Decreased transcriptional activity of calcium-sensing receptor gene promoter 1 is associated with calcium nephrolithiasis

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Abstract

Background: CaSR gene is a candidate for calcium nephrolithiasis. Single-nucleotide polymorphisms (SNPs) encompassing its regulatory region were associated with calcium nephrolithiasis. Aims: We tested SNPs in the CaSR gene regulatory region associated with calcium nephrolithiasis and their effects in kidney. Subjects and Methods: One hundred sixty-seven idiopathic calcium stone formers and 214 healthy controls were genotyped for four CaSR gene SNPs identified by bioinformatics analysis as modifying transcription factor binding sites. Strontium excretion afteranoral loadwastested in 55 stone formers. Transcriptional activity induced by variant alleles at CaSR gene promoters was compared by luciferase reporter gene assay in HEK-293 and HKC-8 cells. CaSR and claudin-14 mRNA levels were measured by real-Time PCR in 107 normal kidney medulla samples and compared in patients with different CaSR genotype. Results: Only rs6776158 (A>G), located in the promoter 1, was associated with nephrolithiasis. Its minor G allele was more frequent in stone formers than controls (37.8% vs 26.4%, P = .001). A reduced strontium excretion was observed in GG homozygous stone formers. Luciferase fluorescent activity was lower in cells transfected with the promoter 1 including G allele at rs6776158 than cells transfected with the A allele. CaSRmRNAlevels were lower in kidney medulla samples from homozygous carriers for theGallele at rs6776158 than carriers for the A allele. Claudin-14 mRNA levels were also lower in GG homozygous subjects. Conclusions: Minor allele at rs6776158 may predispose to calcium stones by decreasing transcriptional activity of theCaSRgenepromoter 1andCaSRexpression in kidney tubules.

Original languageEnglish
Pages (from-to)3839-3847
Number of pages9
JournalJournal of Clinical Endocrinology and Metabolism
Volume98
Issue number9
DOIs
Publication statusPublished - 2013
Externally publishedYes

Fingerprint

Calcium-Sensing Receptors
Nephrolithiasis
Genes
Alleles
Calcium
Polymorphism
Strontium
Kidney Medulla
Nucleotides
Nucleic Acid Regulatory Sequences
Single Nucleotide Polymorphism
Luciferases
Messenger RNA
Kidney Tubules
Bioinformatics
Assays
Computational Biology
Reporter Genes
Transcription Factors
Binding Sites

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Endocrinology
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

@article{936f98c423364184ac610538501a8e85,
title = "Decreased transcriptional activity of calcium-sensing receptor gene promoter 1 is associated with calcium nephrolithiasis",
abstract = "Background: CaSR gene is a candidate for calcium nephrolithiasis. Single-nucleotide polymorphisms (SNPs) encompassing its regulatory region were associated with calcium nephrolithiasis. Aims: We tested SNPs in the CaSR gene regulatory region associated with calcium nephrolithiasis and their effects in kidney. Subjects and Methods: One hundred sixty-seven idiopathic calcium stone formers and 214 healthy controls were genotyped for four CaSR gene SNPs identified by bioinformatics analysis as modifying transcription factor binding sites. Strontium excretion afteranoral loadwastested in 55 stone formers. Transcriptional activity induced by variant alleles at CaSR gene promoters was compared by luciferase reporter gene assay in HEK-293 and HKC-8 cells. CaSR and claudin-14 mRNA levels were measured by real-Time PCR in 107 normal kidney medulla samples and compared in patients with different CaSR genotype. Results: Only rs6776158 (A>G), located in the promoter 1, was associated with nephrolithiasis. Its minor G allele was more frequent in stone formers than controls (37.8{\%} vs 26.4{\%}, P = .001). A reduced strontium excretion was observed in GG homozygous stone formers. Luciferase fluorescent activity was lower in cells transfected with the promoter 1 including G allele at rs6776158 than cells transfected with the A allele. CaSRmRNAlevels were lower in kidney medulla samples from homozygous carriers for theGallele at rs6776158 than carriers for the A allele. Claudin-14 mRNA levels were also lower in GG homozygous subjects. Conclusions: Minor allele at rs6776158 may predispose to calcium stones by decreasing transcriptional activity of theCaSRgenepromoter 1andCaSRexpression in kidney tubules.",
author = "{GENIAL network} and Giuseppe Vezzoli and Annalisa Terranegra and Andrea Aloia and Teresa Arcidiacono and Luciano Milanesi and Ettore Mosca and Alessandra Mingione and Donatella Spotti and Daniele Cusi and Jianghui Hou and Hendy, {Geoffrey N.} and Laura Soldati",
year = "2013",
doi = "10.1210/jc.2013-1834",
language = "English",
volume = "98",
pages = "3839--3847",
journal = "Journal of Clinical Endocrinology and Metabolism",
issn = "0021-972X",
publisher = "The Endocrine Society",
number = "9",

}

TY - JOUR

T1 - Decreased transcriptional activity of calcium-sensing receptor gene promoter 1 is associated with calcium nephrolithiasis

AU - GENIAL network

AU - Vezzoli, Giuseppe

AU - Terranegra, Annalisa

AU - Aloia, Andrea

AU - Arcidiacono, Teresa

AU - Milanesi, Luciano

AU - Mosca, Ettore

AU - Mingione, Alessandra

AU - Spotti, Donatella

AU - Cusi, Daniele

AU - Hou, Jianghui

AU - Hendy, Geoffrey N.

AU - Soldati, Laura

PY - 2013

Y1 - 2013

N2 - Background: CaSR gene is a candidate for calcium nephrolithiasis. Single-nucleotide polymorphisms (SNPs) encompassing its regulatory region were associated with calcium nephrolithiasis. Aims: We tested SNPs in the CaSR gene regulatory region associated with calcium nephrolithiasis and their effects in kidney. Subjects and Methods: One hundred sixty-seven idiopathic calcium stone formers and 214 healthy controls were genotyped for four CaSR gene SNPs identified by bioinformatics analysis as modifying transcription factor binding sites. Strontium excretion afteranoral loadwastested in 55 stone formers. Transcriptional activity induced by variant alleles at CaSR gene promoters was compared by luciferase reporter gene assay in HEK-293 and HKC-8 cells. CaSR and claudin-14 mRNA levels were measured by real-Time PCR in 107 normal kidney medulla samples and compared in patients with different CaSR genotype. Results: Only rs6776158 (A>G), located in the promoter 1, was associated with nephrolithiasis. Its minor G allele was more frequent in stone formers than controls (37.8% vs 26.4%, P = .001). A reduced strontium excretion was observed in GG homozygous stone formers. Luciferase fluorescent activity was lower in cells transfected with the promoter 1 including G allele at rs6776158 than cells transfected with the A allele. CaSRmRNAlevels were lower in kidney medulla samples from homozygous carriers for theGallele at rs6776158 than carriers for the A allele. Claudin-14 mRNA levels were also lower in GG homozygous subjects. Conclusions: Minor allele at rs6776158 may predispose to calcium stones by decreasing transcriptional activity of theCaSRgenepromoter 1andCaSRexpression in kidney tubules.

AB - Background: CaSR gene is a candidate for calcium nephrolithiasis. Single-nucleotide polymorphisms (SNPs) encompassing its regulatory region were associated with calcium nephrolithiasis. Aims: We tested SNPs in the CaSR gene regulatory region associated with calcium nephrolithiasis and their effects in kidney. Subjects and Methods: One hundred sixty-seven idiopathic calcium stone formers and 214 healthy controls were genotyped for four CaSR gene SNPs identified by bioinformatics analysis as modifying transcription factor binding sites. Strontium excretion afteranoral loadwastested in 55 stone formers. Transcriptional activity induced by variant alleles at CaSR gene promoters was compared by luciferase reporter gene assay in HEK-293 and HKC-8 cells. CaSR and claudin-14 mRNA levels were measured by real-Time PCR in 107 normal kidney medulla samples and compared in patients with different CaSR genotype. Results: Only rs6776158 (A>G), located in the promoter 1, was associated with nephrolithiasis. Its minor G allele was more frequent in stone formers than controls (37.8% vs 26.4%, P = .001). A reduced strontium excretion was observed in GG homozygous stone formers. Luciferase fluorescent activity was lower in cells transfected with the promoter 1 including G allele at rs6776158 than cells transfected with the A allele. CaSRmRNAlevels were lower in kidney medulla samples from homozygous carriers for theGallele at rs6776158 than carriers for the A allele. Claudin-14 mRNA levels were also lower in GG homozygous subjects. Conclusions: Minor allele at rs6776158 may predispose to calcium stones by decreasing transcriptional activity of theCaSRgenepromoter 1andCaSRexpression in kidney tubules.

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U2 - 10.1210/jc.2013-1834

DO - 10.1210/jc.2013-1834

M3 - Article

VL - 98

SP - 3839

EP - 3847

JO - Journal of Clinical Endocrinology and Metabolism

JF - Journal of Clinical Endocrinology and Metabolism

SN - 0021-972X

IS - 9

ER -