Classificação dos fenótipos da asma por análise proteômica e metabolômica

Abstract

Asthma is a heterogeneous disease characterized by chronic airway inflammation. The drug response may differ due to the inflammatory response triggered by the immune cells present in the pulmonary microenvironment. Thus, asthma phenotyping based on local inflammatory profile may aid in treatment definition and in the identification of new therapeutic targets. This study aimed to identify the changes in protein and metabolic profile of induced sputum and serum from patients with eosinophilic, neutrophilic, mixed and paucigranulocytic asthma, and to establish a panel of molecules capable of discriminating the analyzed groups. Proteomic analysis was performed by a high performance ultra-liquid chromatography (ultra-HPLC) system coupled to the quadupole Exactive Orbitrap mass spectrometer. The metabolomic profile was analyzed in an HPLC system coupled to a quadrupole-flight-time mass spectrometer. 52 proteins showed significant difference between at least 2 groups in induced sputum, while only 12 were altered between at least two groups in the patients' serum (p <0.05). Most proteins were associated with immune system pathways, mainly related to neutrophil degranulation. A panel of 5 proteins identified in induced sputum (BPIFA1, MUC5AC, CAMP, CTSG. e ANXA1) was capable of discriminating the phenotypic groups, and a panel of 4 proteins identified in serum (FCN3, THBS1, LAMP2 e CRISP3) just did not differentiate neutrophils from paucigranulocytes. In the metabolic profile, 17 metabolites that showed some significant change between the groups were identified in sputum and 15 in serum (p <0.05). Also, using the Random Forest classification algorithm, it was possible to discriminate the phenotypic groups with accuracy of 98.62% in sputum and 97.98% in serum. This study demonstrated that, due to the protein and metabolic profile of sputum and serum, it was possible to identify the molecular changes of the inflammatory phenotype of asthma. These proteins and metabolites must be validated so that these molecules can be used in asthma phenotyping, relevant to the treatment of the disease.