Interstitial lung disease : from bench to bedside

printing supported by . Visit Chiesi at Stand B2.10 WEDNESDAY, SEPTEMBER 5TH 2012 4530 LSC 2012 Abstract – The cell-penetrating P1pal-12 pepducin limits pulmonary fibrosis in the murine bleomycin model C. Lin, J.W. Duitman, K. Borensztajn, C.A. Spek. CEMM, Academisch Medisch Centrum (AMC), Amsterdam, Netherlands Idiopathic pulmonary fibrosis is the most devastating fibrotic diffuse parenchymal lung disease which remains refractory to available pharmacological therapies.Therefore, novel treatment options are urgently required. Protease-activated receptor (PAR)-1 is a heptahelical G protein-coupled receptor that mediates critical signaling pathways in pathology. Interestingly, bleomycin-induced lung fibrosis was shown to be diminished in PAR-1 deficient mice. We thus hypothesized that pharmacological PAR-1 inhibition may be an interesting therapeutic approach to combat pulmonary fibrosis. Consequently, we explored the effect of P1pal-12 (a pepducin blocking the PAR-1/G-protein interaction) during the development of lung fibrosis induced by intranasal instillation of bleomycin. We show that once daily treatment with 0.5, 2.5 or 10 mg/kg P1pal-12, reduced severity and extent of fibrotic lesions in a dose-dependent manner (2.5 and 2 fold reduction with 2.5 and 10 mg/kg). These findings correlated with significant decreases in fibronectin, collagen and α-SMA mRNA expression levels in treated mice. Moreover, fibrin deposition in the lungs was reduced by 26% ± 3% (p<0.05) in 2.5 mg/kg treated mice compared to untreated controls. Finally,P1pal-12 reduced bleomycin-induced IL-6 and MCP-1 levels in lung homogenates by 65±3% (p<0.01) and 36±3% (p<0.05) respectively. Overall, our data show that P1pal-12 limits lung fibrosis suggesting that targeting PAR-1 may be a promising therapeutic strategy for pulmonary fibrosis. 4531 Effect of nintedanib on silica-induced lung inflammation and fibrosis in mice Isabelle Maillet1, Valérie Quesniaux1, Lutz Wollin2, Bernhard Ryffel1. 1Molecular Immunology, INEM UMR7355, CNRS, Orleans, France; 2Respiratory Diseases Research, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany Introduction: One-year treatment with the receptor tyrosine kinase inhibitor nintedanib (BIBF 1120) specific for vascular endothelial growth factor receptor (VEGFR), platelet-derived growth factor receptor (PDGFR) and fibroblast growth factor receptor (FGFR) was associated with a 68.4% reduction in the rate of decline of forced vital capacity in patients with idiopathic pulmonary fibrosis (IPF) versus placebo, which approached statistical significance. Aim: To further explore its mode of action, nintedanib was tested in a mouse model of silicosis displaying ongoing pulmonary inflammation and fibrosis. Methods: Within 30 days a single intranasal administration of silica caused a robust lung inflammation with a significant increase in macrophages, neutrophils and lymphocytes in the BALF, increased IL-1 beta, CXCL1/KC and TIMP1 production, and increased collagen deposition in the lung. Histologic analysis revealed chronic inflammation with granuloma formation and fibrosis demonstrated by collagen staining. Results: Nintedanib administered by gavage at 30 and 100 mg/kg/day significantly reduced neutrophil and lymphocyte counts, but had no effect on macrophage counts in the BALF. Furthermore, IL-1 beta, CXCL1/KC, TIMP1, collagen in lung and lung inflammation with granuloma and fibrosis were drastically reduced. Conclusion: Nintedanib effectively reduced silica-induced chronic inflammation and fibrosis in mice. The anti-inflammatory and anti-fibrotic features of nintedanib may impact the progressive course of fibrotic lung diseases like IPF or silicosis. 4532 Telomere (TL) shortening is associated with disease severity in scleroderma (SSC) associated interstitial lung disease (ILD) Huzaifa I. Adamali, Cesar M. Delgado, Carmel Stock, Gisela E. Lindhal, Phil Molyneaux, Anne-Marie Russell, Athol Wells, Elizabeth A. Renzoni, Toby Maher. Interstitial Lung Disease, Royal Brompton Hospital and National Heart Lung Institute, Imperial College, London, United Kingdom Attrition of TL is associated with the development and progression of pulmonary fibrosis. TL length has been shown to be reduced in individuals with SSC vs healthy controls. ILD develops in over 25% of individuals with SSC. We hypothesized that TL shortening is an important mechanism driving the pathogenesis of SSCILD. Methods: Whole blood was collected from SSC-ILD patients (n=132). SSC-ILD was defined as extensive or limited disease (Goh et al, AJRCCM 2008;177). DNA, isolated using a Promega extraction kit, was analysed using quantitative real time PCR. TL length was calculated using the method described by O’Callaghan and Fenech (Biol Proced. 2011; 31). Results: Mean±SEM TL length in SSC cohort was 65.1±4.7 kb/diploid genome. In limited disease (n=100, 74 female, age 53.2±1.1 yrs) mean TL length was 77.0±5.5 kb/diploid genome (see figure 1). In extensive disease (n=32, 18 females, age 46.5±3.5 yrs), the mean TL length was 24.8±3.5 kb/diploid genome(p<0.0001). TL length correlated with extent of fibrosis on CT (p<0.001). TL typically shorten with age. In our cohort TL length increased with age(p=0.02) perhaps reflecting a trend towards more extensive disease in younger subjects. Conclusion: TL length is significantly associated with disease extent in SSC-ILD. Figure 1. Comparison of telomere length in individuals with limited and extensive SSC associated ILD. Box and whisker plot demonstrating mean and 95% confidence intervals with outliers. Our observation suggests an important role for premature cellular senescence in the pathogenesis of SSC-ILD. HA is an ERS-Fellow. 4533 HSP47siRNA targeting to myofibroblasts attenuates bleomycin-induced pulmonary fibrosis Mitsuo Otsuka, Masanori Shiratori, Koji Kuronuma, Hiroki Takahashi. Third Department of Internal Medicine, Sapporo Medical University School of Medicine, Sapporo, Japan Rationale: The 47-kDa heat shock protein 47 (HSP47) plays a role in the processing of procollagens as a collagen molecular chaperone. The HSP47-positive cells increase in lung fibrosis, which suggests an important role of HSP47 in the pathogenesis of lung fibrosis. It is expected that to control the expression of the HSP47 may reduce the lung fibrosis. Lung myofibroblasts are speculated vitamin A (VA)-storing cells in the lungs like hepatic stellate cells. Based on these properties of myofibroblasts, we prepared a VA-coupled liposomes carrying HSP47siRNA. The goal of this study is to investigate whether the inhibition of HSP47 for the myofibroblasts with siRNA influences to lung fibrosis. Methods: Sprague-Dawley male rats were treated with intra-tracheal bleomycin (BLM) or PBS. We injected VA-coupled liposomes carrying HSP47siRNA into rat tail vein three times in a week. We made 5 groups; PBS intratracheally & PBS intravenously, BLM & PBS, BLM & VA-HSP47siRNA, BLM & HSP47siRNA, and BLM & VA-randomRNA, respectively. We measured the contents of lung hydroxyproline, BAL cells counts, HSP47 expression, and cytokines. Results: Western blotting showed the expressions of HSP47 were reduced in rats treated with VA-coupled liposomes carrying HSP47siRNA. The liposomes with HSP47siRNA significantly improved lung fibrosis morphologically and the increases of hydroxyproline contents in the lungs, the inflammatory cytokines, and the number of BAL cells. Conclusions: These results suggest that HSP47siRNA improves bleomycininduced lung fibrosis and this drug delivery system is useful methods. 826s Oral Presentation Room A3 08:30 10:30 Abstract printing supported by . Visit Chiesi at Stand B2.10printing supported by . Visit Chiesi at Stand B2.10