{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T09:36:28Z","timestamp":1790501788974,"version":"4.1.0"},"reference-count":42,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2020,2,19]],"date-time":"2020-02-19T00:00:00Z","timestamp":1582070400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000082","name":"Cystic Fibrosis Canada","doi-asserted-by":"publisher","award":["3172"],"award-info":[{"award-number":["3172"]}],"id":[{"id":"10.13039\/501100000082","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000082","name":"Cystic Fibrosis Canada","doi-asserted-by":"publisher","award":["480991"],"award-info":[{"award-number":["480991"]}],"id":[{"id":"10.13039\/501100000082","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["CIHR MOP-125855"],"award-info":[{"award-number":["CIHR MOP-125855"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["RGPIN-2016-05577"],"award-info":[{"award-number":["RGPIN-2016-05577"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Chronic infection and inflammation are the primary causes of declining lung function in Cystic Fibrosis (CF) patients. ORKAMBI\u00ae (Lumacaftor-Ivacaftor) is an approved combination therapy for Cystic Fibrosis (CF) patients bearing the most common mutation, F508del, in the cystic fibrosis conductance regulator (CFTR) protein. It has been previously shown that ORKAMBI\u00ae-mediated rescue of CFTR is reduced by a pre-existing Pseudomonas aeruginosa infection. Here, we show that the infection of F508del-CFTR human bronchial epithelial (HBE) cells with lab strain and four different clinical strains of P. aeruginosa, isolated from the lung sputum of CF patients, decreases CFTR function in a strain-specific manner by 48 to 88%. The treatment of infected cells with antibiotic tobramycin or cationic antimicrobial peptide 6K-F17 was found to decrease clinical strain bacterial growth on HBE cells and restore ORKAMBI\u00ae-mediated rescue of F508del-CFTR function. Further, 6K-F17 was found to downregulate the expression of pro-inflammatory cytokines, interleukin (IL)-8, IL-6, and tumor necrosis factor-\u03b1 in infected HBE cells. The results provide strong evidence for a combination therapy approach involving CFTR modulators and anti-infectives (i.e., tobramycin and\/or 6K-F17) to improve their overall efficacy in CF patients.<\/jats:p>","DOI":"10.3390\/biom10020334","type":"journal-article","created":{"date-parts":[[2020,2,21]],"date-time":"2020-02-21T10:49:16Z","timestamp":1582282156000},"page":"334","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":35,"title":["Anti-Infectives Restore ORKAMBI\u00ae Rescue of F508del-CFTR Function in Human Bronchial Epithelial Cells Infected with Clinical Strains of P. aeruginosa"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0237-4079","authenticated-orcid":false,"given":"Onofrio","family":"Laselva","sequence":"first","affiliation":[{"name":"Division of Molecular Medicine, Research Institute, Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"},{"name":"Department of Physiology, University of Toronto, Toronto, ON M5S 1A8, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5689-5232","authenticated-orcid":false,"given":"Tracy A.","family":"Stone","sequence":"additional","affiliation":[{"name":"Division of Molecular Medicine, Research Institute, Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"},{"name":"Department of Biochemistry, University of Toronto, Toronto, ON M5S 1A8, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christine E.","family":"Bear","sequence":"additional","affiliation":[{"name":"Division of Molecular Medicine, Research Institute, Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"},{"name":"Department of Physiology, University of Toronto, Toronto, ON M5S 1A8, Canada"},{"name":"Department of Biochemistry, University of Toronto, Toronto, ON M5S 1A8, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles M.","family":"Deber","sequence":"additional","affiliation":[{"name":"Division of Molecular Medicine, Research Institute, Hospital for Sick Children, Toronto, ON M5G 0A4, Canada"},{"name":"Department of Biochemistry, University of Toronto, Toronto, ON M5S 1A8, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,2,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1126\/science.2475911","article-title":"Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA","volume":"245","author":"Riordan","year":"1989","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"809","DOI":"10.1016\/0092-8674(92)90155-6","article-title":"Purification and functional reconstitution of the cystic fibrosis transmembrane conductance regulator (CFTR)","volume":"68","author":"Bear","year":"1992","journal-title":"Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1992","DOI":"10.1056\/NEJMra043184","article-title":"Cystic Fibrosis","volume":"352","author":"Rowe","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1136\/thoraxjnl-2013-203204","article-title":"Airway inflammation in cystic fibrosis: molecular mechanisms and clinical implications","volume":"68","author":"Kerem","year":"2013","journal-title":"Thorax"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"351","DOI":"10.1056\/NEJMra1300109","article-title":"Origins of Cystic Fibrosis Lung Disease","volume":"372","author":"Stoltz","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1080\/17476348.2016.1177460","article-title":"Pseudomonas aeruginosa infection in cystic fibrosis: pathophysiological mechanisms and therapeutic approaches","volume":"10","author":"Turnbull","year":"2016","journal-title":"Expert Rev. Respir. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1007\/s40291-018-0372-6","article-title":"Innovative Therapies for Cystic Fibrosis: The Road from Treatment to Cure","volume":"23","author":"Cabrini","year":"2019","journal-title":"Mol. Diagn. Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"6822","DOI":"10.1128\/IAI.73.10.6822-6830.2005","article-title":"Influence of Cystic Fibrosis Transmembrane Conductance Regulator on Gene Expression in Response to Pseudomonas aeruginosa Infection of Human Bronchial Epithelial Cells","volume":"73","author":"Reiniger","year":"2005","journal-title":"Infect. Immun."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Bhagirath, A.Y., Li, Y., Somayajula, D., Dadashi, M., Badr, S., and Duan, K. (2016). Cystic fibrosis lung environment and Pseudomonas aeruginosa infection. BMC Pulm. Med., 16.","DOI":"10.1186\/s12890-016-0339-5"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.molmed.2011.10.003","article-title":"CFTR: folding, misfolding and correcting the \u0394F508 conformational defect","volume":"18","author":"Lukacs","year":"2012","journal-title":"Trends Mol. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"18843","DOI":"10.1073\/pnas.1105787108","article-title":"Correction of the F508del-CFTR protein processing defect in vitro by the investigational drug VX-809","volume":"108","author":"Hadida","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1783","DOI":"10.1056\/NEJMoa1409547","article-title":"Lumacaftor\u2013Ivacaftor in Patients with Cystic Fibrosis Homozygous for Phe508del CFTR","volume":"373","author":"Wainwright","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1586\/17476348.2016.1122527","article-title":"Lumacaftor alone and combined with ivacaftor: preclinical and clinical trial experience of F508del CFTR correction","volume":"10","author":"Brewington","year":"2016","journal-title":"Expert Rev. Respir. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"231","DOI":"10.2217\/fmb.14.107","article-title":"Small colony variants of Pseudomonas aeruginosa in chronic bacterial infection of the lung in cystic fibrosis","volume":"10","author":"Evans","year":"2015","journal-title":"Future Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/j.tim.2016.01.008","article-title":"Pseudomonas aeruginosa Evolutionary Adaptation and Diversification in Cystic Fibrosis Chronic Lung Infections","volume":"24","author":"Winstanley","year":"2016","journal-title":"Trends Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1590","DOI":"10.1183\/09031936.00076214","article-title":"Deleterious impact of Pseudomonas aeruginosa on cystic fibrosis transmembrane conductance regulator function and rescue in airway epithelial cells","volume":"45","author":"Trinh","year":"2015","journal-title":"Eur. Respir. J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"470","DOI":"10.3389\/fcimb.2017.00470","article-title":"Quorum Sensing Down-Regulation Counteracts the Negative Impact of Pseudomonas aeruginosa on CFTR Channel Expression, Function and Rescue in Human Airway Epithelial Cells","volume":"7","author":"Ruffin","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Stanton, B.A., Coutermarsh, B., Barnaby, R., and Hogan, D. (2015). Pseudomonas aeruginosa Reduces VX-809 Stimulated F508del-CFTR Chloride Secretion by Airway Epithelial Cells. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0127742"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1002\/psc.2674","article-title":"Antibacterial and anti-inflammatory activity of a temporin B peptide analogue on an in vitro model of cystic fibrosis","volume":"20","author":"Bezzerri","year":"2014","journal-title":"J. Pept. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"L48","DOI":"10.1152\/ajplung.00305.2013","article-title":"Trimethylangelicin promotes the functional rescue of mutant F508del CFTR protein in cystic fibrosis airway cells","volume":"307","author":"Favia","year":"2014","journal-title":"Am. J. Physiol. Cell. Mol. Physiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"L380","DOI":"10.1152\/ajplung.00129.2010","article-title":"Trimethylangelicin reduces IL-8 transcription and potentiates CFTR function","volume":"300","author":"Tamanini","year":"2011","journal-title":"Am. J. Physiol. Cell. Mol. Physiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"719","DOI":"10.3389\/fphar.2018.00719","article-title":"Molecular Mechanism of Action of Trimethylangelicin Derivatives as CFTR Modulators","volume":"9","author":"LaSelva","year":"2018","journal-title":"Front. Pharmacol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/j.bcp.2016.09.005","article-title":"The investigational Cystic Fibrosis drug Trimethylangelicin directly modulates CFTR by stabilizing the first membrane-spanning domain","volume":"119","author":"LaSelva","year":"2016","journal-title":"Biochem. Pharmacol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"612","DOI":"10.1124\/mol.118.111799","article-title":"Correctors of the Major Cystic Fibrosis Mutant Interact through Membrane-Spanning Domains","volume":"93","author":"LaSelva","year":"2018","journal-title":"Mol. Pharmacol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1517\/14740330903036083","article-title":"Inhaled therapeutics for prevention and treatment of pneumonia","volume":"8","author":"Safdar","year":"2009","journal-title":"Expert Opin. Drug Saf."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"806","DOI":"10.1016\/j.jcf.2013.02.007","article-title":"Tobramycin is a suppressor of premature termination codons","volume":"12","author":"Altamura","year":"2013","journal-title":"J. Cyst. Fibros."},{"key":"ref_27","first-page":"194","article-title":"Antimicrobial Peptides: An Emerging Category of Therapeutic Agents","volume":"6","author":"Mahlapuu","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.cbpa.2017.03.014","article-title":"Host defense antimicrobial peptides as antibiotics: design and application strategies","volume":"38","author":"Mishra","year":"2017","journal-title":"Curr. Opin. Chem. Boil."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"D590","DOI":"10.1093\/nar\/gkh025","article-title":"APD: The Antimicrobial Peptide Database","volume":"32","author":"Wang","year":"2004","journal-title":"Nucleic Acids Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"33960","DOI":"10.1074\/jbc.M507042200","article-title":"Basis for selectivity of cationic antimicrobial peptides for bacterial versus mammalian membranes","volume":"280","author":"Glukhov","year":"2005","journal-title":"J. Boil. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6276","DOI":"10.1021\/acs.jmedchem.9b00657","article-title":"Positive Charge Patterning and Hydrophobicity of Membrane-Active Antimicrobial Peptides as Determinants of Activity, Toxicity, and Pharmacokinetic Stability","volume":"62","author":"Stone","year":"2019","journal-title":"J. Med. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"14728","DOI":"10.1038\/s41598-018-33016-7","article-title":"Activity of a novel antimicrobial peptide against Pseudomonas aeruginosa biofilms","volume":"8","author":"Beaudoin","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"476","DOI":"10.1016\/j.jcf.2018.12.001","article-title":"Isogenic cell models of cystic fibrosis-causing variants in natively expressing pulmonary epithelial cells","volume":"18","author":"Valley","year":"2019","journal-title":"J. Cyst. Fibros."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"LaSelva, O., Eckford, P.D., Bartlett, C., Ouyang, H., Na Gunawardena, T., Gonska, T., Moraes, T., and Bear, C.E. (2019). Functional rescue of c.3846G>A (W1282X) in patient-derived nasal cultures achieved by inhibition of nonsense mediated decay and protein modulators with complementary mechanisms of action. J. Cyst. Fibros.","DOI":"10.1016\/j.jcf.2019.12.001"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1124\/mol.119.117143","article-title":"ORKAMBI-Mediated Rescue of Mucociliary Clearance in Cystic Fibrosis Primary Respiratory Cultures Is Enhanced by Arginine Uptake, Arginase Inhibition, and Promotion of Nitric Oxide Signaling to the Cystic Fibrosis Transmembrane Conductance Regulator Channel","volume":"96","author":"Wu","year":"2019","journal-title":"Mol. Pharmacol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1224","DOI":"10.15252\/emmm.201607137","article-title":"Orkambi\u00ae and amplifier co-therapy improves function from a rare CFTR mutation in gene-edited cells and patient tissue","volume":"9","author":"Molinski","year":"2017","journal-title":"EMBO Mol. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"549","DOI":"10.4049\/jimmunol.164.2.549","article-title":"Cutting edge: cationic antimicrobial peptides block the binding of lipopolysaccharide (LPS) to LPS binding protein","volume":"164","author":"Scott","year":"2000","journal-title":"J. Immunol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Bomberger, J.M., Ye, S., MacEachran, D.P., Koeppen, K., Barnaby, R.L., O\u2019Toole, G.A., and Stanton, B.A. (2011). A Pseudomonas aeruginosa Toxin that Hijacks the Host Ubiquitin Proteolytic System. PLoS Pathog., 7.","DOI":"10.1371\/journal.ppat.1001325"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"C357","DOI":"10.1152\/ajpcell.00373.2016","article-title":"Effects of Pseudomonas aeruginosa on CFTR chloride secretion and the host immune response","volume":"312","author":"Stanton","year":"2017","journal-title":"Am. J. Physiol. Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1038\/nrm1949","article-title":"New insights into cystic fibrosis: molecular switches that regulate CFTR","volume":"7","author":"Guggino","year":"2006","journal-title":"Nat. Rev. Mol. Cell Boil."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1038\/nrmicro1441","article-title":"The co-evolution of host cationic antimicrobial peptides and microbial resistance","volume":"4","author":"Peschel","year":"2006","journal-title":"Nat. Rev. Genet."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1621","DOI":"10.1021\/ci200583t","article-title":"Testing Physical Models of Passive Membrane Permeation","volume":"52","author":"Leung","year":"2012","journal-title":"J. Chem. Inf. Model."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/10\/2\/334\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T08:59:03Z","timestamp":1760173143000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/10\/2\/334"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,2,19]]},"references-count":42,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2020,2]]}},"alternative-id":["biom10020334"],"URL":"https:\/\/doi.org\/10.3390\/biom10020334","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,2,19]]}}}