How can human models of ALI inform clinical trials
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1 How can human models of ALI inform clinical trials Danny McAuley Royal Victoria Hospital and Queen s University of Belfast Critical Care Cananda Forum October 2012
2 Disclosures GSK; consultancy and participate in research funded by GSK
3 How can human models of ALI inform clinical trials Limitations of animal models Human models of ALI Ex-vivo lung perfusion LPS inhalation Surgical models of ALI Identifying novel therapies using human models
4 McAuley et al. CCM 38(10):S523 Stepwise approach
5 NEJM : Translating animal data to man
6 The simple truths Human cells are not Animal cells Cancer or virally transformed cell lines Humans are not Animals
7 Monocyte BAL neutrophils depletion reduces & total protein pulmonary inflammation in a murine ALI model Dhaliwal et al. AJRCCM (in press)
8 Monocyte BAL neutrophils depletion & does total NOT protein reduce pulmonary inflammation in a human ALI model MD arm n=15 Sham arm n=14
9 New models of ALI Surgical models LPS model Human Ex-Vivo Lung Perfusion
10 Human ex-vivo lung perfusion
11 Human ex-vivo lung perfusion CPAP 10 cm H O 95% O, 5% CO Peristaltic pump Lung Pulmonary Artery Pressure = mm Hg Perfusate Reservoir 37 C water bath Left Atrial Pressure = 0 mm Hg Lee JW et al. PNAS :
12 Human ex-vivo lung perfusion Surgical preparation of lung Begin perfusion without blood Intervention AFC and BAL in RUL or LUL (Control) AFC and BAL in RML or LLL (Endotoxin) 1 hr 1 hr 4 hrs - Lung temp 36 C - Apply CPAP Add 100 ml fresh whole blood to perfusate Instill 0.1 mg/kg of endotoxin into the airspaces of the RML or LLL
13 Histological evidence of ALI Control Lung Lobe LPS Lung Lobe Absolute Neutrophil Counts 6 9 ± 6 x 10 cells 6 25 ± 25 x 10 cells *
14 Impaired alveolar fluid clearance Alveolar fluid clearance (%/h) * 0 Control No Blood Blood + Endotoxin
15 EVLWa Increased pulmonary oedema is associated with mortality in ALI 35 EVLW (ml/kg) * 0 non survivor survivor Craig et al. CCM 2011
16 IL-1 (pg/ml) TNF (pg/ml) IL-8 (pg/ml) Cytokine response in ex-vivo lung consistent with ALI * * * Control LPS (0.1 mg/kg) 0 Control LPS (0.1 mg/kg) 0 Control LPS (0.1 mg/kg)
17 Mesenchymal stem cells (MSC) improves pulmonary inflammation Control Lung Lobe Absolute Neutrophil Counts LPS Lung Lobe LPS + MSC Lung Lobe LPS + MSC conditioned media Lung Lobe 6 9 ± 6 x 10 cells 6 25 ± 25 x 10 cells 6 13 ± 11 x 10 cells 6 6 ± 5 x 10 cells * P = P =
18 Human ex-vivo lung perfusion In vivo lung injury Surgical preparation of lung Perfusion Baseline AFC (Control) AFC (Treatment) 1 hr 4 hrs - Lung temp 36 C - Apply CPAP Intervention
19 Alveolar Fluid Clearance (%/h) MSCs improve alveolar fluid clearance and is mediated by KGF P< * 30 * Control (AFC at 0 hr) + IV MSC AFC at 4 hr + Control Goat IgG + IV MSC AFC at 4 hr + Goat Anti-KGF Ab * P significant by ANOVA vs. Control P significant by ANOVA vs. MSC IV
20 Inhaled LPS as an in vivo model to study pulmonary inflammation in healthy subjects FEV1 BAL FEV1 Plasma 6 hr 18 hr FEV1 Plasma 50 µg LPS inhalation E. Coli serotype O26:B6 (Sigma) Breath activated dosimeter
21 Cecilia O Kane Inhaled LPS induces inflammatory cytokines in pulmonary compartment
22 Cecilia O Kane Inhaled LPS induces pulmonary epithelial activation/injury
23 Cecilia O Kane Increased BAL protein as a measure of alveolar barrier function
24 Simvastatin in the inhaled LPS model of ALI Treatment with a clinically relevant dose of simvastatin will reduce pulmonary inflammation induced by LPS inhalation in humans Shyamsundar et al. AJRCCM :
25 Schedule for patient safety monitoring
26 Simvastatin decreases pulmonary TNF following LPS inhalation * p < 0.05 vs placebo
27 Lovastatin decreases pulmonary inflammation measured by FDG PET following LPS instillation Chen et al. AJRCCM :
28 Elective surgery as a cause of acute lung injury ALI/ARDS associated with: Oesophagectomy 15-40% Lung resection 1-7% Cardiopulmonary bypass (CPB) 1.3% Elective AAA repair
29 One lung ventilation as a model of direct pulmonary injury Overdistension Hyperperfusion Hyperoxia Ischaemiareperfusion Atelectotrauma Surgical trauma (contusion & lymphatic disruption) Local and systemic release of inflammatory mediators
30 Pulmonary inflammation associated with one lung ventilation Cooling tube and insulating jacket Connection to patient via endo-tracheal tube Expiratory flow to ventilator RTube, Respiratory Research Inc One-way valve Inspiratory flow from ventilator Moloney et al AJRCCM 169:64
31 Simvastatin decreases alveolar epithelial and systemic endothelial injury during one lung ventilation
32 Cardiopulmonary bypass as a systemic model causing pulmonary inflammation Moloney et al AJRCCM 169:64
33 AAA repair induces systemic and pulmonary inflammation
34 AAA repair associated with impaired oxygenation
35 Limitations of models Model specific mechanisms LPS induced inflammation Ischaemia reperfusion Short-term Need to use to test treatment as well as pretreatment interventions
36 Conclusions Variety of human models characterised by pulmonary and systemic injury reflecting ALI Useful in testing novel therapeutic agents to identify ineffective therapy Improve design of subsequent clinical trials of pharmacological agents
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