Rhiannon B Werder1,2,3,4, Md Ashik Ullah1, Muhammed Mahfuzur Rahman1,2, Jennifer Simpson1,2,5, Jason P Lynch1,6,7, Natasha Collinson1, Sonja Rittchen1,8, Ridwan B Rashid1,2, Md Al Amin Sikder1,2, Herlina Y Handoko1, Bodie F Curren1,2, Ismail Sebina1, Gunter Hartel1, Alec Bissell1, Sylvia Ngo1, Tejasri Yarlagadda9, Sumaira Z Hasnain10,11, Wenying Lu12, Sukhwinder S Sohal12, Megan Martin13, Simon Bowler11,13, Lucy D Burr11,13, Laurent O Martinez14, Bernard Robaye15, Kirsten Spann9, Manuel A R Ferreira1, Simon Phipps1,2,10. 1. Respiratory Immunology Laboratory, QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia. 2. Faculty of Medicine and. 3. Center for Regenerative Medicine, Boston University and Boston Medical Center, Boston, Massachusetts. 4. The Pulmonary Center, Department of Medicine, School of Medicine, Boston University, Boston, Massachusetts. 5. Barrier Immunity Section, Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, Maryland. 6. Center for Bacterial Pathogenesis, Division of Infectious Diseases, Department of Medicine, Massachusetts General Hospital, Boston, Massachusetts. 7. Department of Microbiology, Blavatnik Institute, Harvard Medical School, Harvard University, Boston, Massachusetts. 8. Otto Loewi Research Center for Vascular Biology, Immunology and Inflammation, Division of Pharmacology, Medical University of Graz, Graz, Austria. 9. School of Biomedical Sciences, Queensland University of Technology, Brisbane, Queensland, Australia. 10. Australian Infectious Diseases Research Centre, University of Queensland, Brisbane, Queensland, Australia. 11. Immunopathology Group, Mater Research Institute-University of Queensland, Translational Research Institute, Brisbane, Queensland, Australia. 12. Respiratory Translational Research Group, Department of Laboratory Medicine, School of Health Sciences, College of Health and Medicine, University of Tasmania, Launceston, Tasmania, Australia. 13. Department of Respiratory Medicine, Mater Adult Hospital, South Brisbane, Queensland, Australia. 14. Institut des Maladies Métaboliques et Cardiovasculaires, Université de Toulouse, Institut National de la Santé et de la Recherche Médicale, and Université Toulouse III-Paul Sabatier, Toulouse, France; and. 15. Institute of Interdisciplinary Research, Free University of Brussels, Gosselies, Belgium.
Abstract
Rationale: The alarmins IL-33 and HMGB1 (high mobility group box 1) contribute to type 2 inflammation and asthma pathogenesis. Objectives: To determine whether P2Y13-R (P2Y13 receptor), a purinergic GPCR (G protein-coupled receptor) and risk allele for asthma, regulates the release of IL-33 and HMGB1. Methods: Bronchial biopsy specimens were obtained from healthy subjects and subjects with asthma. Primary human airway epithelial cells (AECs), primary mouse AECs, or C57Bl/6 mice were inoculated with various aeroallergens or respiratory viruses, and the nuclear-to-cytoplasmic translocation and release of alarmins was measured by using immunohistochemistry and an ELISA. The role of P2Y13-R in AEC function and in the onset, progression, and exacerbation of experimental asthma was assessed by using pharmacological antagonists and mice with P2Y13-R gene deletion. Measurements and Main Results: Aeroallergen exposure induced the extracellular release of ADP and ATP, nucleotides that activate P2Y13-R. ATP, ADP, and aeroallergen (house dust mite, cockroach, or Alternaria antigen) or virus exposure induced the nuclear-to-cytoplasmic translocation and subsequent release of IL-33 and HMGB1, and this response was ablated by genetic deletion or pharmacological antagonism of P2Y13. In mice, prophylactic or therapeutic P2Y13-R blockade attenuated asthma onset and, critically, ablated the severity of a rhinovirus-associated exacerbation in a high-fidelity experimental model of chronic asthma. Moreover, P2Y13-R antagonism derepressed antiviral immunity, increasing IFN-λ production and decreasing viral copies in the lung. Conclusions: We identify P2Y13-R as a novel gatekeeper of the nuclear alarmins IL-33 and HMGB1 and demonstrate that the targeting of this GPCR via genetic deletion or treatment with a small-molecule antagonist protects against the onset and exacerbations of experimental asthma.
Rationale: The alarmins IL-33 and HMGB1 (high mobility group box 1) contribute to type 2 inflammation and asthma pathogenesis. Objectives: To determine whether P2Y13-R (P2Y13 receptor), a purinergic GPCR (G protein-coupled receptor) and risk allele for asthma, regulates the release of IL-33 and HMGB1. Methods: Bronchial biopsy specimens were obtained from healthy subjects and subjects with asthma. Primary human airway epithelial cells (AECs), primary mouse AECs, or C57Bl/6 mice were inoculated with various aeroallergens or respiratory viruses, and the nuclear-to-cytoplasmic translocation and release of alarmins was measured by using immunohistochemistry and an ELISA. The role of P2Y13-R in AEC function and in the onset, progression, and exacerbation of experimental asthma was assessed by using pharmacological antagonists and mice with P2Y13-R gene deletion. Measurements and Main Results: Aeroallergen exposure induced the extracellular release of ADP and ATP, nucleotides that activate P2Y13-R. ATP, ADP, and aeroallergen (house dust mite, cockroach, or Alternaria antigen) or virus exposure induced the nuclear-to-cytoplasmic translocation and subsequent release of IL-33 and HMGB1, and this response was ablated by genetic deletion or pharmacological antagonism of P2Y13. In mice, prophylactic or therapeutic P2Y13-R blockade attenuated asthma onset and, critically, ablated the severity of a rhinovirus-associated exacerbation in a high-fidelity experimental model of chronic asthma. Moreover, P2Y13-R antagonism derepressed antiviral immunity, increasing IFN-λ production and decreasing viral copies in the lung. Conclusions: We identify P2Y13-R as a novel gatekeeper of the nuclear alarmins IL-33 and HMGB1 and demonstrate that the targeting of this GPCR via genetic deletion or treatment with a small-molecule antagonist protects against the onset and exacerbations of experimental asthma.
Entities:
Keywords:
GPCR; alarmin; epithelium; pneumonia virus of mice; purinergic; rhinovirus