Claudio Sorio1,2, Alessio Montresor1, Matteo Bolomini-Vittori1,2,3, Sara Caldrer1,2, Barbara Rossi1, Silvia Dusi1, Stefano Angiari1, Jan E Johansson1,2, Marzia Vezzalini1,2, Teresinha Leal4, Elisa Calcaterra1,2, Baroukh M Assael5, Paola Melotti5, Carlo Laudanna1. 1. 1 Division of General Pathology, Department of Medicine, and. 2. 2 Cystic Fibrosis Translational Research Laboratory "Daniele Lissandrini," Department of Medicine, University of Verona School of Medicine, Verona, Italy. 3. 3 Department of Tumor Immunology (278), Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, the Netherlands. 4. 4 Faculté de Pharmacie et des Sciences Biomédicales, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Brussels, Belgium; and. 5. 5 Centro Fibrosi Cistica, Azienda Ospedaliera Universitaria Integrata di Verona, Verona, Italy.
Abstract
RATIONALE: Cystic fibrosis (CF) is a common genetic disease caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Persistent lung inflammation, characterized by increasing polymorphonuclear leukocyte recruitment, is a major cause of the decline in respiratory function in patients with CF and is a leading cause of morbidity and mortality. CFTR is expressed in various cell types, including leukocytes, but its involvement in the regulation of leukocyte recruitment is unknown. OBJECTIVES: We evaluated whether CF leukocytes might present with alterations in cell adhesion and migration, a key process governing innate and acquired immune responses. METHODS: We used ex vivo adhesion and chemotaxis assays, flow cytometry, immunofluorescence, and GTPase activity assays in this study. MEASUREMENTS AND MAIN RESULTS: We found that chemoattractant-induced activation of β1 and β2 integrins and of chemotaxis is defective in mononuclear cells isolated from patients with CF. In contrast, polymorphonuclear leukocyte adhesion and chemotaxis were normal. The functionality of β1 and β2 integrins was restored by treatment of CF monocytes with the CFTR-correcting drugs VRT325 and VX809. Moreover, treatment of healthy monocytes with the CFTR inhibitor CFTR(inh)-172 blocked integrin activation by chemoattractants. In a murine model of lung inflammation, we found that integrin-independent migration of CF monocytes into the lung parenchyma was normal, whereas, in contrast, integrin-dependent transmigration into the alveolar space was impaired. Finally, signal transduction analysis showed that, in CF monocytes, chemoattractant-triggered activation of RhoA and CDC42 Rho small GTPases (controlling integrin activation and chemotaxis, respectively) was strongly deficient. CONCLUSIONS: Altogether, these data highlight the critical regulatory role of CFTR in integrin activation by chemoattractants in monocytes and identify CF as a new, cell type-selective leukocyte adhesion deficiency disease, providing new insights into CF pathogenesis.
RATIONALE: Cystic fibrosis (CF) is a common genetic disease caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Persistent lung inflammation, characterized by increasing polymorphonuclear leukocyte recruitment, is a major cause of the decline in respiratory function in patients with CF and is a leading cause of morbidity and mortality. CFTR is expressed in various cell types, including leukocytes, but its involvement in the regulation of leukocyte recruitment is unknown. OBJECTIVES: We evaluated whether CF leukocytes might present with alterations in cell adhesion and migration, a key process governing innate and acquired immune responses. METHODS: We used ex vivo adhesion and chemotaxis assays, flow cytometry, immunofluorescence, and GTPase activity assays in this study. MEASUREMENTS AND MAIN RESULTS: We found that chemoattractant-induced activation of β1 and β2 integrins and of chemotaxis is defective in mononuclear cells isolated from patients with CF. In contrast, polymorphonuclear leukocyte adhesion and chemotaxis were normal. The functionality of β1 and β2 integrins was restored by treatment of CF monocytes with the CFTR-correcting drugs VRT325 and VX809. Moreover, treatment of healthy monocytes with the CFTR inhibitor CFTR(inh)-172 blocked integrin activation by chemoattractants. In a murine model of lung inflammation, we found that integrin-independent migration of CF monocytes into the lung parenchyma was normal, whereas, in contrast, integrin-dependent transmigration into the alveolar space was impaired. Finally, signal transduction analysis showed that, in CF monocytes, chemoattractant-triggered activation of RhoA and CDC42 Rho small GTPases (controlling integrin activation and chemotaxis, respectively) was strongly deficient. CONCLUSIONS: Altogether, these data highlight the critical regulatory role of CFTR in integrin activation by chemoattractants in monocytes and identify CF as a new, cell type-selective leukocyte adhesion deficiency disease, providing new insights into CF pathogenesis.
Authors: Jonas C Schupp; Sara Khanal; Jose L Gomez; Maor Sauler; Taylor S Adams; Geoffrey L Chupp; Xiting Yan; Sergio Poli; Yujiao Zhao; Ruth R Montgomery; Ivan O Rosas; Charles S Dela Cruz; Emanuela M Bruscia; Marie E Egan; Naftali Kaminski; Clemente J Britto Journal: Am J Respir Crit Care Med Date: 2020-11-15 Impact factor: 21.405
Authors: Charles R Esther; Marianne S Muhlebach; Camille Ehre; David B Hill; Matthew C Wolfgang; Mehmet Kesimer; Kathryn A Ramsey; Matthew R Markovetz; Ian C Garbarine; M Gregory Forest; Ian Seim; Bryan Zorn; Cameron B Morrison; Martial F Delion; William R Thelin; Diane Villalon; Juan R Sabater; Lidija Turkovic; Sarath Ranganathan; Stephen M Stick; Richard C Boucher Journal: Sci Transl Med Date: 2019-04-03 Impact factor: 17.956
Authors: Zhichao Fan; Elise Pitmon; Lai Wen; Jacqueline Miller; Erik Ehinger; Rana Herro; Wei Liu; Ju Chen; Zbigniew Mikulski; Douglas J Conrad; Alex Marki; Marco Orecchioni; Puja Kumari; Yanfang Peipei Zhu; Paola M Marcovecchio; Catherine C Hedrick; Craig A Hodges; Vijay A Rathinam; Kepeng Wang; Klaus Ley Journal: J Immunol Date: 2022-01-14 Impact factor: 5.422