Shashi Chillappagari1, Christian Müller2, Poornima Mahavadi3, Andreas Guenther4, Lutz Nährlich5, Jonathan Rosenblum6, Bruce K Rubin7, Markus O Henke8. 1. Department of Pediatrics, Justus-Liebig-University Giessen, Feulgenstrasse 12, Giessen 35392, Germany; Department of Internal Medicine, Justus-Liebig-University Giessen, Klinikstrasse-36, Giessen 35392, Germany; Department of Pulmonary Medicine, Philipps-University Marburg, Baldingerstrasse 1, Marburg 35043, Germany; German Centre for Lung Research (DZL), Giessen, Germany. 2. Department of Pulmonary Medicine, Philipps-University Marburg, Baldingerstrasse 1, Marburg 35043, Germany. 3. Department of Internal Medicine, Justus-Liebig-University Giessen, Klinikstrasse-36, Giessen 35392, Germany; German Centre for Lung Research (DZL), Giessen, Germany. 4. Department of Internal Medicine, Justus-Liebig-University Giessen, Klinikstrasse-36, Giessen 35392, Germany; German Centre for Lung Research (DZL), Giessen, Germany; European IPF Registry/Biobank, Lung Clinic Waldhof-Elgershausen, Greifenstein, Germany. 5. Department of Pediatrics, Justus-Liebig-University Giessen, Feulgenstrasse 12, Giessen 35392, Germany; German Centre for Lung Research (DZL), Giessen, Germany. 6. ActivX Biosciences, Inc., 11025 North Torrey Pines Road, Suite 120, La Jolla, CA 92037, USA. 7. Department of Pediatrics, Virginia Commonwealth University School of Medicine, 1001 East Marshall Street, Richmond, VA 23298, USA. 8. Department of Pulmonary Medicine, Philipps-University Marburg, Baldingerstrasse 1, Marburg 35043, Germany; Asklepios Fachkliniken München-Gauting, Gauting, Germany; Comprehensive Pneumology Center (CPC), Helmholtz Zentrum, Munich, Germany. Electronic address: m.henke@asklepios.com.
Abstract
BACKGROUND: Neutrophil elastase (NE) rapidly degrades gel-forming airway mucins in cystic fibrosis (CF) sputum. We hypothesized that KRP-109, a small molecule NE inhibitor, would inhibit CF mucin degradation in vitro. METHODS: Sputa were collected from CF patients (n=5) chronically or intermittently infected with Pseudomonas aeruginosa (P.a.). Mucin degradation was analyzed using western blot. Protease inhibitor studies were performed using alpha1-proteinase inhibitor (A1-PI Prolastin®) and KRP-109. Elastase activity assays were performed using spectrophotometry. RESULTS: There were significant differences in the amount of active NE in different CF sputum samples. KRP-109 decreased the NE driven mucin degradation in vitro. Pseudomonas elastases appeared to blunt elastase inhibition by A1-PI or KRP-109. CONCLUSION: Inhibitors of neutrophil and Pseudomonas-derived elastases might rescue mucus clearance and reverse airway obstruction in CF.
BACKGROUND: Neutrophil elastase (NE) rapidly degrades gel-forming airway mucins in cystic fibrosis (CF) sputum. We hypothesized that KRP-109, a small molecule NE inhibitor, would inhibit CF mucin degradation in vitro. METHODS: Sputa were collected from CF patients (n=5) chronically or intermittently infected with Pseudomonas aeruginosa (P.a.). Mucin degradation was analyzed using western blot. Protease inhibitor studies were performed using alpha1-proteinase inhibitor (A1-PI Prolastin®) and KRP-109. Elastase activity assays were performed using spectrophotometry. RESULTS: There were significant differences in the amount of active NE in different CF sputum samples. KRP-109 decreased the NE driven mucin degradation in vitro. Pseudomonas elastases appeared to blunt elastase inhibition by A1-PI or KRP-109. CONCLUSION: Inhibitors of neutrophil and Pseudomonas-derived elastases might rescue mucus clearance and reverse airway obstruction in CF.
Authors: Oliver W Meldrum; Kylie B R Belchamber; Kiarina D Chichirelo-Konstantynovych; Katie L Horton; Tetyana V Konstantynovych; Merete B Long; Melissa J McDonnell; Lidia Perea; Alberto L Garcia-Basteiro; Michael R Loebinger; Raquel Duarte; Holly R Keir Journal: ERJ Open Res Date: 2022-05-23
Authors: Vincent D Giacalone; Brian S Dobosh; Amit Gaggar; Rabindra Tirouvanziam; Camilla Margaroli Journal: Int J Mol Sci Date: 2020-05-08 Impact factor: 5.923