Identification and Function of Cyclic Nucleotide Phosphodiesterase Isoenzymes in Airway Epithelial Cells

Trikafta, currently the leading therapeutic in Cystic Fibrosis (CF), has demonstrated a real clinical benefit. This treatment is the triple combination therapy of two folding correctors elexacaftor/tezacaftor (VX445/VX661) plus the gating potentiator ivacaftor (VX770). In this study, our aim was to compare the properties of F508del-CFTR in cells treated with either lumacaftor (VX809), tezacaftor, elexacaftor, elexacaftor/tezacaftor with or without ivacaftor. We studied F508del-CFTR function, maturation and membrane localisation by Ussing chamber and whole-cell patch clamp recordings, Western blot and immunolocalization experiments. With human primary airway epithelial cells and the cell lines CFBE and BHK expressing F508del, we found that, whereas the combination elexacaftor/tezacaftor/ivacaftor was efficient in rescuing F508del-CFTR abnormal maturation, apical membrane location and function, the presence of ivacaftor limits these effects. The basal F508del-CFTR short-circuit current was significantly increased by elexacaftor/tezacaftor/ivacaftor and elexacaftor/tezacaftor compared to other correctors and non-treated cells, an effect dependent on ivacaftor and cAMP. These results suggest that the level of the basal F508del-CFTR current might be a marker for correction efficacy in CF cells. When cells were treated with ivacaftor combined to any correctors, the F508del-CFTR current was unresponsive to the subsequently acute addition of ivacaftor unlike the CFTR potentiators genistein and Cact-A1 which increased elexacaftor/tezacaftor/ivacaftor and elexacaftor/tezacaftor-corrected F508del-CFTR currents. These findings show that ivacaftor reduces the correction efficacy of Trikafta. Thus, combining elexacaftor/tezacaftor with a different potentiator might improve the therapeutic efficacy for treating CF patients.

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Regulation of TLR2 Expression and Function in Human Airway Epithelial Cells

The Journal of Membrane Biology ◽

10.1007/s00232-009-9175-3 ◽

2009 ◽

Vol 229 (2) ◽

pp. 101-113 ◽

Cited By ~ 24

Author(s):

Tamene Melkamu ◽

Diane Squillace ◽

Hirohito Kita ◽

Scott M. O’Grady

Keyword(s):

Epithelial Cells ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Tlr2 Expression ◽

Human Airway ◽

And Function ◽

Human Airway Epithelial Cells

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Expression of IL-4/IL-13 receptors in differentiating human airway epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00422.2009 ◽

2010 ◽

Vol 299 (5) ◽

pp. L681-L693 ◽

Cited By ~ 16

Author(s):

Steven R. White ◽

Linda D. Martin ◽

Randi Stern ◽

Bharathi Laxman ◽

Bertha A. Marroquin

Keyword(s):

Epithelial Cells ◽

Airway Inflammation ◽

Ciliated Cell ◽

Airway Epithelial Cells ◽

Receptor Expression ◽

Airway Epithelial ◽

Functional Responses ◽

Receptor Complexes ◽

Differentiated Cells ◽

And Function

IL-4 and IL-13 elicit several important responses in airway epithelium including chemokine secretion and mucous secretion that may contribute to airway inflammation, cell migration, and differentiation. These cytokines have overlapping but not identical effector profiles likely due to shared subunits in their receptor complexes. These receptors are variably described in epithelial cells, and the relative expression, localization, and function of these receptors in differentiated and repairing epithelial cells are not clear. We examined IL-4/IL-13 receptor expression and localization in primary airway epithelial cells collected from normal human lungs and grown under conditions yielding both undifferentiated and differentiated cells inclusive of basal, goblet, and ciliated cell phenotypes. Gene expression of the IL-4Rα, IL-2Rγc, IL-13Rα1, and IL-13Rα2 receptor subunits increased with differentiation, but different patterns of localization and protein abundance were seen for each subunit based on both differentiation and the cell subtypes present. Increased expression of receptor subunits observed in more differentiated cells was associated with more substantial functional responses to IL-4 stimulation including increased eotaxin-3 expression and accelerated migration after injury. We demonstrate substantial differences in IL-4/IL-13 receptor subunit expression and responsiveness to IL-4 based on the extent of airway epithelial cell differentiation and suggest that these differences may have functional consequences in airway inflammation.

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ID2008 Aerosol-based cell delivery as an innovative treatment for lung diseases

Biomedical Research and Therapy ◽

10.15419/bmrat.v4is.251 ◽

2017 ◽

Vol 4 (S) ◽

pp. 41 ◽

Cited By ~ 1

Author(s):

Badrul Hisham Yahaya

Keyword(s):

Epithelial Cells ◽

Cell Therapy ◽

Airway Epithelial Cells ◽

Cell Delivery ◽

Airway Epithelial ◽

Promising Alternative ◽

Airway Injury ◽

Lung Injuries ◽

And Function

Endogenous repair mechanism of airway epithelial cells often fails to achieve sufficient cellular turnover and diminish with age, thus leading to permanent alterations in the structure and function of the airway epithelium. The therapeutic use of stem cells and progenitor cells represents a promising alternative for clinical strategy in treating acute and chronic lung disorders. Aerosol-based cell therapy is a novel therapeutic strategy in enhancing reparative process following both acute and chronic lung injuries. In such background, this study was aimed to determine the effect of aerosol-based cell delivery using MicroSprayer® Aerosolizer in the setting of acute lung injury (ALI) and ovalbumin-induced airway injury in the rabbit. In vitro evaluation revealed that the aerosol technique didn’t cause a significant effect on cell morphology, viability and proliferation capability over the course of cell culture period. Aerosol delivery of airway epithelial cells (AEC) and mesenchymal stem cell (MSC) resulted in uniform distribution in the distal airway and lung interstitial region. Short term assessment showed that cells delivered to the lungs via aerosol was found to be safe for transplantation with no signs of cell rejection and histopathological alterations in the liver and spleen of all treated animals. Histological evidences also demonstrated that administration of AEC and MSC via aerosolization into the respiratory airway prevented lung inflammation as well as resulted in improvement of both alveolar damage and permeability. To our knowledge, this is the first report of aerosol cell delivery via a Microsprayer® Aerosolizer device to the lungs of rabbits to treat airway injuries. Our findings provide a promising evidence that aerosol-based cell therapy may provide a basis for the development of an innovative approach for the treatment lung injuries.

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Double-stranded RNA increases kinin B1 receptor expression and function in human airway epithelial cells

International Immunopharmacology ◽

10.1016/j.intimp.2007.07.002 ◽

2007 ◽

Vol 7 (14) ◽

pp. 1880-1887 ◽

Cited By ~ 5

Author(s):

S.H. Bengtson ◽

J. Eddleston ◽

S.C. Christiansen ◽

B.L. Zuraw

Keyword(s):

Epithelial Cells ◽

Airway Epithelial Cells ◽

Receptor Expression ◽

Airway Epithelial ◽

Double Stranded Rna ◽

Human Airway ◽

B1 Receptor ◽

Kinin B1 Receptor ◽

And Function ◽

Human Airway Epithelial Cells

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Arsenic alters tight junction expression, distribution and function in human airway epithelial cells

The FASEB Journal ◽

10.1096/fasebj.23.1_supplement.996.3 ◽

2009 ◽

Vol 23 (S1) ◽

Author(s):

Cara L. Sherwood ◽

Andrew E. Liguori ◽

Colin E. Olsen ◽

R. Clark Lantz ◽

Jefferey L. Burgess ◽

...

Keyword(s):

Epithelial Cells ◽

Tight Junction ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Human Airway ◽

And Function ◽

Human Airway Epithelial Cells

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Prolonged cigarette smoke exposure alters mitochondrial structure and function in airway epithelial cells

Respiratory Research ◽

10.1186/1465-9921-14-97 ◽

2013 ◽

Vol 14 (1) ◽

pp. 97 ◽

Cited By ~ 117

Author(s):

Roland F Hoffmann ◽

Sina Zarrintan ◽

Simone M Brandenburg ◽

Arjan Kol ◽

Harold G de Bruin ◽

...

Keyword(s):

Epithelial Cells ◽

Cigarette Smoke ◽

Airway Epithelial Cells ◽

Structure And Function ◽

Smoke Exposure ◽

Cigarette Smoke Exposure ◽

Airway Epithelial ◽

Mitochondrial Structure ◽

And Function

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Identification and function of A1 adenosine receptors in normal and cystic fibrosis human airway epithelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.1995.268.6.c1520 ◽

1995 ◽

Vol 268 (6) ◽

pp. C1520-C1527 ◽

Cited By ~ 22

Author(s):

D. E. McCoy ◽

E. M. Schwiebert ◽

K. H. Karlson ◽

W. S. Spielman ◽

B. A. Stanton

Keyword(s):

Cystic Fibrosis ◽

Epithelial Cells ◽

Patch Clamp ◽

Airway Epithelial Cells ◽

Airway Epithelial ◽

Whole Cell Patch Clamp ◽

Adenosine A1 ◽

A1 Receptor ◽

And Function ◽

Cl Conductance

A role for adenosine in the regulation of ion transport in pulmonary epithelial cells has recently been proposed. Although evidence exists documenting the presence and function of adenosine A2 receptors in airway epithelia, the presence of adenosine A1 receptors remains controversial. The present study used reverse transcriptase-polymerase chain reaction (PCR) and whole cell patch-clamp analysis to investigate A1 receptor presence and function in normal and cystic fibrosis (CF) human airway epithelial cells. Oligonucleotide primers complementary to the human brain A1 receptor sequence generated a PCR product of the predicted size (311 bp) in normal tracheal (9HTEo-) and CF submucosal (2CFSMEo-) airway cell lines and in primary cultures of CF nasal polyp epithelial cells. An oligonucleotide probe internal to the PCR primers hybridized with the 311-bp cDNAs by Southern blot analysis. cDNA sequencing demonstrated that the normal and CF airway cell PCR products are 100% identical to the corresponding sequence of the human brain adenosine A1 receptor. Northern blot analysis of 9HTEo-and 2CFSMEo- poly(A)+ RNA revealed the presence of two bands of approximately 3.0 and approximately 5.5 kb corresponding to the A1 receptor. Whole cell patch-clamp analyses demonstrated that 8-cyclopentyl-1,3-dipropylxanthine, a specific A1 receptor antagonist, increases adenosine 3',5'-cyclic monophosphate (cAMP)-activated Cl- conductance in 9HTEo-airway cells and allows cAMP to increase Cl- conductance in 2CFSMEo- CF airway cells and CF nasal polyp epithelial cells in primary culture. These results provide evidence for the presence and function of A1 receptors in normal and CF airway epithelial cells and provide support for a role of adenosine A1 receptors in modulating airway epithelial cell Cl- transport.

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