Age-related loss of CpG methylation in the tumour necrosis factor promoter

Background: Dysregulated production of TNF has been implicated in the pathogenesis and severity of inflammatory rheumatic diseases, many of which show age-related increased incidence. Ageing is also associated with changes in the immune system including higher systemic levels of pro-inflammatory cytokines. Methylation of DNA is an important regulator of gene expression and changes with age.Objective: In this study we investigated whether the DNA methylation status of the TNF promoter changed with age in peripheral blood leucocytes and macrophages.Methods and results: Using pyrosequencing assays we detected age-related demethylation of CpG motifs (−304, −245 and −239) in the TNF promoter in human peripheral blood cells from 312 healthy controls (0.8% per decade, confidence interval (CI) = 0.44-1.13%, p = 1 × 10−5) and primary monocyte-derived macrophages (MDM) from a separate population of 78 healthy controls (1.4% per decade, CI = 0.79-2.13%, p = 7 × 10−5). Methylation a TNF promoter fragment (−345-+154) resulted in 78% reduction of reporter gene activity compared with the unmethylated promoter construct.Conclusions: These data suggest a potential role of accrued changes in DNA methylation in the development of age-related inflammatory diseases, such as rheumatoid arthritis and polymyalgia rheumatica, in which TNF is a pivotal mediator.

► Demethylation of TNF 5′ region occurs with ageing in peripheral blood leucocytes and macrophages. ► DNA demethylation of HeLA cells leads to increased LPS-induced TNF mRNA levels. ► Demethylation of the 5′ region leads to increased activity of a TNF promoter reporter gene construct.