{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T03:53:18Z","timestamp":1787025198295,"version":"build-2736575974"},"reference-count":16,"publisher":"American Diabetes Association","issue":"11","license":[{"start":{"date-parts":[[2016,5,1]],"date-time":"2016-05-01T00:00:00Z","timestamp":1462060800000},"content-version":"vor","delay-in-days":279,"URL":"http:\/\/www.diabetesjournals.org\/site\/license"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000901","name":"JDRF","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000901","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000041","name":"American Diabetes Association","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000041","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000989","name":"Ball Brothers Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000989","id-type":"DOI","asserted-by":"publisher"}]},{"name":"George and Francis Ball Foundation"}],"content-domain":{"domain":["diabetesjournals.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2015,11,1]]},"abstract":"<jats:p>Elevated ratios of circulating unmethylated to methylated preproinsulin (INS) DNA have been suggested to reflect \u03b2-cell death in type 1 diabetes (T1D). We tested the hypothesis that absolute levels (rather than ratios) of unmethylated and methylated INS DNA differ between subjects with new-onset T1D and control subjects and assessed longitudinal changes in these parameters. We used droplet digital PCR to measure levels of unmethylated and methylated INS DNA in serum from subjects at T1D onset and at 8 weeks and 1 year post-onset. Compared with control subjects, levels of both unmethylated and methylated INS DNA were elevated at T1D onset. At 8 weeks post-onset, methylated INS DNA remained elevated, but unmethylated INS DNA fell. At 1 year postonset, both unmethylated and methylated INS DNA returned to control levels. Subjects with obesity, type 2 diabetes, and autoimmune hepatitis exhibited lower levels of unmethylated and methylated INS compared with subjects with T1D at onset and no differences compared with control subjects. Our study shows that elevations in both unmethylated and methylated INS DNA occurs in new-onset T1D and that levels of these DNA species change during T1D evolution. Our work emphasizes the need to consider absolute levels of differentially methylated DNA species as potential biomarkers of disease.<\/jats:p>","DOI":"10.2337\/db15-0430","type":"journal-article","created":{"date-parts":[[2015,7,27]],"date-time":"2015-07-27T21:31:16Z","timestamp":1438032676000},"page":"3867-3872","update-policy":"https:\/\/doi.org\/10.2337\/ada-journal-policies","source":"Crossref","is-referenced-by-count":79,"title":["Elevations in Circulating Methylated and Unmethylated Preproinsulin DNA in New-Onset Type 1 Diabetes"],"prefix":"10.2337","volume":"64","author":[{"given":"Marisa M.","family":"Fisher","sequence":"first","affiliation":[{"name":"Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renecia A.","family":"Watkins","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janice","family":"Blum","sequence":"additional","affiliation":[{"name":"Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carmella","family":"Evans-Molina","sequence":"additional","affiliation":[{"name":"Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Medicine, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naga","family":"Chalasani","sequence":"additional","affiliation":[{"name":"Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Medicine, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, IN"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linda A.","family":"DiMeglio","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kieren J.","family":"Mather","sequence":"additional","affiliation":[{"name":"Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Medicine, Indiana University School of Medicine, Indianapolis, IN"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah A.","family":"Tersey","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raghavendra G.","family":"Mirmira","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Medicine, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, IN"},{"name":"Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1167","published-online":{"date-parts":[[2015,7,27]]},"reference":[{"key":"2022031303585610000_B1","doi-asserted-by":"crossref","first-page":"2883","DOI":"10.2337\/db07-1817","article-title":"Beta-cell mass and type 1 diabetes: going, going, gone","volume":"57","author":"Akirav","year":"2008","journal-title":"Diabetes"},{"key":"2022031303585610000_B2","doi-asserted-by":"crossref","first-page":"679","DOI":"10.2337\/db09-1378","article-title":"Progression to diabetes in relatives of type 1 diabetic patients: mechanisms and mode of onset","volume":"59","author":"Ferrannini","year":"2010","journal-title":"Diabetes"},{"key":"2022031303585610000_B3","doi-asserted-by":"crossref","first-page":"1173","DOI":"10.1586\/1744666X.2013.856757","article-title":"Predictors of progression to Type 1 diabetes: preparing for immune interventions in the preclinical disease phase","volume":"9","author":"Gorus","year":"2013","journal-title":"Expert Rev Clin Immunol"},{"key":"2022031303585610000_B4","doi-asserted-by":"crossref","first-page":"3476","DOI":"10.1210\/en.2013-1223","article-title":"Detection of islet \u03b2-cell death in vivo by multiplex PCR analysis of differentially methylated DNA","volume":"154","author":"Fisher","year":"2013","journal-title":"Endocrinology"},{"key":"2022031303585610000_B5","doi-asserted-by":"crossref","first-page":"19018","DOI":"10.1073\/pnas.1111008108","article-title":"Detection of \u03b2 cell death in diabetes using differentially methylated circulating DNA","volume":"108","author":"Akirav","year":"2011","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2022031303585610000_B6","doi-asserted-by":"crossref","first-page":"e94591","DOI":"10.1371\/journal.pone.0094591","article-title":"Tissue-specific methylation of human insulin gene and PCR assay for monitoring beta cell death","volume":"9","author":"Husseiny","year":"2014","journal-title":"PLoS One"},{"key":"2022031303585610000_B7","doi-asserted-by":"crossref","first-page":"e47942","DOI":"10.1371\/journal.pone.0047942","article-title":"Development of a quantitative methylation-specific polymerase chain reaction method for monitoring beta cell death in type 1 diabetes","volume":"7","author":"Husseiny","year":"2012","journal-title":"PLoS One"},{"key":"2022031303585610000_B8","doi-asserted-by":"crossref","first-page":"1676","DOI":"10.2337\/db12-1207","article-title":"Immune therapy and \u03b2-cell death in type 1 diabetes","volume":"62","author":"Lebastchi","year":"2013","journal-title":"Diabetes"},{"key":"2022031303585610000_B9","doi-asserted-by":"crossref","first-page":"3694","DOI":"10.1210\/en.2014-1150","article-title":"Analysis of \u03b2-cell death in type 1 diabetes by droplet digital PCR","volume":"155","author":"Usmani-Brown","year":"2014","journal-title":"Endocrinology"},{"key":"2022031303585610000_B10","doi-asserted-by":"crossref","first-page":"1163","DOI":"10.1172\/JCI78142","article-title":"\u03b2 cell death and dysfunction during type 1 diabetes development in at-risk individuals","volume":"125","author":"Herold","year":"2015","journal-title":"J Clin Invest"},{"key":"2022031303585610000_B11","doi-asserted-by":"crossref","first-page":"e6953","DOI":"10.1371\/journal.pone.0006953","article-title":"Insulin gene expression is regulated by DNA methylation","volume":"4","author":"Kuroda","year":"2009","journal-title":"PLoS One"},{"key":"2022031303585610000_B12","doi-asserted-by":"crossref","first-page":"360","DOI":"10.1007\/s00125-010-1967-6","article-title":"Insulin promoter DNA methylation correlates negatively with insulin gene expression and positively with HbA(1c) levels in human pancreatic islets","volume":"54","author":"Yang","year":"2011","journal-title":"Diabetologia"},{"key":"2022031303585610000_B13","doi-asserted-by":"crossref","first-page":"e36278","DOI":"10.1371\/journal.pone.0036278","article-title":"Association of the CpG methylation pattern of the proximal insulin gene promoter with type 1 diabetes","volume":"7","author":"Fradin","year":"2012","journal-title":"PLoS One"},{"key":"2022031303585610000_B14","doi-asserted-by":"crossref","first-page":"8604","DOI":"10.1021\/ac202028g","article-title":"High-throughput droplet digital PCR system for absolute quantitation of DNA copy number","volume":"83","author":"Hindson","year":"2011","journal-title":"Anal Chem"},{"key":"2022031303585610000_B15","doi-asserted-by":"crossref","first-page":"E866","DOI":"10.1210\/jc.2013-2221","article-title":"Syntaxin 4 up-regulation increases efficiency of insulin release in pancreatic islets from humans with and without type 2 diabetes mellitus","volume":"99","author":"Oh","year":"2014","journal-title":"J Clin Endocrinol Metab"},{"key":"2022031303585610000_B16","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1258\/acb.2010.009173","article-title":"Usefulness of cell-free plasma DNA, procalcitonin and C-reactive protein as markers of infection in febrile patients","volume":"47","author":"Moreira","year":"2010","journal-title":"Ann Clin Biochem"}],"container-title":["Diabetes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.org\/diabetes\/diabetes\/article-pdf\/64\/11\/3867\/576862\/db150430.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/diabetesjournals.org\/diabetes\/article-pdf\/64\/11\/3867\/576862\/db150430.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,11,2]],"date-time":"2022-11-02T12:46:56Z","timestamp":1667393216000},"score":1,"resource":{"primary":{"URL":"https:\/\/diabetesjournals.org\/diabetes\/article\/64\/11\/3867\/40294\/Elevations-in-Circulating-Methylated-and"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,7,27]]},"references-count":16,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2015,7,27]]},"published-print":{"date-parts":[[2015,11,1]]}},"URL":"https:\/\/doi.org\/10.2337\/db15-0430","relation":{"has-review":[{"id-type":"doi","id":"10.3410\/f.725678991.793513548","asserted-by":"object"}]},"ISSN":["0012-1797","1939-327X"],"issn-type":[{"value":"0012-1797","type":"print"},{"value":"1939-327X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,7,27]]}}}