The distribution of two common TaqIrestriction fragment length polymorphisms (RFLPs) of the cholesteryl ester transfer protein (CETP) gene were determined in 271 Italian-born and 170 Greek-born migrants to Melbourne, Australia. A much smaller number were examined for the EcoNl RFLP of the same gene. Allele frequencies of the TaqlA RFLP exhibited the least variation in both ethnic groups, and no significant regional heterogeneity in allele or genotype frequencies of either TaqlRFLP was detected for Greece or Italy. There was no difference between Italians and Greeks for the Taql A polymorphism and the variability at the B RFLP was of borderline significance. Comparisons with other Caucasian populations revealed that allele frequencies of all three CETP RFLPs are remarkably uniform within Caucasians, with the TaqlB polymorphism being the most variable.

Objective. Osteoarthritis (OA) is polygenic, with more than 90 risk loci currently mapped, including at the singlenucleotide polymorphism rs6516886. Previous analysis of OA cartilage DNA identified 6 CpG dinucleotides with methylation levels that correlated with the rs6516886 genotype, forming methylation quantitative trait loci (mQTLs). We undertook this study to investigate these mQTLs and to map expression quantitative trait loci (eQTLs) across joint tissues in order to identify a particular gene as a target of the rs6516886 association effect. Methods. Nucleic acids were extracted from the cartilage, fat pad, synovium, and peripheral blood from OA patients. Methylation of CpGs and allelic expression imbalance of potential target genes were assessed by pyrosequencing. A chondrocyte cell line expressing deactivated Cas9 (dCas9)-TET1 was used to directly alter CpG methylation levels, with effects on gene expression quantified by polymerase chain reaction. Results. Multiple mQTLs were detected, with effects strongest in joint tissues and methylation at CpG cg20220242 correlating most significantly with the rs6516886 genotype. CpG cg20220242 is located upstream of RWDD2B. Significant rs6516886 eQTLs were observed for this gene, with the OA risk-conferring allele of rs6516886 correlating with reduced expression. CpG methylation also correlated with allelic expression of RWDD2B, forming methylation-expression QTLs (meQTLs). Deactivated Cas9-TET1 reduction in the methylation of cg20220242 increased expression of RWDD2B. Conclusion. The rs6516886 association signal is a multi-tissue meQTL involving cg20220242 and acting on RWDD2B. Modulating CpG methylation reverses the impact of the risk allele. RWDD2B codes for a protein about which little is currently known. Further analysis of RWDD2B as a target of OA genetic risk will provide novel insight into this complex disease.

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