Katherine J. Baines, PhD, Jodie L. Simpson, PhD, Lisa G

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Sputum gene expression signature of 6 biomarkers discriminates asthma inflammatory phenotypes Katherine J. Baines, PhD, Jodie L. Simpson, PhD, Lisa G. Wood, PhD, Rodney J. Scott, PhD, Naomi L. Fibbens, DipPathTech, Heather Powell, MMedSci, Douglas C. Cowan, PhD, D. Robin Taylor, DSc, Jan O. Cowan, Peter G. Gibson, MBBS Journal of Allergy and Clinical Immunology Volume 133, Issue 4, Pages 997-1007 (April 2014) DOI: 10.1016/j.jaci.2013.12.1091 Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Schematic diagram documenting the biomarker identification and evaluation process undertaken. AUS, Australian population; NZ, New Zealand population; ROC, receiver operating characteristic; 6GS, 6-gene expression signature. Journal of Allergy and Clinical Immunology 2014 133, 997-1007DOI: (10.1016/j.jaci.2013.12.1091) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Relative gene expression levels of CLC (A), CPA3 (B), DNASE1L3 (C), IL1B (D), ALPL (E), and CXCR2 (F) in induced sputum samples from patients with inflammatory phenotypes of asthma compared with healthy control subjects. Gene expression is calculated relative to that of β-actin and expressed as a fold change from the healthy control group mean. Bar graphs show means ± SEMs. *P < .05 versus healthy control subjects, #P < .05 versus PGA, ˆP < .05 versus EA, and ∼P < .05 versus NA. Journal of Allergy and Clinical Immunology 2014 133, 997-1007DOI: (10.1016/j.jaci.2013.12.1091) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Receiver operating characteristic curves demonstrate that the 6-gene biomarker signature identifies ICS treatment responsiveness determined by a 12% change in FEV1 (A), a 0.5-point decrease in ACQ score (B), and a 40% reduction in Feno value (C). AUC, Area under the curve. Journal of Allergy and Clinical Immunology 2014 133, 997-1007DOI: (10.1016/j.jaci.2013.12.1091) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Relative gene expression levels of CLC (A), CPA3 (B), DNASE1L3 (C), IL1B (D), ALPL (E), and CXCR2 (F) in induced sputum samples from subjects with EA or NEA before and after corticosteroid treatment. Gene expression is calculated relative to that of β-actin and expressed as a fold change from the healthy control group mean. *P < .05 versus healthy control subjects, #P < .05 versus patients with NEA before ICSs. Journal of Allergy and Clinical Immunology 2014 133, 997-1007DOI: (10.1016/j.jaci.2013.12.1091) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E1 Journal of Allergy and Clinical Immunology 2014 133, 997-1007DOI: (10.1016/j.jaci.2013.12.1091) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E2 Journal of Allergy and Clinical Immunology 2014 133, 997-1007DOI: (10.1016/j.jaci.2013.12.1091) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E3 Journal of Allergy and Clinical Immunology 2014 133, 997-1007DOI: (10.1016/j.jaci.2013.12.1091) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E4 Journal of Allergy and Clinical Immunology 2014 133, 997-1007DOI: (10.1016/j.jaci.2013.12.1091) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig E5 Journal of Allergy and Clinical Immunology 2014 133, 997-1007DOI: (10.1016/j.jaci.2013.12.1091) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions