS, Daniel C. Edelman, PhD, Holly S. Stevenson, PhD, Yonghong Wang, PhD, Naoto Tsuchiya, PhD, Takashi Kohno, PhD, Vidar Skaug, MD, Steen Mollerup, MD, Aage Haugen, MD, Paul S. Meltzer, MD, PhD, Jun Yokota, MD, Yae Kanai, MD, PhD, Curtis C. Harris, MD Journal of Thoracic Oncology Volume 10, Issue 7, Pages 1037-1048 (July 2015) DOI: 10.1097/JTO.0000000000000560 Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Methylation of genes marked by H3K27me3 in embryonic stem cells (ESC) is associated with clinical outcome. Hierarchical clustering based on hypermethylated CpGs of genes marked by H3K27me3 in ESC in the National Cancer Institute (NCI; A) and Japan (C) microarray cohorts. Each column represents an individual patient and each row an individual CpG probe. Patients in the high methylation clusters had shorter cancer-specific (B) or relapse-free (D) survival in Kaplan–Meier survival analysis. Journal of Thoracic Oncology 2015 10, 1037-1048DOI: (10.1097/JTO.0000000000000560) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Kaplan–Meier analysis of HOXA9 promoter methylation in stage I lung adenocarcinoma (ADC). HOXA9 methylation values were dichotomized based on ≥40% or <40% mean methylation in pyrosequencing analysis. The associations were evaluated in two independent cohorts, including the combined National Cancer Institute (NCI)/Norway cohort (left panels) and the Japan cohort (right panels). Shown are the analysis of stage I (A) and the subgroup analysis of stage IA (B) and stage IB (C). The p values were calculated by log-rank test. Journal of Thoracic Oncology 2015 10, 1037-1048DOI: (10.1097/JTO.0000000000000560) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Kaplan–Meier analysis of a combined prognostic biomarker in stage I lung adenocarcinoma (ADC). Patients in the combined National Cancer Institute (NCI)/Norway cohort (left panels) and the Japan cohort (right panels) were categorized according to the combined number of high values for HOXA9 methylation, miR-21, and 4-protein-coding gene signature. An increasing combined score conferred greater risk for poor outcome in stage I (A), and within subgroup analysis of stage IA (B) and stage IB (C). The p values were calculated by log-rank test. Journal of Thoracic Oncology 2015 10, 1037-1048DOI: (10.1097/JTO.0000000000000560) Copyright © 2015 International Association for the Study of Lung Cancer Terms and Conditions