Improvement of defective cystic fibrosis airway epithelial wound repair after CFTR rescue

Trinh, Nguyen Thu Ngan; Bardou, Olivier; Privé, Anik; Maillé, Émilie; Adam, Damien; Lingee, S.; Ferraro, Pasquale; Desrosiers, Marie-Pierre; Coraux, Christelle; Brochiero, Emmanuelle

doi:10.1183/09031936.00221711

Cited by 80 publications

(124 citation statements)

References 47 publications

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“…Recent data have reported the critical role of the cystic fibrosis transmembrane conductance regulator (Cftr) gene in airway epithelial wound healing (Schiller et al, 2010;Trinh et al, 2012). Although the mechanisms connecting Cftr deficiency to defective klf4 induction and enhanced HAEC activation remain to be established, our results point to a disturbed repair process of CF HAECs characterized by fast but desynchronized initiation of BC activation and proliferation in a primary 3D model of epithelium damage.…”

Section: Discussionmentioning

confidence: 60%

Cx26 regulates proliferation of repairing basal airway epithelial cells

Crespin

Bacchetta

Saab

et al. 2014

The International Journal of Biochemistry & Cell Biology

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The recovery of an intact epithelium following injury is critical for restoration of lung homeostasis, a process that may be altered in cystic fibrosis (CF). In response to injury, progenitor cells in the undamaged areas migrate, proliferate and re-differentiate to regenerate an intact airway epithelium. The mechanisms regulating this regenerative response are, however, not well understood. In a model of circular wound injury of well-differentiated human airway epithelial cell (HAEC) cultures, we identified the gap junction protein Cx26 as an important regulator of cell proliferation. We report that induction of Cx26 in repairing HAECs is associated with cell proliferation. We also show that Cx26 is expressed in a population of CK14-positive basal-like cells. Cx26 silencing in immortalized cell lines using siRNA and in primary HAECs using lentiviral-transduced shRNA enhanced Ki67-labeling index and Ki67 mRNA, indicating that Cx26 acts a negative regulator of HAEC proliferation.Cx26 silencing also markedly decreased the transcription of KLF4 in immortalized HAECs.We further show that CF HAECs exhibited deregulated expression of KLF4, Ki67 and Cx26 as well enhanced rate of wound closure in the early response to injury. These results point to an altered repair process of CF HAECs characterized by rapid but desynchronized initiation of HAEC activation and proliferation.

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Section: Discussionmentioning

confidence: 60%

Cx26 regulates proliferation of repairing basal airway epithelial cells

Crespin

Bacchetta

Saab

et al. 2014

The International Journal of Biochemistry & Cell Biology

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“…Primary human airway cells were isolated from tissues obtained from non-CF and CF patients (with class II mutations, including ΔF508/ ΔF508 and ΔF508/N1303K, median age 23 years, mean forced expiratory volume in 1s of 82±9%) undergoing nasal polypectomy at CHUM (Montreal, QC, Canada) and CHU Sainte-Justine (Montreal, QC, Canada) hospitals [28], according to approved ethical protocols. Written informed consents have been obtained from all participants.…”

Section: Methodsmentioning

confidence: 99%

“…For CFTR [28] and epidermal growth factor receptor (EGFR) [30] detection, the upper section of the membrane was incubated with the polyclonal anti-CFTR 596 antibody (CFFT) or anti-EGFR antibody (anti-erbB1; Cell Signaling Technology, Danvers, MA, USA); whereas the bottom section was incubated with purified mouse anti-β-actin monoclonal antibody (Cedarlane Laboratory, Burlington, ON, Canada) to ensure equivalent loading. The expression of CFTR was reported as percentage relative to cells with LB, after normalisation to β-actin signal.…”

Section: Immunoblottingmentioning

confidence: 99%

“…Short-circuit currents (Isc) [30][31][32] were measured through CFBE and primary airway cells [28] that had been cultured on coated permeant filters until they reached an air-liquid interface and formed a polarised epithelium with high resistance [33,34]. Measurements were performed (see supplementary material for details) on intact monolayers or through apical membranes after permeabilisation of the basolateral side with 7.5 µM amphotericin B (Sigma-Aldrich), in the presence of a symmetrical physiological solution or a chloride (Cl − ) gradient, as indicated in figure legends.…”

Section: Electrophysiologymentioning

confidence: 99%

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Deleterious impact ofPseudomonas aeruginosaon cystic fibrosis transmembrane conductance regulator function and rescue in airway epithelial cells

et al. 2015

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The epithelial response to bacterial airway infection, a common feature of lung diseases such as chronic obstructive pulmonary disease and cystic fibrosis, has been extensively studied. However, its impact on cystic fibrosis transmembrane conductance regulator (CFTR) channel function is not clearly defined. Our aims were, therefore, to evaluate the effect of Pseudomonas aeruginosa on CFTR function and expression in non-cystic fibrosis airway epithelial cells, and to investigate its impact on ΔF508-CFTR rescue by the VRT-325 corrector in cystic fibrosis cells.CFTR expression/maturation was evaluated by immunoblotting and its function by short-circuit current measurements.A 24-h exposure to P. aeruginosa diffusible material (PsaDM) reduced CFTR currents as well as total and membrane protein expression of the wildtype (wt) CFTR protein in CFBE-wt cells. In CFBE-ΔF508 cells, PsaDM severely reduced CFTR maturation and current rescue induced by VRT-325. We also confirmed a deleterious impact of PsaDM on wt-CFTR currents in non-cystic fibrosis primary airway cells as well as on the rescue of ΔF508-CFTR function induced by VRT-325 in primary cystic fibrosis cells.These findings show that CFTR function could be impaired in non-cystic fibrosis patients infected by P. aeruginosa. Our data also suggest that CFTR corrector efficiency may be affected by infectious components, which should be taken into account in screening assays of correctors. @ERSpublications Exposure of airway epithelial cell to P. aeruginosa impairs CFTR function, expression and rescue by correctors http://ow.ly/IleTw

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“…In bronchial tissues, CFTR protein is expressed in the submucosal glands and the apical surface of ciliated epithelial cells [27,28], and has been shown to play a critical role in epithelial wound repair [29]. Normally, effective mucociliary transport is facilitated by sufficient hydration of the airway surface liquid (ASL); an important periciliary and mucus layer that lines the airway tract [30,31].…”

Section: Airwaysmentioning

confidence: 99%

Targeting a genetic defect: cystic fibrosis transmembrane conductance regulator modulators in cystic fibrosis

Derichs

2013

Eur Respir Rev

105

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Cystic fibrosis (CF) is caused by genetic mutations that affect the cystic fibrosis transmembrane conductance regulator (CFTR) protein. These mutations can impact the synthesis and transfer of the CFTR protein to the apical membrane of epithelial cells, as well as influencing the gating or conductance of chloride and bicarbonate ions through the channel. CFTR dysfunction results in ionic imbalance of epithelial secretions in several organ systems, such as the pancreas, gastrointestinal tract, liver and the respiratory system. Since discovery of the CFTR gene in 1989, research has focussed on targeting the underlying genetic defect to identify a disease-modifying treatment for CF. Investigated management strategies have included gene therapy and the development of small molecules that target CFTR mutations, known as CFTR modulators. CFTR modulators are typically identified by high-throughput screening assays, followed by preclinical validation using cell culture systems. Recently, one such modulator, the CFTR potentiator ivacaftor, was approved as an oral therapy for CF patients with the G551D-CFTR mutation. The clinical development of ivacaftor not only represents a breakthrough in CF care but also serves as a noteworthy example of personalised medicine.

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Improvement of defective cystic fibrosis airway epithelial wound repair after CFTR rescue

Cited by 80 publications

References 47 publications

Cx26 regulates proliferation of repairing basal airway epithelial cells

Cx26 regulates proliferation of repairing basal airway epithelial cells

Deleterious impact ofPseudomonas aeruginosaon cystic fibrosis transmembrane conductance regulator function and rescue in airway epithelial cells

Targeting a genetic defect: cystic fibrosis transmembrane conductance regulator modulators in cystic fibrosis

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