Sputum RNA signature in allergic asthmatics following allergen bronchoprovocation test.

Autor:	Zuiker RG; Centre for Human Drug Research, Leiden, The Netherlands; RZuiker@chdr.nl., Tribouley C; Merck Research Laboratories, Rahway, New Jersey, USA.; Novartis, New York, NY, USA., Diamant Z; Centre for Human Drug Research, Leiden, The Netherlands.; Department of Respiratory Medicine and Allergology, Skane University Hospital, Lund, Sweden.; Department of Clinical & Pharmacology, University Medical Center Groningen, Groningen, The Netherlands.; Department of General Practice, University Medical Center Groningen, Groningen, The Netherlands.; QPS Netherlands, Groningen, The Netherlands., Boot JD; Centre for Human Drug Research, Leiden, The Netherlands.; Janssen Biologics B.V., Leiden, The Netherlands., Cohen AF; Centre for Human Drug Research, Leiden, The Netherlands., Van Dyck K; Merck Research Laboratories, Rahway, New Jersey, USA., De Lepeleire I; Merck Research Laboratories, Rahway, New Jersey, USA., Rivas VM; Merck Research Laboratories, Rahway, New Jersey, USA., Malkov VA; Merck Research Laboratories, Rahway, New Jersey, USA., Burggraaf J; Centre for Human Drug Research, Leiden, The Netherlands., Ruddy MK; Merck Research Laboratories, Rahway, New Jersey, USA.; EMD Serono, Rockland, MA, USA.
Jazyk:	angličtina
Zdroj:	European clinical respiratory journal [Eur Clin Respir J] 2016 Jul 13; Vol. 3, pp. 31324. Date of Electronic Publication: 2016 Jul 13 (Print Publication: 2016).
DOI:	10.3402/ecrj.v3.31324
Abstrakt:	Background: Inhaled allergen challenge is a validated disease model of allergic asthma offering useful pharmacodynamic assessment of pharmacotherapeutic effects in a limited number of subjects. Objectives: To evaluate whether an RNA signature can be identified from induced sputum following an inhaled allergen challenge, whether a RNA signature could be modulated by limited doses of inhaled fluticasone, and whether these gene expression profiles would correlate with the clinical endpoints measured in this study. Methods: Thirteen non-smoking, allergic subjects with mild-to-moderate asthma participated in a randomised, placebo-controlled, 2-period cross-over study following a single-blind placebo run-in period. Each period consisted of three consecutive days, separated by a wash-out period of at least 3 weeks. Subjects randomly received inhaled fluticasone ((FP) MDI; 500 mcg BID×5 doses in total) or placebo. On day 2, house dust mite extract was inhaled and airway response was measured by FEV1 at predefined time points until 7 h post-allergen. Sputum was induced by NaCl 4.5%, processed and analysed at 24 h pre-allergen and 7 and 24 h post-allergen. RNA was isolated from eligible sputum cell pellets (<80% squamous of 500 cells), amplified according to NuGEN technology, and profiled on Affymetrix arrays. Gene expression changes from baseline and fluticasone treatment effects were evaluated using a mixed effects ANCOVA model at 7 and at 24 h post-allergen challenge. Results: Inhaled allergen-induced statistically significant gene expression changes in sputum, which were effectively blunted by fluticasone (adjusted p<0.025). Forty-seven RNA signatures were selected from these responses for correlation analyses and further validation. This included Th2 mRNA levels for cytokines, chemokines, high-affinity IgE receptor FCER1A, histamine receptor HRH4, and enzymes and receptors in the arachidonic pathway. Individual messengers from the 47 RNA signatures correlated significantly with lung function and sputum eosinophil counts. Conclusion: Our RNA extraction and profiling protocols allowed reproducible assessments of inflammatory signatures in sputum including quantification of drug effects on this response in allergic asthmatics. This approach offers novel possibilities for the development of pharmacodynamic (PD) biomarkers in asthma.
Databáze:	MEDLINE
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