title: Tolvaptan inhibits ERK-dependent cell proliferation, Cl⁻ secretion, and in vitro cyst growth of human ADPKD cells stimulated by vasopressin.
abstract: In autosomal dominant polycystic kidney disease (ADPKD), arginine vasopressin (AVP) accelerates cyst growth by stimulating cAMP-dependent ERK activity and epithelial cell proliferation and by promoting Cl(-)-dependent fluid secretion. Tolvaptan, a V2 receptor antagonist, inhibits the renal effects of AVP and slows cyst growth in PKD animals. Here, we determined the effect of graded concentrations of tolvaptan on intracellular cAMP, ERK activity, cell proliferation, and transcellular Cl(-) secretion using human ADPKD cyst epithelial cells. Incubation of ADPKD cells with 10(-9) M AVP increased intracellular cAMP and stimulated ERK and cell proliferation. Tolvaptan caused a concentration-dependent inhibition of AVP-induced cAMP production with an apparent IC(50) of ∼10(-10) M. Correspondingly, tolvaptan inhibited AVP-induced ERK signaling and cell proliferation. Basolateral application of AVP to ADPKD cell monolayers grown on permeable supports caused a sustained increase in short-circuit current that was completely blocked by the Cl(-) channel blocker CFTR(inh-172), consistent with AVP-induced transepithelial Cl(-) secretion. Tolvaptan inhibited AVP-induced Cl(-) secretion and decreased in vitro cyst growth of ADPKD cells cultured within a three-dimensional collagen matrix. These data demonstrate that relatively low concentrations of tolvaptan inhibit AVP-stimulated cell proliferation and Cl(-)-dependent fluid secretion by human ADPKD cystic cells.
authors.affiliation: Department of Medicine, University of Kansas Medical Center, Kansas City, Kansas 66160-3018, USA.
authors.forename: Gail A
authors.initials: GA
authors.lastname: Reif
authors.forename: Tamio
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Emily
authors.initials: E
authors.lastname: Nivens
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Fujiki
authors.forename: Cibele S
authors.initials: CS
authors.lastname: Pinto
authors.forename: Darren P
authors.initials: DP
authors.lastname: Wallace
mesh_terms: D000328:Adult; D000368:Aged; D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology; D065092:Antidiuretic Hormone Receptor Antagonists; D001552:Benzazepines / Q000494:pharmacology*; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D002712:Chlorides / Q000378:metabolism*; D000242:Cyclic AMP / Q000096:biosynthesis; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism; D003560:Cysts / Q000473:pathology*; D004232:Diuretics / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D016891:Polycystic Kidney, Autosomal Dominant / Q000378:metabolism*; D012076:Renal Agents / Q000494:pharmacology*; D000077602:Tolvaptan; D014667:Vasopressins / Q000494:pharmacology*
chemical_list: D065092:Antidiuretic Hormone Receptor Antagonists; D001552:Benzazepines; D002712:Chlorides; D004232:Diuretics; D012076:Renal Agents; C013407:benzamil; D014667:Vasopressins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000077602:Tolvaptan; D000584:Amiloride; D000242:Cyclic AMP; D048049:Extracellular Signal-Regulated MAP Kinases