Non-neuronal cholinergic system contributes to corticosteroid resistance in chronic obstructive pulmonary disease patients

Abstract

BackgroundInhaled corticosteroid (ICS) with long-acting beta-2 agonists is a well-documented combination therapy for chronic obstructive pulmonary disease (COPD) based on its additive anti-inflammatory properties. By contrast, the recommendation of ICS in combination with long-acting muscarinic antagonist (LAMA) is not evidence-based. In this study, neutrophils obtained from COPD patients were used to compare the anti-inflammatory effects of aclidinium bromide (a long-acting muscarinic antagonist) with corticosteroids and their potential additive effect.MethodsHuman sputum and blood neutrophils were isolated from healthy individuals (n = 37), patients with stable COPD (n = 52) and those with exacerbated COPD (n = 16). The cells were incubated with corticosteroid fluticasone propionate (0.1 nM–1 μM), aclidinium bromide (0.1 nM–1 μM) or a combination thereof and stimulated with 1 μg of lipopolysaccharide/ml or 5 % cigarette smoke extract. Levels of the pro-inflammatory mediators interleukin-8, matrix metalloproteinase-9, CCL-5, granulocyte-macrophage colony-stimulating factor and interleukin-1β were measured and the mechanisms of corticosteroid resistance evaluated at the end of the incubation.ResultsThe non-neuronal cholinergic system was over-expressed in neutrophils from COPD patients, as evidenced by increases in the expression of muscarinic receptors (M2, M4 and M5), choline acetyltransferase and vesicular acetylcholine transporter. Aclidinium bromide demonstrated anti-inflammatory effects on neutrophils from COPD patients, reversing their resistance to corticosteroids. Additive effects of combined aclidinium bromide and fluticasone propionate in blocking M2 receptor levels, inhibiting phosphoinositide 3-kinase-δ and enhancing the glucocorticoid response element transcription factor were demonstrated and were accompanied by an increase in the corticosteroid-induced expression of anti-inflammatory-related genes.ConclusionsLAMAs potentiate the anti-inflammatory effects of corticosteroids in neutrophils from COPD patients in vitro, thus providing a scientific rationale for their use in combination with corticosteroids in the treatment of COPD.Electronic supplementary materialThe online version of this article (doi:10.1186/s12931-016-0467-8) contains supplementary material, which is available to authorized users.

In vitro anti-inflammatory effects of AZD8999, a novel bifunctional muscarinic acetylcholine receptor antagonist /β2-adrenoceptor agonist (MABA) compound in neutrophils from COPD patients

“…Peripheral blood neutrophils were isolated from peripheral venous blood and cultured as previously described [9]. Neutrophils were adjusted to 500×10 3 cells per well in 24-well plates and incubated in RPMI 1640 for 1 h at 37°C, 5% CO 2 .…”

“…Although evidence for drug combinations comes from clinical trials, scientific rationale for drug combinations can be explained by molecular interactions as previously outlined [5–8]. Recent evidence provided by our group indicates that LAMA can improve corticosteroid insensitivity in neutrophils from COPD patients inhibiting PI3Kδ and enhancing glucocorticoid response element transcription factor (GRE) with the consequent increased expression of corticosteroid anti-inflammatory related genes [9]. Although not studied in detail, these results indicate potential anti-inflammatory synergism between triple therapy based in LABA + LAMA + ICS.…”

Is aclidinium alone or combined with a LABA a rational choice for symptomatic COPD patients?

“…For example, there is some evidence that aclidinium bromide could modulate the non-neuronal cholinergic system, a recently identified regulatory pathway in pulmonary inflammation and remodeling [28]. Dysfunction of the non-neuronal cholinergic system seems to be involved in the pathophysiology of COPD and to some extent in resistance to corticosteroid therapy; in fact, non-neuronal cholinergic system is over-expressed in corticosteroid-insensitive neutrophils from COPD patients, as evidenced by increases in the expression of muscarinic receptors (M2, M4 and M5), vesicular acetylcholine transporter (VAChT) and choline acetyltransferase (ChAT).…”

“…Mitogen-activated Protein Kinase Phosphatase 1 (MKP1), Cysteine-RIch Secretory Protein LCCL Domain-containing 2 (CRISPLD2) and (GILZ) [28]. Among aclidinium bromide therapeutic properties, it seems to be interesting as well its favourable effect on hyperinflation and airflow limitation, that plays a very important pathophysiological role in inducing dyspnea and reducing quality of life (QoL) in COPD patients; in this regard, it has been recently demonstrated that aclidinium promotes a rapid and relevant desufflation and improves lung ventilation inhomogeneity, even in severe/very severe COPD patients [29].…”

Effects of Glutathione S‐Transferase Gene Polymorphisms and Antioxidant Capacity per Unit Albumin on the Pathogenesis of Chronic Obstructive Pulmonary Disease

“…Airway remodeling is associated with the process of airway inflammation, which predominately occurs in the lungs [ 3 ]. Since the pathogenesis of COPD is not yet fully understood [ 4 ], such research is typically focused on four aspects which include (1) lung inflammation, (2) the imbalance of lung protease and anti-proteases, (3) oxidation and antioxidation imbalances, and (4) autonomic nervous system dysfunction [ 5 ], such as abnormal cholinergic receptor distribution ( Figure 1 ).…”