Lin, JinHeng and Gettings, Sean M. and Talbi, Khaoula and Schreiber, Rainer and Taggart, Michael J. and Preller, Matthias and Kunzelmann, Karl and Althaus, Mike and Gray, Michael A. (2023) Pharmacological inhibitors of the cystic fibrosis transmembrane conductance regulator exert off-target effects on epithelial cation channels. PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 475 (2). pp. 167-179. ISSN 0031-6768, 1432-2013
Full text not available from this repository. (Request a copy)Abstract
The cystic fibrosis transmembrane conductance regulator (CFTR) anion channel and the epithelial Na+ channel (ENaC) play essential roles in transepithelial ion and fluid transport in numerous epithelial tissues. Inhibitors of both channels have been important tools for defining their physiological role in vitro. However, two commonly used CFTR inhibitors, CFTRinh-172 and GlyH-101, also inhibit non-CFTR anion channels, indicating they are not CFTR specific. However, the potential off-target effects of these inhibitors on epithelial cation channels has to date not been addressed. Here, we show that both CFTR blockers, at concentrations routinely employed by many researchers, caused a significant inhibition of store-operated calcium entry (SOCE) that was time-dependent, poorly reversible and independent of CFTR. Patch clamp experiments showed that both CFTRinh-172 and GlyH-101 caused a significant block of Orai1-mediated whole cell currents, establishing that they likely reduce SOCE via modulation of this Ca2+ release-activated Ca2+ (CRAC) channel. In addition to off-target effects on calcium channels, both inhibitors significantly reduced human alpha beta gamma-ENaC-mediated currents after heterologous expression in Xenopus oocytes, but had differential effects on delta beta gamma-ENaC function. Molecular docking identified two putative binding sites in the extracellular domain of ENaC for both CFTR blockers. Together, our results indicate that caution is needed when using these two CFTR inhibitors to dissect the role of CFTR, and potentially ENaC, in physiological processes.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CFTR INHIBITION; HCO3-SECRETION; ALPHA-SUBUNIT; CELL LINE; AMILORIDE; IDENTIFICATION; HOMEOSTASIS; CRACM1; CALU-3; CFTR inhibitors; Off-target effects; Store-operated calcium entry; Orai1; ENaC; In silico modelling |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Physiologie > Prof. Dr. Karl Kunzelmann |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 21 Feb 2024 07:04 |
| Last Modified: | 21 Feb 2024 07:04 |
| URI: | https://pred.uni-regensburg.de/id/eprint/58296 |
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