title: Novel activating mutations of the calcium-sensing receptor: the calcilytic NPS-2143 mitigates excessive signal transduction of mutant receptors.
abstract: OBJECTIVE
  Activating mutations in the calcium-sensing receptor (CaSR) gene cause autosomal dominant hypocalcemia (ADH). The aims of the present study were the functional characterization of novel mutations of the CaSR found in patients, the comparison of in vitro receptor function with clinical parameters, and the effect of the allosteric calcilytic NPS-2143 on the signaling of mutant receptors as a potential new treatment for ADH patients.
  
  
  METHODS
  Wild-type and mutant CaSR (T151R, P221L, E767Q, G830S, and A844T) were expressed in human embryonic kidney cells (HEK 293T). Receptor signaling was studied by measuring intracellular free calcium in response to different concentrations of extracellular calcium ([Ca(2+)](o)) in the presence or absence of NPS-2143.
  
  
  RESULTS
  All ADH patients had lowered serum calcium ranging from 1.7 to 2.0 mm and inadequate intact PTH and urinary calcium excretion. In vitro testing of CaSR mutations from these patients revealed exaggerated [Ca(2+)](o)-induced cytosolic Ca(2+) responses with EC(50) values for [Ca(2+)](o) ranging from 1.56 to 3.15 mM, which was lower than for the wild-type receptor (4.27 mM). The calcilytic NPS-2143 diminished the responsiveness to [Ca(2+)](o) in the CaSR mutants T151R, E767Q, G830S, and A844T. The mutant P221L, however, was only responsive when coexpressed with the wild-type CaSR.
  
  
  CONCLUSIONS
  Calcilytics might offer medical treatment for patients with autosomal dominant hypocalcemia caused by calcilytic-sensitive CaSR mutants.
authors.affiliation: Division of Endocrinology and Diabetes, Department of Medicine I, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Saskia
authors.initials: S
authors.lastname: Letz
authors.forename: Ramona
authors.initials: R
authors.lastname: Rus
authors.forename: Christine
authors.initials: C
authors.lastname: Haag
authors.forename: Helmuth-Günther
authors.initials: HG
authors.lastname: Dörr
authors.forename: Dirk
authors.initials: D
authors.lastname: Schnabel
authors.forename: Matthias
authors.initials: M
authors.lastname: Möhlig
authors.forename: Egbert
authors.initials: E
authors.lastname: Schulze
authors.forename: Karin
authors.initials: K
authors.lastname: Frank-Raue
authors.forename: Friedhelm
authors.initials: F
authors.lastname: Raue
authors.forename: Bernhard
authors.initials: B
authors.lastname: Mayr
authors.forename: Christof
authors.initials: C
authors.lastname: Schöfl
mesh_terms: D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D006801:Humans; D006996:Hypocalcemia / Q000235:genetics; D009154:Mutation / Q000502:physiology; D009281:Naphthalenes / Q000494:pharmacology*; D044169:Receptors, Calcium-Sensing / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D014162:Transfection
chemical_list: C436740:N-(2-hydroxy-3-(2-cyano-3-chlorophenoxy)propyl)-1,1-dimethyl-2-(2-nephthyl)ethylamine; D009281:Naphthalenes; D044169:Receptors, Calcium-Sensing; D002118:Calcium