Reproducibility and intraindividual variation over days in buccal cell DNA methylation of two asthma genes, interferon γ (IFNγ) and inducible nitric oxide synthase (iNOS) 英文参考文献.docVIP

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Reproducibility and intraindividual variation over days in buccal cell DNA methylation of two asthma genes, interferon γ (IFNγ) and inducible nitric oxide synthase (iNOS) 英文参考文献.doc

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Reproducibility and intraindividual variation over days in buccal cell DNA methylation of two asthma genes, interferon γ (IFNγ) and inducible nitric oxide synthase (iNOS) 英文参考文献

Torroneetal.ClinicalEpigenetics2012,4:3 /content/4/1/3 RESEARCH OpenAccess Reproducibilityandintraindividualvariationover daysinbuccalcellDNAmethylationoftwo asthmagenes,interferong(IFNg)andinducible nitricoxidesynthase(iNOS) DZTorrone1,JSKuriakose1,KMoors1,HJiang1,MMNiedzwiecki2,FFPerera2andRLMiller1,2,3* Abstract Thebiologicalmechanismsresponsiblefortheonsetandexacerbationofasthmasymptomsinchildrenmay involvetheepigeneticregulationofinflammatorygenesafterenvironmentalexposures.Usingbuccalcells,we hypothesizedthatDNAmethylationinpromoterregionsoftwoasthmagenes,induciblenitricoxidesynthase (iNOS)andinterferong(IFNg),canvaryoverseveraldays.Repeatbuccalsampleswerecollected4to7daysapart from34childrenparticipatingintheColumbiaCenterforChildren’sEnvironmentalHealth(CCCEH)birthcohort study.Severalfieldduplicates(sequentialcollectionoftwosamplesinthefield)andreplicates(onesample pyrosequencedtwice)alsowerecollectedtoensureconsistencywithcollectionandlaboratoryprocedures.DNA methylationwasassessedbypyrosequencingaPCRofbisulfite-treatedDNA.Wefoundthatreplicateandfield duplicatesampleswerecorrelatedstrongly(r=0.86to0.99,P0.05),whilerepeatsamplesdemonstratedlow within-subjectcorrelations(r=0.19to0.56,P=0.06to0.30).OurdatarevealDNAmethylationasadynamic epigeneticmechanismthatcanbeaccessedsafelyandreproduciblyinaninnercitypediatriccohortusingnon- invasivebuccalswabsandpyrosequencingtechnology. Keywords:methylation,asthma,IFNγ,iNOS,buccalmucosa,epigeneticregulation,pediatric,innercity Introduction Thebiologicalmechanismsresponsibleforthedevelop- ment of asthma symptoms in children following acute altered asthma gene expression and asthma symptoms [10-14]. To date, clinical research on epigenetic changes in exposuretoairpollutionandothertriggersarecomplex. asthma and other complex diseases has been limited, Theseincludetheinductionofoxidativestresspathways especiallyinchildren[11,12].Onecross-sectionalstudy and formation of excessive reactive oxygen species in byWhit