title: Expression of the osteoarthritis-associated gene GDF5 is modulated epigenetically by DNA methylation.
abstract: GDF5 is involved in synovial joint development, maintenance and repair, and the rs143383 C/T single nucleotide polymorphism (SNP) located in the 5'UTR of GDF5 is associated, at the genome-wide significance level, with osteoarthritis susceptibility, and with other musculoskeletal phenotypes including height, congenital hip dysplasia and Achilles tendinopathy. There is a significant reduction in the expression of the disease-associated T allele relative to the C allele in synovial joint tissues, an effect influenced by a second SNP (rs143384, C/T) also within the 5'UTR. The differential allelic expression (DAE) imbalance of the C and T alleles of rs143383 varies intra- and inter-individually, suggesting that DAE may be modulated epigenetically. The C alleles of both SNPs form CpG dinucleotides that are potentially amenable to regulation by methylation. Here, we have examined whether DNA methylation regulates GDF5 expression and the allelic imbalance caused by rs143383. We observed methylation of the GDF5 promoter and 5'UTR in cell lines and joint tissues, with demethylation correlating with increased GDF5 expression. The CpG sites created by the C alleles at rs143383 and rs143384 were variably methylated, and treatment of a heterozygous cell line with a demethylating agent further increased the allelic expression imbalance between the C and T alleles. This demonstrates that the genetic effect of the rs143383 SNP on GDF5 expression is modulated epigenetically by DNA methylation. The variability in DAE of rs143383 is therefore partly accounted for by differences in DNA methylation that could influence the penetrance of this allele in susceptibility to common musculoskeletal diseases.
authors.affiliation: Musculoskeletal Research Group, Institute of Cellular Medicine, Newcastle University, 4th Floor Catherine Cookson Building, The Medical School, Framlington Place, Newcastle upon Tyne NE2 4HH, UK. louise.reynard@ncl.ac.uk
authors.forename: Louise N
authors.initials: LN
authors.lastname: Reynard
authors.forename: Catherine
authors.initials: C
authors.lastname: Bui
authors.forename: Elizabeth G
authors.initials: EG
authors.lastname: Canty-Laird
authors.forename: David A
authors.initials: DA
authors.lastname: Young
authors.forename: John
authors.initials: J
authors.lastname: Loughlin
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics; D002460:Cell Line; D045744:Cell Line, Tumor; D019175:DNA Methylation / Q000235:genetics*; D020022:Genetic Predisposition to Disease / Q000235:genetics; D055428:Growth Differentiation Factor 5 / Q000235:genetics*; D006801:Humans; D010003:Osteoarthritis / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D020121:5' Untranslated Regions; D055428:Growth Differentiation Factor 5