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However, few epigenetic modifications directly implicated in such processes have been identified during adolescence, a critical developmental window where physiological changes could influence future disease trajectory. To investigate the association between DNA methylation and NAFLD in adolescence, we undertook discovery and validation of novel methylation marks, alongside replication of previously reported marks.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Approach and results<\/jats:title>\n                    <jats:p>\n                      We performed a DNA methylation epigenome-wide association study (EWAS) on DNA from whole blood from 707 Raine Study adolescents phenotyped for steatosis score and NAFLD by ultrasound at age 17. Next, we performed pyrosequencing validation of loci within the most 100 strongly associated differentially methylated CpG sites (dmCpGs) for which\u2009\u2265\u20092 probes per gene remained significant across four statistical models with a nominal\n                      <jats:italic>p<\/jats:italic>\n                      value\u2009&lt;\u20090.007. EWAS identified dmCpGs related to three genes (\n                      <jats:italic>ANK1, MIR10a<\/jats:italic>\n                      ,\n                      <jats:italic>PTPRN2<\/jats:italic>\n                      ) that met our criteria for pyrosequencing. Of the dmCpGs and surrounding loci that were pyrosequenced (\n                      <jats:italic>ANK1 n<\/jats:italic>\n                      \u2009=\u20096,\n                      <jats:italic>MIR10a n<\/jats:italic>\n                      \u2009=\u20097,\n                      <jats:italic>PTPRN2 n<\/jats:italic>\n                      \u2009=\u20093), three dmCpGs in\n                      <jats:italic>ANK1<\/jats:italic>\n                      and two in\n                      <jats:italic>MIR10a<\/jats:italic>\n                      were significantly associated with NAFLD in adolescence. After adjustment for waist circumference only dmCpGs in\n                      <jats:italic>ANK1<\/jats:italic>\n                      remained significant. These\n                      <jats:italic>ANK1<\/jats:italic>\n                      CpGs were also associated with \u03b3-glutamyl transferase and alanine aminotransferase concentrations. Three of twenty-two differentially methylated dmCpGs previously associated with adult NAFLD were associated with NAFLD in adolescence (all adjusted\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&lt;\u20092.3\u2009\u00d7\u200910\n                      <jats:sup>\u20133<\/jats:sup>\n                      ).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>We identified novel DNA methylation loci associated with NAFLD and serum liver biochemistry markers during adolescence, implicating putative dmCpG\/gene regulatory pathways and providing insights for future mechanistic studies.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1007\/s12072-022-10469-7","type":"journal-article","created":{"date-parts":[[2023,2,3]],"date-time":"2023-02-03T14:02:31Z","timestamp":1675432951000},"page":"584-594","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Differential DNA methylation of steatosis and non-alcoholic fatty liver disease in adolescence"],"prefix":"10.1007","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4026-2964","authenticated-orcid":false,"given":"Phillip E.","family":"Melton","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. 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G.C.B. has received research funding from Nestle, Abbott Nutrition and Danone. He has served as a member of the Scientific Advisory Board of BASF and is a member of the BASF Asia\u2013Pacific Grant Award Panel.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}},{"value":"The study was approved by the University of Western Australia Human Ethics Committee and all participants gave their written informed consent.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Informed consent"}}]}}