Mechanical Properties of Human Bronchial Epithelial Cells Expressing Wt- and Mutant CFTR

Carapeto, Ana P.; Vitorino, Miguel V.; Santos, João Dourado; Ramalho, Sofia; Robalo, Tiago T.; Rodrigues, Mário S.; Farinha, Carlos M.

doi:10.3390/ijms21082916

Cited by 9 publications

(10 citation statements)

References 32 publications

(36 reference statements)

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“…CF-relevant immortalized bronchial epithelial cell lines, CFBE41o- (Cystic Fibrosis bronchial epithelial) cells, stably overexpressing wt- and F508del–CFTR [ 16 ], were used in this work and grown as previously described [ 17 ]. To achieve polarization, cells were seeded on collagen IV pre-coated transwell permeable supports.…”

Section: Methodsmentioning

confidence: 99%

KLF4 Acts as a wt-CFTR Suppressor through an AKT-Mediated Pathway

Sousa

Pankonien

Clarke

et al. 2020

Cells

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Cystic Fibrosis (CF) is caused by >2000 mutations in the CF transmembrane conductance regulator (CFTR) gene, but one mutation—F508del—occurs in ~80% of patients worldwide. Besides its main function as an anion channel, the CFTR protein has been implicated in epithelial differentiation, tissue regeneration, and, when dysfunctional, cancer. However, the mechanisms that regulate such relationships are not fully elucidated. Krüppel-like factors (KLFs) are a family of transcription factors (TFs) playing central roles in development, stem cell differentiation, and proliferation. Herein, we hypothesized that these TFs might have an impact on CFTR expression and function, being its missing link to differentiation. Our results indicate that KLF4 (but not KLF2 nor KLF5) is upregulated in CF vs. non-CF cells and that it negatively regulates wt-CFTR expression and function. Of note, F508del–CFTR expressing cells are insensitive to KLF4 modulation. Next, we investigated which KLF4-related pathways have an effect on CFTR. Our data also show that KLF4 modulates wt-CFTR (but not F508del–CFTR) via both the serine/threonine kinase AKT1 (AKT) and glycogen synthase kinase 3 beta (GSK3β) signaling. While AKT acts positively, GSK3β is a negative regulator of CFTR. This crosstalk between wt-CFTR and KLF4 via AKT/ GSK3β signaling, which is disrupted in CF, constitutes a novel mechanism linking CFTR to the epithelial differentiation.

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Section: Methodsmentioning

confidence: 99%

KLF4 Acts as a wt-CFTR Suppressor through an AKT-Mediated Pathway

Sousa

Pankonien

Clarke

et al. 2020

Cells

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“…This results in an alteration of cellular and matrix stiffness. Human epithelial cells derived from patients’ airways with CF and CFTR mutant cells have been found to have a lower Young’s modulus than normal human epithelial cells [ 65 , 66 ]. Alveolar matrix remodeling and fibrosis is present in the CF lung and leads to stiffening of alveolar tissues.…”

Section: The Role Of Collagen In Airway Disease and Disease-associmentioning

confidence: 99%

Role of Collagen in Airway Mechanics

et al. 2021

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Collagen is the most abundant airway extracellular matrix component and is the primary determinant of mechanical airway properties. Abnormal airway collagen deposition is associated with the pathogenesis and progression of airway disease. Thus, understanding how collagen affects healthy airway tissue mechanics is essential. The impact of abnormal collagen deposition and tissue stiffness has been an area of interest in pulmonary diseases such as cystic fibrosis, asthma, and chronic obstructive pulmonary disease. In this review, we discuss (1) the role of collagen in airway mechanics, (2) macro- and micro-scale approaches to quantify airway mechanics, and (3) pathologic changes associated with collagen deposition in airway diseases. These studies provide important insights into the role of collagen in airway mechanics. We summarize their achievements and seek to provide biomechanical clues for targeted therapies and regenerative medicine to treat airway pathology and address airway defects.

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“…Thus, CFTR membrane retention, CFTR association with NHERF-1 and cytoskeleton organization are key factors in CF disease mechanisms. As a way to study actin cytoskeleton alteration in CF cells, Carapeto et al [ 18 ] used Atomic Force Microscopy and Force Feedback Microscopy to investigate the mechanical properties of cystic fibrosis bronchial epithelial (CFBE) cells stably transduced with either wild type (wt-) or F508del-CFTR. They found that the expression of mutant CFTR causes a decrease in the cell’s apparent Young (elastic) modulus as compared to the expression of the wt protein [ 18 ].…”

mentioning

confidence: 99%

“…As a way to study actin cytoskeleton alteration in CF cells, Carapeto et al [ 18 ] used Atomic Force Microscopy and Force Feedback Microscopy to investigate the mechanical properties of cystic fibrosis bronchial epithelial (CFBE) cells stably transduced with either wild type (wt-) or F508del-CFTR. They found that the expression of mutant CFTR causes a decrease in the cell’s apparent Young (elastic) modulus as compared to the expression of the wt protein [ 18 ]. This decrease is likely the consequence of a more disorganized actin cytoskeleton possibly due to a depolymerization of the actin fibers network below the cell membrane, related with the F508del mutation [ 18 ].…”

mentioning

confidence: 99%

“…They found that the expression of mutant CFTR causes a decrease in the cell’s apparent Young (elastic) modulus as compared to the expression of the wt protein [ 18 ]. This decrease is likely the consequence of a more disorganized actin cytoskeleton possibly due to a depolymerization of the actin fibers network below the cell membrane, related with the F508del mutation [ 18 ].…”

mentioning

confidence: 99%

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