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Src kinase inhibition reduces inflammatory and cytoskeletal changes in ΔF508 human cholangiocytes and improves cystic fibrosis transmembrane conductance regulator correctors efficacy.
- Source :
-
Hepatology (Baltimore, Md.) [Hepatology] 2018 Mar; Vol. 67 (3), pp. 972-988. Date of Electronic Publication: 2018 Jan 26. - Publication Year :
- 2018
-
Abstract
- Cystic fibrosis transmembrane conductance regulator (CFTR), the channel mutated in cystic fibrosis (CF), is expressed by the biliary epithelium (i.e., cholangiocytes) of the liver. Progressive clinical liver disease (CF-associated liver disease; CFLD) occurs in around 10% of CF patients and represents the third leading cause of death. Impaired secretion and inflammation contribute to CFLD; however, the lack of human-derived experimental models has hampered the understanding of CFLD pathophysiology and the search for a cure. We have investigated the cellular mechanisms altered in human CF cholangiocytes using induced pluripotent stem cells (iPSCs) derived from healthy controls and a ΔF508 CFTR patient. We have devised a novel protocol for the differentiation of human iPSC into polarized monolayers of cholangiocytes. Our results show that iPSC-cholangiocytes reproduced the polarity and the secretory function of the biliary epithelium. Protein kinase A/cAMP-mediated fluid secretion was impaired in ΔF508 cholangiocytes and negligibly improved by VX-770 and VX-809, two small molecule drugs used to correct and potentiate ΔF508 CFTR. Moreover, ΔF508 cholangiocytes showed increased phosphorylation of Src kinase and Toll-like receptor 4 and proinflammatory changes, including increased nuclear factor kappa-light-chain-enhancer of activated B cells activation, secretion of proinflammatory chemokines (i.e., monocyte chemotactic protein 1 and interleukin-8), as well as alterations of the F-actin cytoskeleton. Treatment with Src inhibitor (4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyramidine) decreased the inflammatory changes and improved cytoskeletal defects. Inhibition of Src, along with administration of VX-770 and VX-809, successfully restored fluid secretion to normal levels.<br />Conclusion: Our findings have strong translational potential and indicate that targeting Src kinase and decreasing inflammation may increase the efficacy of pharmacological therapies aimed at correcting the basic ΔF508 defect in CF liver patients. These studies also demonstrate the promise of applying iPSC technology in modeling human cholangiopathies. (Hepatology 2018;67:972-988).<br /> (© 2017 by the American Association for the Study of Liver Diseases.)
- Subjects :
- Animals
Biliary Tract cytology
Biliary Tract drug effects
Biliary Tract pathology
Cell Culture Techniques
Cystic Fibrosis drug therapy
Cystic Fibrosis Transmembrane Conductance Regulator drug effects
Cytokines metabolism
Cytoskeleton metabolism
Epithelial Cells metabolism
Fluorescent Antibody Technique
Humans
Induced Pluripotent Stem Cells drug effects
Induced Pluripotent Stem Cells metabolism
Induced Pluripotent Stem Cells physiology
Inflammation metabolism
Mice
Microscopy, Confocal
Signal Transduction
src-Family Kinases antagonists & inhibitors
Aminophenols pharmacology
Aminopyridines pharmacology
Benzodioxoles pharmacology
Chloride Channel Agonists pharmacology
Cystic Fibrosis physiopathology
Pyrimidines pharmacology
Quinolones pharmacology
src-Family Kinases metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1527-3350
- Volume :
- 67
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Hepatology (Baltimore, Md.)
- Publication Type :
- Academic Journal
- Accession number :
- 28836688
- Full Text :
- https://doi.org/10.1002/hep.29400