Michael J. McGeachie, PhD, Joshua S. Davis, MD, Alvin T

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Asthma remission: Predicting future airways responsiveness using an miRNA network Michael J. McGeachie, PhD, Joshua S. Davis, MD, Alvin T. Kho, PhD, Amber Dahlin, PhD, Joanne E. Sordillo, ScD, Maoyun Sun, PhD, Quan Lu, PhD, Scott T. Weiss, MD, Kelan G. Tantisira, MD Journal of Allergy and Clinical Immunology Volume 140, Issue 2, Pages 598-600.e8 (August 2017) DOI: 10.1016/j.jaci.2017.01.023 Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 A, Markov neighborhood of conditional Gaussian BN predictive of AHR remission at 14 years. Included are 11 miRs and baseline PC20. Arrows indicate statistical dependence of the target node on the source node. Arrow thickness indicates strength of connection in log Bayes factor. Node color and size indicate the number of connections. See this article's Methods section in the Online Repository at www.jacionline.org and McGeachie et al3 for additional details. B, Receiver operating characteristic curve and convex hull (ROCCH) for prediction of 14-year AHR remission using Bayesian in Fig 1, A. The convex hull of raw receiver operating characteristic curves represents achievable tradeoffs between false positives and false negatives using the classifiers. Area under the convex hull of the curve (AUC) is 86.5% for the Bayesian model including baseline PC20 and 80.0% for the Bayesian model of just miRs. Journal of Allergy and Clinical Immunology 2017 140, 598-600.e8DOI: (10.1016/j.jaci.2017.01.023) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Number of subjects from the CAMP study who had microRNA data available and had AHR remission, as quantified by a PC20 value above the limit of 37.5 mg/mL in a methacholine challenge. The x-axis is displayed in months since randomization in the CAMP trial. Top panel: There was a noticeable peak at 168 months (14 years) from baseline of subjects in AHR remission. Journal of Allergy and Clinical Immunology 2017 140, 598-600.e8DOI: (10.1016/j.jaci.2017.01.023) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Empirical distribution of permutation test realizations for BN predicting 14-year AHR remission. For each of 1000 iterations, the phenotype labels were shuffled (AHR remission) and a new BN was learned from the miRs to predict the scrambled phenotype. For each iteration, the BN's prediction on the scrambled phenotype is reported (AUC), and shown in the histogram above. The BN model on the true data exhibits statistically significant prediction (86% AUC; P = .006). Journal of Allergy and Clinical Immunology 2017 140, 598-600.e8DOI: (10.1016/j.jaci.2017.01.023) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Conditional Gaussian BN predictive of AHR remission at 14 years. Arrows indicate statistical dependence of the target node on the source node. Arrow thickness indicates the strength of connection in log Bayes factor. Node color and size indicate the number of connections. Journal of Allergy and Clinical Immunology 2017 140, 598-600.e8DOI: (10.1016/j.jaci.2017.01.023) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Quantile-Quantile plot of logistic regression tests for each miR's association with 14-year AHR remission. This indicates that no miRs are significantly associated with AHR remission after correction for multiple testing. GIF, Genomic inflation factor. Journal of Allergy and Clinical Immunology 2017 140, 598-600.e8DOI: (10.1016/j.jaci.2017.01.023) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E5 HASM cells were transfected with 10 nM of either scramble control or miR-30a/d/e mimics. Seventy-two hours after transfection, cells were trypsinized for 8 minutes and measured for cell size by the Moxi Z Cell Analyzer. Average cell diameter (μm) was compared in mimic-transfected versus scramble-transfected HASM cells. Data (mean ± SE) were obtained from 3 independent experiments. ctrl, Control samples; scr, scramble-transfected control samples. *Statistically significant increases in cell diameter (hypertrophy) over scramble cells at P < .05. Journal of Allergy and Clinical Immunology 2017 140, 598-600.e8DOI: (10.1016/j.jaci.2017.01.023) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions