Dra Amanda Iglesias, DUE Rocio Cordova, Hospital Universitario Son Espases-IdISPa,

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1 On-line data supplement Authorship Members of the CHACOS study group Dra Amanda Iglesias, DUE Rocio Cordova, Hospital Universitario Son Espases-IdISPa, CIBERES, Palma de Mallorca, Spain; Dr. Sergi Pascual, Dra. Mireia Admetlló, Hospital del Mar, Barcelona, Spain; Dra. Salud Santos, Dr. Gianluca Cotta Ramusino, Hospital Universitari de Bellvitge, Barcelona, Spain; Dra. Carmen Gómez Neira, Dra. Irene Carla Martin Robles, Hospital Universitario Lucus Augusti, Lugo, Spain; Dr. Alberto Fernández-Villar, Dra. Cristina Represas, Dra. Ana Priegue Carreracon Hospital Xeneral de Vigo, Vigo, Spain; Dra. Andrea Trisán Alonso Hospital Puerta del Hierro, Madrid, Spain; Dr. Francisco García Río, Dr. Raúl Galera Martínez, Dra. Raquel Casitas Mateo, Dra. Elisabet Martínez Cerón, Hospital La Paz, Madrid, Spain; Dra. Sagrario Mayoralas, Dr. Adalberto Pacheco Galván, Hospital Universitario Ramón y Cajal, Madrid, Spain; Dr. Luis Puente Maestu, Dra. Julia Garcia de Pedro, Dra. Pilar Sanz, Hospital General Universitario Gregorio Marañón, Madrid, Spain; Dr. José Belda Ramírez, Dr. Fernando Sánchez-Toril López, Dr. Rafael Peris Cardells Hospital Arnau de Vilanova, Valencia, Spain; Dra. Maria Climent Gregori, Hospital Universitario Dr. Peset, Valencia, Spain; Dra. Patricia Garcia-Sidro, Hospital Castellón de la Plana; Dra. Cleofe Fernández, Dra. Mª Encarnación Barroso Medel, Hospital General Universitario de Alicante, Alicante, Spain; Dra. Carmen Calero Acuña, Dra. María Abad Arranz, Dr. Eduardo Márquez Martín, Dr. Francisco Ortega Ruiz Hospital Virgen del Rocío, Sevilla, Spain; Dr. Bernardino Alcazar, Hospital Alta Resolución de la Loja, Granada, Spain; Dr. Raúl Godoy Mayoral, Javier Cruz Ruiz, Hospital General Universitario de Albacete, Albacete, Spain; Dra. Nuria Marina Malanda, Dr. Juan Bautista Gáldiz Iturri, Dra. Elena López de Santa Maria Miró, Hospital Universitario

2 Cruces, Bilbao, Spain; Dr. Juan Luis García Rivero, Dra. Ruth Valerie Alain Mbessekg, Hospital de Laredo, Laredo, Spain; Dra. Cristina Martinez, Dra. Rosirys Mercedes Guzmán Taveras, Dra. Ana Fernandez Tena, Dra. Rosario Madiedo de la Llera, Dra. Ángeles Encarnación Malmierca de Dios Instituto Nacional de Silicosis, Oviedo, Spain; Dr. Jose Antonio Ros Lucas, Dra. M. Carmen Soto Fernández, Dr. Rubén Andújar Espinosa, Hospital Universitario Virgen de la Arrixaca, Murcia, Spain; Dra. Míriam Barrecheguren, Dra. Cristina Esquinas Hospital Universitari Vall d'hebrón, Barcelona, Spain Material and methods Definitions -Asthma was diagnosed -according to international guidelines (11)- by the presence of symptoms of wheeze, breathlessness, or cough plus a positive bronchodilator test (significant improvement by >12% and 200 ml in the forced expiratory volume in 1 s - FEV1-10 min after the inhalation of salbutamol) or a positive methacholine test (a provocative concentration of methacholine required to lower the FEV1 by 20% -PC20- of <4 mg/ml). -COPD was diagnosed according to international recommendations (2)- by the presence of post-bronchodilator FEV1/FVC <0.70 in patients with smoking history of > 10 pack-years.

3 Study protocol All researchers were asked to recruit 12 consecutive patients with COAD: 4 NSA, 4 COPD, 2 SA and 2 COPD-e (Figure 1). The recruitment period was between October 2015 and March The study was conducted in a single visit in which the researchers obtained and recorded in an electronic clinical research database (ecrd): anthropometric data, smoking history, previous history of atopy, rhinitis, nasal polyposis and comorbidities, exacerbations in the previous year, symptoms assessed by the validated Spanish version of the COPD Assesment Test (CAT) and by the Asthma Control Test (ACT), pharmacologic treatments and the degree of compliance with inhalers estimated by the Test of Adherence to Inhalers (TAI) (16). Fractional exhaled nitric oxide (FeNO) was measured by a chemolumines-cence analyser (NIOX, Aerocrine, Stockholm, Sweden) at a flow rate of 50 ml/s, in accordance with the recommendations of the ATS/ERS task force (19). Sputum induction Before induction of sputum, patients inhaled 400 μg of salbutamol. Briefly, the volume of the whole sputum was assessed and an equal volume of dithioerythritol (Sigma, Munich, Germany) was added. The total cell count and viability were obtained simultaneously in a modified Neubauer counting chamber (Hausser Scientific, Horsham, PA, USA). Differential cell counts were expressed as the percentage of nonsquamous cells and those with significant squamous contamination (> 80%) were excluded from analysis.

4 Sample size calculation An a priori sample size was calculated considering a statistical power of 80% and 5% of alpha error. A minimum of 100 ACOS patients and 200 other chronic airways disease patients would be enough in order to identify statistically significant differences for differences of 15% or higher when compared with any of the reference groups, in most variables ranging from 10% up to 80% in ACOS.

5 Results Table S1. Characteristics of patients according to the baseline diagnosis with corrected post hoc 2 group comparisons. Number of subjects Age, years Gender (% female) Total Sample (10.9) 38.4 Non-smoking asthmatics (A) COPD (B) ACOS (C) (12.3) 67.9 (8.6) p value overall 63.3 (10.6) < <0.001 p-value A vs B p-value B vs C p-value A vs C < < BMI (Kg/m (5.2) 28.4 ) 28.6 (4.7) (4.9) 28.5 (6) 0.96 Pack/year 32.7 (28.4) (5.8) (22.5) 45.0 (24.9) <0.001 < <0.001 Age of onset (yrs) 47.5 (18.9) 58.9 <0.001 <0.001 < (18.6) 48.5 (19.1) <0.001 (8.7) Positive SPT (%) <0.001 <0.001 < Rhinitis (%) <0.001 < Patients with nocturnal symptoms (%) Comorbidities: -Arterial hypertension (%) -Diabetes (%) -Ischemic heart

6 disease (%) -Heart failure (%) -Anemia (%) -Osteoporosis (%) -Psychiatric disorders (8%) -Gastroesophageal Reflux (%) < PreBD FEV1 (%) 55.5 (17.8) (17.7) 53.7 (16.5) <0.001 (18.2) PostBD FEV1(%) 61.1 (18.9) 55.1 < (18.9) 58.9 (17.0) <0.001 (18.5) PBT (%) DLCO (%) 71.2 (22.9) 63.6 < < (18,7) 66.5 (16.1) <0.001 (25,9) Exacerbations 1.00 (1.35) (1.37) (1.32) 0.97 (1.36) 0.85 Blood Eos 205 (0, 100 <0.001 < (0, 4350) 300 (0, 1000) <0.001 (cells/µl) 4350) (0,200) Sputum 4.3 (5.1) (5.8) 1.8 (2.1) 6.3 (5.8) eosinophils <0,001 (n=35, %) IgE (IU/ml) 74 (4, 2909) (4, 2909) (4,2301) 90 (4, 2500) FeNO (ppb) 25.3 (21.1) 17.3 < < (24.1) 22.6 (25.2) <0.001 (9.6) CAT 13.6 (7.7) 14.8 (8.8) (7.7) 0.64

7 (6.7) ACT 19.9 (4.6) (4.4) (4.3) 19.4 (4.8) 0.33 ICS (%) < 0.01 < <0.001 LABA (%) LAMA (%) <0.01 < PD4I (%) LTA (%) < 0.01 < <0.001 Omalizumab (%) < Oral 2.1 corticosteroids (%) < TAI 47.8 (3.9) (3.9) (4.0) 47.6 (3.9) 0.06 Numerical data are expressed as mean (SD) except for IgE and eosinophils which are expressed as median (range) or in percentage (%); SPT: skin prick test; prebd: pre-bronchodilator FEV1; postbd: post-bronchodilator FEV1; PBT: rate of patients with positive bronchodilator test; DLCO: carbon monoxide diffusing capacity; Exacerbations: number of severe exacerbations during the past 12 months; FeNO: fractioned exhaled nitric oxide levels; CAT: COPD Assessment Test; ACT: Asthma Control Test; ICS: Inhaled corticosteroids; LABA: Long-acting β2- agonists; LAMA: Long-acting muscarinic antagonists; PD4I: Phosphodiesterase 4 inhibitors; LTA: leukotriene receptor antagonists; TAI: Test of Adherence to Inhalers score. P-values from Student s t test for differences between means and Chi-Square tests for differences between proportions.

8 Table S2. Differential characteristics between patients according to study ACOS definition (pooled SA and e-copd) or according to consensus ACOS definition Age, years Gender (%female) Study definition N=109 Consensus definition N=58 p value 63.3 (10.6) 59.1 (10.6) (34.9%) 23 (39.7) Pack-years 45.0 (24.9) 35.2 (18.0) Positive SPT (%) 35 (32.1%) 27 (46.6) Rhinitis (%) 33 (31.1%) 28 (49.1) PostBD FEV1% 57.8 (18.9) 67.3 (18.5) Blood eosinophil count (cells/ µl) (178.7) (279.3) FeNO (ppb) 22.6 (21.6) 23.4 (18.6) Exacerbations 1.0 (1.4) 0.9 (1.3) CAT 13.3 (7.7) 13.7 (8.1) ACT 19.4 (4.8) 20.0 (4.9) Numerical data are expressed as mean (SD) except for IgE and eosinophils which are expressed as median (range) or in percentage (%); SPT: skin prick test; prebd: prebronchodilator FEV1; postbd: post-bronchodilator FEV1; PBT: rate of patients with positive bronchodilator test; Exacerbations: number of severe exacerbations during the past 12 months; FeNO: fractioned exhaled nitric oxide levels; CAT: COPD Assessment Test; ACT: Asthma Control Test; p values represent Student s t test for differences between means and Chi-Square tests for differences between proportions.

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