title: R990G polymorphism of calcium-sensing receptor does produce a gain-of-function and predispose to primary hypercalciuria.
abstract: An association between the R990G polymorphism of the CaSR gene, coding for calcium-sensing receptor, and primary hypercalciuria was found in kidney stone formers. To confirm this relationship, we investigated hypercalciuric women without stones and studied the effect of CaSR gene in human embryonic kidney cells (HEK-293). We genotyped for CaSR A986S, R990G, and Q1011E polymorphisms, 119 normocalciuric and 124 hypercalciuric women with negative history of kidney stones. Homozygous (n=2) or heterozygous (n=21) women for the 990G allele considered as one group had an increased risk to be hypercalciuric (odds ratio=5.2; P=0.001) and higher calcium excretion (P=0.005) in comparison with homozygous women for the 990R allele (n=220). HEK-293 cells were transfected with the variant allele at the three CaSR gene polymorphisms and with the most common allele with no variants. The transient increment of intracellular calcium caused by the stepwise increase of extracellular calcium was evaluated in stable transfected cells loaded with fura-2 AM. The extracellular calcium concentration producing the half-maximal intracellular calcium response was lower in HEK-293 cells transfected with the 990G allele than in those transfected with the wild-type allele (P=0.0001). Our findings indicate that R990G polymorphism results in a gain-of-function of the calcium-sensing receptor and increased susceptibility to primary hypercalciuria.
authors.affiliation: Nephrology and Dialysis Unit, Postgraduate School of Nephrology, Vita Salute University, IRCCS San Raffaele Hospital, Milan, Italy. vexxoli.giuseppe@hsr.it
authors.forename: G
authors.initials: G
authors.lastname: Vezzoli
authors.forename: A
authors.initials: A
authors.lastname: Terranegra
authors.forename: T
authors.initials: T
authors.lastname: Arcidiacono
authors.forename: R
authors.initials: R
authors.lastname: Biasion
authors.forename: D
authors.initials: D
authors.lastname: Coviello
authors.forename: M L
authors.initials: ML
authors.lastname: Syren
authors.forename: V
authors.initials: V
authors.lastname: Paloschi
authors.forename: S
authors.initials: S
authors.lastname: Giannini
authors.forename: G
authors.initials: G
authors.lastname: Mignogna
authors.forename: A
authors.initials: A
authors.lastname: Rubinacci
authors.forename: A
authors.initials: A
authors.lastname: Ferraretto
authors.forename: D
authors.initials: D
authors.lastname: Cusi
authors.forename: G
authors.initials: G
authors.lastname: Bianchi
authors.forename: L
authors.initials: L
authors.lastname: Soldati
mesh_terms: D000483:Alleles; D019943:Amino Acid Substitution; D015153:Blotting, Western; D016022:Case-Control Studies; D002460:Cell Line; D003062:Codon; D004591:Electrophoresis, Polyacrylamide Gel; D005091:Exons; D005260:Female; D005456:Fluorescent Dyes; D016257:Fura-2 / Q000031:analogs & derivatives; D005787:Gene Frequency; D020022:Genetic Predisposition to Disease; D005998:Glycine / Q000378:metabolism; D006239:Haplotypes; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D053565:Hypercalciuria / Q000235:genetics*; D015810:Linkage Disequilibrium; D008875:Middle Aged; D016017:Odds Ratio; D011110:Polymorphism, Genetic; D020641:Polymorphism, Single Nucleotide; D017433:Protein Structure, Secondary; D044169:Receptors, Calcium-Sensing / Q000235:genetics*; D012307:Risk Factors; D014162:Transfection
chemical_list: D003062:Codon; D005456:Fluorescent Dyes; D044169:Receptors, Calcium-Sensing; C049925:fura-2-am; D005998:Glycine; D016257:Fura-2