ABSTRACT
Recent advances in genetics have spurred rapid progress towards the systematic identification of genes involved in complex diseases. Still, the detailed understanding of the molecular and physiological mechanisms through which these genes affect disease phenotypes remains a major challenge. Here, we identify the asthma disease module, i.e. the local neighborhood of the interactome whose perturbation is associated with asthma, and validate it for functional and pathophysiological relevance, using both computational and experimental approaches. We find that the asthma disease module is enriched with modest GWAS P-values against the background of random variation, and with differentially expressed genes from normal and asthmatic fibroblast cells treated with an asthma-specific drug. The asthma module also contains immune response mechanisms that are shared with other immune-related disease modules. Further, using diverse omics (genomics, gene-expression, drug response) data, we identify the GAB1 signaling pathway as an important novel modulator in asthma. The wiring diagram of the uncovered asthma module suggests a relatively close link between GAB1 and glucocorticoids (GCs), which we experimentally validate, observing an increase in the level of GAB1 after GC treatment in BEAS-2B bronchial epithelial cells. The siRNA knockdown of GAB1 in the BEAS-2B cell line resulted in a decrease in the NFkB level, suggesting a novel regulatory path of the pro-inflammatory factor NFkB by GAB1 in asthma.
Subject(s)
Anti-Asthmatic Agents/pharmacology , Asthma/genetics , Adaptor Proteins, Signal Transducing/genetics , Adaptor Proteins, Signal Transducing/metabolism , Anti-Asthmatic Agents/therapeutic use , Asthma/drug therapy , Asthma/metabolism , Base Sequence , Dose-Response Relationship, Drug , Gene Expression , Gene Expression Regulation , Gene Regulatory Networks , Genetic Predisposition to Disease , Genome-Wide Association Study , Humans , Inflammation/genetics , Inflammation/metabolism , Models, Genetic , NF-kappa B/genetics , NF-kappa B/metabolism , Protein Interaction Mapping , Signal TransductionABSTRACT
BACKGROUND: Evidence suggests that chronic stimulation of beta(2)-adrenergic receptors (beta(2)-ARs) induces receptor tolerance that limits the efficacy of beta-agonists in the treatment of asthma. The precise mechanisms that induce beta(2)-AR tolerance remain unclear. OBJECTIVE: We sought to determine whether steroids modulate albuterol-induced beta(2)-AR tolerance in human small airways. METHODS: beta(2)-AR responsiveness to isoproterenol was characterized in human precision-cut lung slices (PCLSs) precontracted to carbachol after pretreatment with albuterol. RESULTS: Incubation of PCLSs with albuterol for 3, 6, or 12 hours attenuated subsequent isoproterenol-induced relaxation in a dose- and time-dependent manner. A 40% decrease (P < .0001) in maximum relaxation and a 45% decrease (P = .0011) in airway sensitivity from control values occurred after the maximum time and concentration of albuterol incubation. Desensitization was not evident when airways were relaxed to forskolin. Dexamethasone pretreatment of PCLSs (1 hour) prevented albuterol-induced beta(2)-AR desensitization by increasing the maximum drug effect (P = .0023) and decreasing the log half-maximum effective concentration values (P < .0001) from that of albuterol alone. Albuterol (12-hour incubation) decreased the beta(2)-AR cell-surface number (P = .013), which was not significantly reversed by 1 hour of preincubation with dexamethasone. CONCLUSION: These data suggest that beta(2)-AR desensitization occurs with prolonged treatment of human small airways with albuterol through mechanisms upstream of protein kinase A and that steroids prevent or reverse this desensitization. Clarifying the precise molecular mechanisms by which beta(2)-AR tolerance occurs might offer new therapeutic approaches to improve the efficacy of bronchodilators in asthma and chronic obstructive pulmonary disease.