reproducibility and intraindividual variation over days in buccal cell dna methylation of two asthma genes, interferon γ (ifnγ) and inducible nitric oxide synthase (inos)再现性和个体内的变异在颊细胞dna甲基化两天哮喘基因,干扰素γ(ifnγ)和诱导一氧化氮合酶(间接宾语).pdfVIP

Torrone et al. Clinical Epigenetics 2012, 4:3 /content/4/1/3 RESEARCH Open Access Reproducibility and intraindividual variation over days in buccal cell DNA methylation of two asthma genes, interferon g (IFNg) and inducible nitric oxide synthase (iNOS) 1 1 1 1 2 2 1,2,3* DZ Torrone , JS Kuriakose , K Moors , H Jiang , MM Niedzwiecki , FF Perera and RL Miller Abstract The biological mechanisms responsible for the onset and exacerbation of asthma symptoms in children may involve the epigenetic regulation of inflammatory genes after environmental exposures. Using buccal cells, we hypothesized that DNA methylation in promoter regions of two asthma genes, inducible nitric oxide synthase (iNOS) and interferon g (IFNg), can vary over several days. Repeat buccal samples were collected 4 to 7 days apart from 34 children participating in the Columbia Center for Children’s Environmental Health (CCCEH) birth cohort study. Several field duplicates (sequential collection of two samples in the field) and replicates (one sample pyrosequenced twice) also were collected to ensure consistency with collection and laboratory procedures. DNA methylation was assessed by pyrosequencing a PCR of bisulfite-treated DNA. We found that replicate and field duplicate samples were correlated strongly (r = 0.86 to 0.99, P 0.05), while repeat samples demonstrated low within-subject correlations (r = 0.19 to 0.56, P = 0.06 to 0.30). Our data reveal DNA methylation as a dynamic epigenetic mechanism that can be accessed safely and reproducibly in an inner city pediatric cohort using non- invasive buccal swabs and pyrosequencing technology. Keywords: methylation, asthma, IFNγ, iNOS, buccal mucosa, epigenetic regulation, pediat