Christos Rossios1, Stelios Pavlidis2, Uruj Hoda1, Chih-Hsi Kuo3, Coen Wiegman1, Kirsty Russell1, Kai Sun3, Matthew J Loza4, Frederic Baribaud4, Andrew L Durham1, Oluwaseun Ojo1, Rene Lutter5, Anthony Rowe4, Aruna Bansal6, Charles Auffray7, Ana Sousa8, Julie Corfield9, Ratko Djukanovic10, Yike Guo3, Peter J Sterk5, Kian Fan Chung1, Ian M Adcock11. 1. Airways Disease, National Heart & Lung Institute, Imperial College London, and the Biomedical Research Unit, Royal Brompton & Harefield NHS Trust, London, United Kingdom. 2. Department of Computing & Data Science Institute, Imperial College London, London, United Kingdom; Janssen Research and Development, High Wycombe, United Kingdom. 3. Department of Computing & Data Science Institute, Imperial College London, London, United Kingdom. 4. Janssen Research and Development, High Wycombe, United Kingdom. 5. Faculty of Medicine, University of Amsterdam, Amsterdam, The Netherlands. 6. Acclarogen, St John's Innovation Centre, Cambridge, United Kingdom. 7. European Institute for Systems Biology and Medicine, CNRS-ENS-UCBL, Université de Lyon, Lyon, France. 8. Respiratory Therapeutic Unit, GSK, Stockley Park, United Kingdom. 9. AstraZeneca R&D, Molndal, Sweden, and Areteva R&D, Nottingham, United Kingdom. 10. Faculty of Medicine, Southampton University, and the NIHR Southampton Respiratory Biomedical Research Unit, University Hospital Southampton, Southampton, United Kingdom. 11. Airways Disease, National Heart & Lung Institute, Imperial College London, and the Biomedical Research Unit, Royal Brompton & Harefield NHS Trust, London, United Kingdom; Priority Research Centre for Healthy Lungs, Hunter Medical Research Institute, University of Newcastle, Newcastle, Australia. Electronic address: ian.adcock@imperial.ac.uk.
Abstract
BACKGROUND: Sputum analysis in asthmatic patients is used to define airway inflammatory processes and might guide therapy. OBJECTIVE: We sought to determine differential gene and protein expression in sputum samples from patients with severe asthma (SA) compared with nonsmoking patients with mild/moderate asthma. METHODS: Induced sputum was obtained from nonsmoking patients with SA, smokers/ex-smokers with severe asthma, nonsmoking patients with mild/moderate asthma (MMAs), and healthy nonsmoking control subjects. Differential cell counts, microarray analysis of cell pellets, and SOMAscan analysis of sputum analytes were performed. CRID3 was used to inhibit the inflammasome in a mouse model of SA. RESULTS: Eosinophilic and mixed neutrophilic/eosinophilic inflammation were more prevalent in patients with SA compared with MMAs. Forty-two genes probes were upregulated (>2-fold) in nonsmoking patients with severe asthma compared with MMAs, including IL-1 receptor (IL-1R) family and nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain containing 3 (NRLP3) inflammasome members (false discovery rate < 0.05). The inflammasome proteins nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing 1 (NLRP1), NLRP3, and nucleotide-binding oligomerization domain (NOD)-like receptor C4 (NLRC4) were associated with neutrophilic asthma and with sputum IL-1β protein levels, whereas eosinophilic asthma was associated with an IL-13-induced TH2 signature and IL-1 receptor-like 1 (IL1RL1) mRNA expression. These differences were sputum specific because no activation of NLRP3 or enrichment of IL-1R family genes in bronchial brushings or biopsy specimens in patients with SA was observed. Expression of NLRP3 and of the IL-1R family genes was validated in the Airway Disease Endotyping for Personalized Therapeutics cohort. Inflammasome inhibition using CRID3 prevented airway hyperresponsiveness and airway inflammation (both neutrophilia and eosinophilia) in a mouse model of severe allergic asthma. CONCLUSION: IL1RL1 gene expression is associated with eosinophilic SA, whereas NLRP3 inflammasome expression is highest in patients with neutrophilic SA. TH2-driven eosinophilic inflammation and neutrophil-associated inflammasome activation might represent interacting pathways in patients with SA.
BACKGROUND: Sputum analysis in asthmatic patients is used to define airway inflammatory processes and might guide therapy. OBJECTIVE: We sought to determine differential gene and protein expression in sputum samples from patients with severe asthma (SA) compared with nonsmoking patients with mild/moderate asthma. METHODS: Induced sputum was obtained from nonsmoking patients with SA, smokers/ex-smokers with severe asthma, nonsmoking patients with mild/moderate asthma (MMAs), and healthy nonsmoking control subjects. Differential cell counts, microarray analysis of cell pellets, and SOMAscan analysis of sputum analytes were performed. CRID3 was used to inhibit the inflammasome in a mouse model of SA. RESULTS: Eosinophilic and mixed neutrophilic/eosinophilic inflammation were more prevalent in patients with SA compared with MMAs. Forty-two genes probes were upregulated (>2-fold) in nonsmoking patients with severe asthma compared with MMAs, including IL-1 receptor (IL-1R) family and nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain containing 3 (NRLP3) inflammasome members (false discovery rate < 0.05). The inflammasome proteins nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing 1 (NLRP1), NLRP3, and nucleotide-binding oligomerization domain (NOD)-like receptor C4 (NLRC4) were associated with neutrophilic asthma and with sputum IL-1β protein levels, whereas eosinophilic asthma was associated with an IL-13-induced TH2 signature and IL-1 receptor-like 1 (IL1RL1) mRNA expression. These differences were sputum specific because no activation of NLRP3 or enrichment of IL-1R family genes in bronchial brushings or biopsy specimens in patients with SA was observed. Expression of NLRP3 and of the IL-1R family genes was validated in the Airway Disease Endotyping for Personalized Therapeutics cohort. Inflammasome inhibition using CRID3 prevented airway hyperresponsiveness and airway inflammation (both neutrophilia and eosinophilia) in a mouse model of severe allergic asthma. CONCLUSION: IL1RL1 gene expression is associated with eosinophilic SA, whereas NLRP3 inflammasome expression is highest in patients with neutrophilic SA. TH2-driven eosinophilic inflammation and neutrophil-associated inflammasome activation might represent interacting pathways in patients with SA.
Authors: Matthew S Godwin; Kristen M Reeder; Jaleesa M Garth; Jonathan P Blackburn; MaryJane Jones; Zhihong Yu; Sadis Matalon; Annette T Hastie; Deborah A Meyers; Chad Steele Journal: JCI Insight Date: 2019-11-01
Authors: Marrah E Lachowicz-Scroggins; Eleanor M Dunican; Annabelle R Charbit; Wilfred Raymond; Mark R Looney; Michael C Peters; Erin D Gordon; Prescott G Woodruff; Emma Lefrançais; Brenda R Phillips; David T Mauger; Suzy A Comhair; Serpil C Erzurum; Mats W Johansson; Nizar N Jarjour; Andrea M Coverstone; Mario Castro; Annette T Hastie; Eugene R Bleecker; Merritt L Fajt; Sally E Wenzel; Elliot Israel; Bruce D Levy; John V Fahy Journal: Am J Respir Crit Care Med Date: 2019-05-01 Impact factor: 21.405
Authors: A T Hastie; C Steele; C W Dunaway; W C Moore; B M Rector; E Ampleford; H Li; L C Denlinger; N Jarjour; D A Meyers; E R Bleecker Journal: Clin Exp Allergy Date: 2018-04-15 Impact factor: 5.018
Authors: Michael C Peters; Sheena Kerr; Eleanor M Dunican; Prescott G Woodruff; Merritt L Fajt; Bruce D Levy; Elliot Israel; Brenda R Phillips; David T Mauger; Suzy A Comhair; Serpil C Erzurum; Mats W Johansson; Nizar N Jarjour; Andrea M Coverstone; Mario Castro; Annette T Hastie; Eugene R Bleecker; Sally E Wenzel; John V Fahy Journal: J Allergy Clin Immunol Date: 2018-03-07 Impact factor: 10.793