No Evidence for Direct Activation of the Cystic Fibrosis Transmembrane Conductance Regulator by 8-Cyclopentyl-1,3-Dipropylxanthine

Abstract

8-Cyclopentyl-1,3-dipropylxanthine (CPX) is a selective A₁-adenosine receptor antagonist which has been reported to activate Cl^– efflux at very low concentrations in cells carrying the cystic fibrosis (CF) defect, but not in cells expressing the wild-type form of the CF transmembrane conductance regulator (CFTR). CPX was suggested as a new therapeutic drug for the treatment of CF. In the present study, we examined the effects of CPX on various types of recombinant cells (Xenopus oocytes, Chinese hamster ovary cells, CF tracheal cells) and native non-CF and CF respiratory epithelial cells. CPX did not activate a whole-cell conductance when applied at concentrations ranging from 1 nmol/l to 100 µmol/l in oocytes injected with water or expressing either wild-type CFTR or mutant ΔF508-CFTR. Correspondingly, CPX (10 µmol/l) did not activate whole-cell conductance in non-CF or CF respiratory epithelial cells and Chinese hamster ovary cells expressing either wild-type CFTR or ΔF508-CFTR. Instead, CPX depolarized V_m by inhibition of a K⁺ conductance in CF respiratory epithelial cells. At 10 µmol/l CPX marginally decreased intracellular pH in respiratory epithelial cells, independent of expression of wild-type CFTR or mutant CFTR. According to these data, CPX-induced ³⁶Cl efflux reported in previous studies cannot be attributed to direct activation of ΔF508-CFTR Cl^– conductance and is probably related to the CPX-induced changes in intracellular pH.