Transient Receptor Potential Vanilloid Type 4 Channel Expression in Chronic Rhinosinusitis

Bhargave, Geeta; Woodworth, Bradford A.; Xiong, Guoxiang; Wolfe, Steven G.; Antunes, Marcelo B.; Cohen, Noam A.

doi:10.2500/ajr.2008.22.3125

Cited by 45 publications

(43 citation statements)

References 43 publications

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“…Primary sinonasal epithelial cells from humans, mice, and pigs, as well as CFBE immortalized cell lines expressing either wild type or F508del CFTR were cultured at an air-liquid interface according to previously established protocols[5,13-18] and used to evaluate whether resveratrol dependent CFTR activation is conserved across species. Murine nasal septal epithelial (MNSE) cells and a recombinant CFTR-expressing HEK293 cell line (“D060”) were employed to investigate effects of resveratrol on open probability (Po) of single CFTR ion channels.…”

Section: Methodsmentioning

confidence: 99%

Resveratrol Enhances Airway Surface Liquid Depth in Sinonasal Epithelium by Increasing Cystic Fibrosis Transmembrane Conductance Regulator Open Probability

et al. 2013

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BackgroundChronic rhinosinusitis engenders enormous morbidity in the general population, and is often refractory to medical intervention. Compounds that augment mucociliary clearance in airway epithelia represent a novel treatment strategy for diseases of mucus stasis. A dominant fluid and electrolyte secretory pathway in the nasal airways is governed by the cystic fibrosis transmembrane conductance regulator (CFTR). The objectives of the present study were to test resveratrol, a strong potentiator of CFTR channel open probability, in preparation for a clinical trial of mucociliary activators in human sinus disease. MethodsPrimary sinonasal epithelial cells, immortalized bronchoepithelial cells (wild type and F508del CFTR), and HEK293 cells expressing exogenous human CFTR were investigated by Ussing chamber as well as patch clamp technique under non-phosphorylating conditions. Effects on airway surface liquid depth were measured using confocal laser scanning microscopy. Impact on CFTR gene expression was measured by quantitative reverse transcriptase polymerase chain reaction.ResultsResveratrol is a robust CFTR channel potentiator in numerous mammalian species. The compound also activated temperature corrected F508del CFTR and enhanced CFTR-dependent chloride secretion in human sinus epithelium ex vivo to an extent comparable to the recently approved CFTR potentiator, ivacaftor. Using inside out patches from apical membranes of murine cells, resveratrol stimulated an ~8 picosiemens chloride channel consistent with CFTR. This observation was confirmed in HEK293 cells expressing exogenous CFTR. Treatment of sinonasal epithelium resulted in a significant increase in airway surface liquid depth (in µm: 8.08+/-1.68 vs. 6.11+/-0.47,control,p<0.05). There was no increase CFTR mRNA. ConclusionResveratrol is a potent chloride secretagogue from the mucosal surface of sinonasal epithelium, and hydrates airway surface liquid by increasing CFTR channel open probability. The foundation for a clinical trial utilizing resveratrol as a therapeutic intervention to increase mucociliary transport and airway surface liquid hydration in sinus disease is strongly supported by these findings.

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Section: Methodsmentioning

confidence: 99%

Resveratrol Enhances Airway Surface Liquid Depth in Sinonasal Epithelium by Increasing Cystic Fibrosis Transmembrane Conductance Regulator Open Probability

et al. 2013

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“…MNSE and HSNE cells were harvested, grown on Costar 6.5-mm diameter permeable filter supports (Corning Life Sciences, Lowell, MA), and submerged in culture medium as previously described. 7–10 Media was removed from the monolayers on day 4 after the epithelium reached confluence, and cells fed via the basal chamber. Transgenic CFTR −/− MNSE cultures were also investigated.…”

Section: Methodsmentioning

confidence: 99%

Cystic fibrosis transmembrane conductance regulator activation by the solvent ethanol: implications for topical drug delivery

Cho

Skinner

Zhang

et al. 2015

Int Forum Allergy Rhinol

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Background Decreased CFTR-mediated chloride (Cl) secretion across mucosal surfaces contributes to the development of airway disease by depleting airway surface liquid, increasing mucus viscosity and adhesion, and consequently hindering mucociliary clearance. We serendipitously discovered during testing of drugs solubilized in low concentrations ethanol (0.25%, 43mM) that the control vehicle produced robust activation of CFTR-mediated Cl− transport. The objective of the current study is to investigate low concentrations ethanol for effects on Cl− secretion and ciliary beat frequency (CBF). Methods Wild type (WT) and transgenic CFTR−/− primary murine nasoseptal epithelial (MNSE) and WT and F508del/F508del human sinonasal epithelial (HSNE) cultures were subjected to transepithelial ion transport measurements using pharmacologic manipulation in Ussing chambers. CBF activation was also monitored. Murine nasal potential difference (NPD) was measured in vivo. Results Ussing chamber tracings revealed ethanol activated CFTR-mediated Cl transport in a dose-dependent fashion in WT MNSE (n=4, p<0.05) and HSNE (n=4, p<0.05). Ethanol also significantly increased CBF (fold-change) in WT MNSE cultures in a dose dependent fashion [PBS, 1.33+/−0.04; 0.25% Ethanol, 1.37+/−0.09; 0.5% Ethanol, 1.53+/−0.06 (p<0.05), 1% Ethanol, 1.62+/−0.1 (p<0.05)]. Lack of stimulation in CFTR−/− and F508del/F508del cultures indicated activity was dependent on the presence of intact functional CFTR. Ethanol perfusion (0.5%) resulted in a significant −3.5mV mean NPD polarization when compared to control solution (p<0.05). Conclusion The observation that brief exposure of ethanol stimulated Cl− secretion via CFTR-mediated pathways indicates possible use as topical aerosol delivered alone or in combination with other CFTR activators for diseases of dysfunctional MCC in CRS.

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“…The culture technique for propagation of MNSE and HSNE at an air‐liquid interface has been described in our prior studies 17–21. Human sinus epithelium was derived from tissue removed during skull base surgery from individuals without inflammatory disease.…”

Section: Methodsmentioning

confidence: 99%

Exposure to cigarette smoke condensate reduces calcium activated chloride channel transport in primary sinonasal epithelial cultures

et al. 2010

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Background The cystic fibrosis transmembrane conductance regulator (CFTR) serves as a predominant Cl− transport conduit in airway epithelium and is inhibited by cigarette smoke in vitro and in vivo. Activation of secondary Cl− transport pathways through calcium-activated Cl− channels (CaCC) has been postulated as a mechanism to bypass defects in CFTR-mediated transport. Because it is not known whether CaCC’s are also inhibited by tobacco exposure, the current study was designed to investigate the effect of cigarette smoke condensate (CSC) on CaCC transport. Study Design In vitro study Methods Well-characterized primary murine nasal septal epithelial (MNSE) and human sinonasal epithelial (HSNE) cultures were exposed to CSC in Ussing chambers. We monitored CaCC short-circuit current through stimulation of P2Y purinergic receptors with UTP or ATP and selective inhibition of the CFTR dependent secretory pathway. Characterization of CaCC current was also accomplished in primary airway cells derived from transgenic CFTR−/− (knockout) murine models. Results Change in CaCC-mediated current (ΔISC - representing transepithelial Ca-mediated Cl− secretion in µA/cm2) was significantly decreased in CSC-exposed wild type MNSE when compared to controls{(32.8 +/− 4.6 vs. 47.5 +/− 2.3; respectively) p < 0.02}. A similar effect was demonstrated in CFTR−/− MNSE cultures(33.4 +/− 2.8 vs. 38.6 +/− 2.0; p<0.05}. HSNE cultures also had a significant reduction in ISC (16.1 +/− 0.6 vs. 22.7 +/− 0; p=0.008). Conclusions CSC affects multiple pathways fundamental to airway ion transport, including both cAMP and calcium activated Cl− transport. Inhibition of Cl− transport may contribute to common diseases of the airways, such as chronic rhinosinusitis and chronic obstructive pulmonary disease.

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Transient Receptor Potential Vanilloid Type 4 Channel Expression in Chronic Rhinosinusitis

Cited by 45 publications

References 43 publications

Resveratrol Enhances Airway Surface Liquid Depth in Sinonasal Epithelium by Increasing Cystic Fibrosis Transmembrane Conductance Regulator Open Probability

Resveratrol Enhances Airway Surface Liquid Depth in Sinonasal Epithelium by Increasing Cystic Fibrosis Transmembrane Conductance Regulator Open Probability

Cystic fibrosis transmembrane conductance regulator activation by the solvent ethanol: implications for topical drug delivery

Exposure to cigarette smoke condensate reduces calcium activated chloride channel transport in primary sinonasal epithelial cultures

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