PUBLICATION

A Remote Cis-Acting Variant at 3q Links Glomerular NCK1 to Diabetic Nephropathy

Authors
He, B., Osterholm, A.M., Ojala, J.R., Andersson, A.C., and Tryggvason, K.
ID
ZDB-PUB-130312-17
Date
2013
Source
PLoS One   8(2): e56414 (Journal)
Registered Authors
He, Bing, Tryggvason, Karlynn
Keywords
none
MeSH Terms
  • Adaptor Proteins, Signal Transducing/genetics*
  • Animals
  • Animals, Genetically Modified
  • Chromosomes, Human, Pair 3*
  • Diabetic Nephropathies/genetics*
  • Gene Expression
  • Gene Expression Regulation
  • Gene Order
  • Genes, Reporter
  • Genetic Variation*
  • Genotype
  • Humans
  • Kidney Glomerulus/metabolism*
  • Kidney Glomerulus/pathology
  • Mice
  • Oncogene Proteins/genetics*
  • Polymorphism, Single Nucleotide
  • Regulatory Sequences, Nucleic Acid*
  • Zebrafish
PubMed
23441190 Full text @ PLoS One
Abstract

We have previously reported genetic association of a single nucleotide polymorphism (SNP), rs1866813, at 3q locus with increased risk of diabetic nephropathy (DN). The SNP is located approximately 70 kb downstream of a cluster of four genes. This raises a question how the remote noncoding polymorphism affects the risk of DN. In this study, we tested a long-range regulatory potential of this variant by a series of experiments. In a luciferase assay, two alleles of the SNP showed differential effects on the luciferase activity in transfected cells in vitro. Using transgenic zebrafish, we further demonstrated in vivo that two alleles of the SNP differentially regulated GFP expression in zebrafish podocytes. Immunofluorescence staining and Western blotting verified that only Nck1 of the four nearby genes was predominantly expressed in mouse glomeruli as well as in podocytes. Furthermore, genotypes of the SNP rs1866813 were correlated with NCK1 expression in immortalized lymphocytes from diabetic patients. The risk allele was associated with increased NCK1 expression compared to the non-risk allele, consistent with the results of the reporter-based studies. Interestingly, differential expression of glomerular Nck1 between mouse strains carrying the nephropathy-prone 129/Sv allele and nephropathy-resistant C57BL/6 allele was also observed. Our results show that the DN-associated SNP rs1866813 is a remote cis-acting variant differentially regulating glomerular NCK1 expression. This finding implicates an important role for glomerular NCK1 in DN pathogenesis under hyperglycemia.

Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping