Understanding pathogenesis to develop treatments for respiratory diseases Phil Hansbro Research Centre for Healthy Lungs Univ of Newcastle & Hunter Medical Research Institute NSW, Australia 1
Aims Aim to develop effective preventions & treatments for COPD, asthma, infections, lung cancer Develop representative mouse models of human disease, interrogate, find new targets & develop new pharmaceutics Parallel human ex vivo & in vitro studies validate 2
Mouse model of COPD Directly expose airways to CS, smoke 2h/day, 5 d/wk, BALB/c or C57BL/6 mice Wk 0 8 12 Air controls Cigarette smoke treated 3 Beckett J Allergy Clin Immunol 2013, Hansbro JBC 2014, Franklin Nat Immunol 3014, Hsu AJRCCM 2015, Haw Mucosal Immunol 2016, Liu JCI Insight 2017
numbers Develops hallmark features of Human COPD * Cells/ml BALF (10 4 ) 10 8 6 ** 4 2 Air Smk 0 8 12 M N L M N L 0.0 4 d 8 wks atment TNFa (relative expression) 0.6 0.4 0.2 4wk 6wk 8wk 12wk MSCs (fold change from control) 6 4 2 0 4wk ** 6wk 8wk 12wk Control COPD 100 um 100 um Beckett J Allergy Clin Immunol 2013, Hansbro J Biol Chem 2014, Franklin Nat Immunol 2014, 4 Hsu Am J Respir Crit Care Med 2015, Haw Mucosal Immunol 2016
Emphysema & reduced lung function Control Smoke Airway RI (cmh 2 O.s/ml) 0.20 0.15 0.10 0.05 0.00 4 6 8 12 weeks Transpulmonary RI (cmh 2 O.s/ml) 0.6 0.4 0.2 0.0 cells/lung lobe 50 40 30 20 Hysteresis (cmh 2 0.%maxVpl) 10 0 150 100 Alveolar diameter (mm) 40 30 20 10 Lung mast cell numbers 0 4wk * 4 6 8 412 50 0 Weeks of treatment * * 6wk 8wk 12wk Work of breathing (abitrary units) 8 6 2 0 ** Air Smk Beckett J Allergy Clin Immunol 2013, Hansbro J Biol Chem 2014, Franklin Nat Immunol 2014, 5 Hsu Am J Respir Crit Care Med 2015, Haw/Starkey Mucosal Immunol 2016, Liu JCI Insight 2017
COPD pathogenesis Findings Mast cell proteases Trypase-b & g: Beckett JACI 2013, Hansbro JBC 2014 ASC specks/inflammasomes: Franklin Nat Immunol 2014 Amine oxidase & inhibitors: Jarnicki Br J Pharm 2016 Fibulin-1c in remodelling & emphysema also IPF & asthma: Liu JCI Insight 2016 TRAIL & inhibitors: Haw/Starkey Mucosal Immunol 2016 COPD exacerbations mirna-125a/b inhibit A20 & MAVS to promote viral infection in COPD: Hsu JCI Insight 2017 SOCS5 suppresses influenza in COPD by inhibiting EGFR: Kedzierski elife 2017 Ongoing Microbiomes: Budden Nat Rev Microbiol 2017 mirnas, acetylation, proteomics, immunometabolism, ox stress 6
Refined model DEX (d32, 33, 34) d35 Endpoints d0 Sal/Ova i.p. d12, 13 Ova i.n. Cmu/Hinf/Flu SPG/Cmu (d14) /RSV Groups Ova Ova/DEX Ova/Cmu, Ova/Hinf, Ova/Flu, Ova/RSV Ova/Cmu/DEX, Ova/Hinf/DEX, Ova/Flu/DEX, Ova/RSV/DEX d33, 34 Ova i.n. Sham-infected controls receive vehicle for infections (SPG, PBS, Media) 7 Essilfie, Horvat Thorax 2015, Kim, Horvat J Allergy Clin Immunol
Chlamydia respiratory infection induces severe, neutrophilic, steroid-resistant AAD (SSRAAD) Chlamydia-induced SSRAAD: T H 1/T H 17-associated responses neutrophils, eosinophils Steroid-resistant airway inflammation & AHR Essilfie, Horvat Thorax 2015, Kim, Horvat J Allergy Clin Immunol 2017 8
Investigation of Infection-associated, steroid-resistant asthma Chlamydia-induced neutrophilic SSRAAD Th1/Th17 & Th2 responses Haemophilus-induced neutrophilic SSRAAD Th17 & Th2 responses Influenza & RSV -induced eosinophilic SSRAAD Th1 & Th2 responses Used to discover mechanisms of pathogenesis & treatment targets (mirnas, epigenetics, inflammasomes) Microarrays & RNA-seq to correlate with human phenoptypes 9
Severe asthma Essilfie Thorax 2015 Kim, Horvat J Allergy Clin Immunol 2017 Kim Am J Respir Crit Care Med 2017 10
Early life Chlamydia respiratory infection-induced chronic lung disease in mice 0 day, 3 week, 6 week C. muridarum profile 9wk old C. muridarum (Cmu) or sham-inoculation (SPG) Infection only Day 0 20 Inflammation profile 45 61 Assess lung structure and function Ova sensitisation Ova challenge Ova-induced AAD 45 57-60 CS-exposure 16wk old CS-induced experimental COPD Horvat AJRCCM 2007 Horvat/Starkey JACI 2010 Starkey/Haw Mucosal Immunol 2015 Starkey AJRCMB 2016 45 112
Published & ongoing studies Neonatal & infant infection infection causes long term changes: immunity (switches T cell phenotype, modifies DC function), alveolar enlargement AHR increases severity of subsequent AAD In infants roles for IL-13 & TRAIL, inhibition suppresses features & subsequent AAD Horvat AJRCCM 2007, Horvat/Starkey JACI 2010, Starkey Mucosal Immunol 2013, 2014 IL-13, ILCs, TLR2, epigenetics, proteomics 12
Summary Early-life infection TLR2 IL-13 + ILC2s IL-13Rα1 Stat6 mir-21 Impaired lung function
Early-life asthma Jay Horvat Malcolm Starkey Richard Kim Duc Nguyen Severe asthma Jay Horvat Richard Kim James Pinkerton Ama Essilfie COPD Mike Fricker Emma Beckett Andrew Jarnicki Kurtis Budden Kamal Dua Rest of Hansbro group NHMRC, Asthma Fnd NSW, ARC Newcastle Paul Foster Peter Gibson & Katie Baines Peter Wark Darryl Knight Joerg Mattes Mast cells Rick Stevens - Harvard Microbiomes Phil Hugenholtz UQ Paul Dennis - UQ Mark Morrison UQ Inflammasomes Matt Cooper UQ Avril Robertson - UQ Luke O Neill Trinity Coll Ireland Eicke Latz Univ Bonn Omics data Sally Wenzel - Pittsburgh Ian Adcock Newcastle/Imperial DFCO Wayne Metzner Johns Hopkins 14