Development and validation of exhaled breath condensate microRNAs to identify and endotype asthma in children

Autoři: Francisca Castro Mendes aff001;  Inês Paciência aff001;  António Carlos Ferreira aff004;  Carla Martins aff008;  João Cavaleiro Rufo aff001;  Diana Silva aff001;  Pedro Cunha aff009;  Mariana Farraia aff002;  Pedro Moreira aff002;  Luís Delgado aff001;  Miguel Luz Soares aff004;  André Moreira aff001
Působiště autorů: Serviço de Imunologia Básica e Clínica, Departamento de Patologia, Faculdade de Medicina da Universidade do Porto, Porto, Portugal aff001;  EPIUnit—Instituto de Saúde Pública, Universidade do Porto, Porto, Portugal aff002;  Institute of Science and Innovation in Mechanical Engineering and Industrial Management (INEGI), Porto, Portugal aff003;  Laboratório de Apoio à Investigação em Medicina Molecular (LAIMM), Faculdade de Medicina da Universidade do Porto, Porto, Portugal aff004;  Departamento de Biomedicina-Unidade de Biologia Experimental, Centro de Investigação Médica (CIM), Faculdade de Medicina da Universidade do Porto, Porto, Portugal aff005;  Pain Group, Instituto de Biologia Molecular e Celular (IBMC), Porto, Portugal aff006;  Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal aff007;  Serviço de Imunoalergologia, Centro Hospitalar São João, Porto, Portugal aff008;  Faculdade de Ciências da Nutrição e Alimentação da Universidade do Porto, Porto, Portugal aff009
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0224983


Detection and quantification of microRNAs (miRNAs) in exhaled breath condensate (EBC) has been poorly explored. Therefore we aimed to assess miRNAs in EBC as potential biomarkers to diagnose and endotype asthma in school aged children. In a cross sectional, nested case control study, all the asthmatic children (n = 71) and a random sample of controls (n = 115), aged 7 to 12 years, attending 71 classrooms from 20 local schools were selected and arbitrarily allocated to the development or validation set. Participants underwent skin-prick testing, spirometry with bronchodilation, had exhaled level of nitric oxide determined and EBC collected. Based on previous studies eleven miRNAs were chosen and analyzed in EBC by reverse transcription-quantitative real-time PCR. Principal component analysis was applied to identify miRNAs profiles and associations were estimated using regression models. In the development set (n = 89) two clusters of miRNAs were identified. After adjustments, cluster 1 and three of its clustered miRNAs, miR-126-3p, miR-133a-3p and miR-145-5p were positively associated with asthma. Moreover miR-21-5p was negatively associated with symptomatic asthma and positively associated with positive bronchodilation without symptoms. An association was also found between miR-126-3p, cluster 2 and one of its clustered miRNA, miR-146-5p, with higher FEF25-75 reversibility. These findings were confirmed in the validation set (n = 97) where two identical clusters of miRNAs were identified. Additional significant associations were observed between miR-155-5p with symptomatic asthma, negative bronchodilation with symptoms and positive bronchodilation without symptoms. We showed that microRNAs can be measured in EBC of children and may be used as potential biomarkers of asthma, assisting asthma endotype establishment.

Klíčová slova:

Asthma – Biomarkers – Diagnostic medicine – MicroRNAs – Pediatrics – Phenotypes – Pulmonary function – RNA extraction


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2019 Číslo 11