title: Detection of β cell death in diabetes using differentially methylated circulating DNA.
abstract: In diabetes mellitus, β cell destruction is largely silent and can be detected only after significant loss of insulin secretion capacity. We have developed a method for detecting β cell death in vivo by amplifying and measuring the proportion of insulin 1 DNA from β cells in the serum. By using primers that are specific for DNA methylation patterns in β cells, we have detected circulating copies of β cell-derived demethylated DNA in serum of mice by quantitative PCR. Accordingly, we have identified a relative increase of β cell-derived DNA after induction of diabetes with streptozotocin and during development of diabetes in nonobese diabetic mice. We have extended the use of this assay to measure β cell-derived insulin DNA in human tissues and serum. We found increased levels of demethylated insulin DNA in subjects with new-onset type 1 diabetes compared with age-matched control subjects. Our method provides a noninvasive approach for detecting β cell death in vivo that may be used to track the progression of diabetes and guide its treatment.
authors.affiliation: Department of Immunobiology and Internal Medicine, Yale University School of Medicine, New Haven, CT 06511, USA.
authors.forename: Eitan M
authors.initials: EM
authors.lastname: Akirav
authors.forename: Jasmin
authors.initials: J
authors.lastname: Lebastchi
authors.forename: Eva M
authors.initials: EM
authors.lastname: Galvan
authors.forename: Octavian
authors.initials: O
authors.lastname: Henegariu
authors.forename: Michael
authors.initials: M
authors.lastname: Akirav
authors.forename: Vitaly
authors.initials: V
authors.lastname: Ablamunits
authors.forename: Paul M
authors.initials: PM
authors.lastname: Lizardi
authors.forename: Kevan C
authors.initials: KC
authors.lastname: Herold
mesh_terms: D000818:Animals; D001483:Base Sequence; D016923:Cell Death / Q000502:physiology*; D003001:Cloning, Molecular; D004247:DNA / Q000097:blood* / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D003920:Diabetes Mellitus / Q000097:blood / Q000473:pathology*; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007328:Insulin / Q000235:genetics*; D050417:Insulin-Secreting Cells / Q000737:chemistry / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D016688:Mice, Inbred NOD; D008969:Molecular Sequence Data; D008991:Monitoring, Physiologic / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers; D007328:Insulin; D004247:DNA
references.citation: Cancer Res. 2003 Nov 15;63(22):7641-5
references.pmid: 14633683
references.citation: J Physiol. 1939 Nov 14;97(1):107-19
references.pmid: 16995144
references.citation: Curr Protoc Cytom. 2011 Jan;Chapter 6:6.27.1-6.27.16
references.pmid: 21207358
references.citation: Diabetes. 2006 Dec;55(12):3238-45
references.pmid: 17130466
references.citation: Diabetes. 1995 Mar;44(3):306-13
references.pmid: 7533732
references.citation: Clin Cancer Res. 2006 Dec 15;12(24):7347-52
references.pmid: 17189406
references.citation: Cell. 2010 Oct 29;143(3):470-84
references.pmid: 21029866
references.citation: Diabetes Care. 2008 Oct;31(10):1966-71
references.pmid: 18628574
references.citation: Diabetes Care. 1992 Aug;15(8):1009-13
references.pmid: 1505301
references.citation: Cancer Res. 2001 Feb 1;61(3):900-2
references.pmid: 11221878
references.citation: Magn Reson Med. 2008 Apr;59(4):712-20
references.pmid: 18302224
references.citation: N Engl J Med. 2005 Jun 23;352(25):2598-608
references.pmid: 15972866
references.citation: Acta Diabetol Lat. 1982 Oct-Dec;19(4):351-8
references.pmid: 6758459
references.citation: Am J Transplant. 2006 Jul;6(7):1704-11
references.pmid: 16827874
references.citation: Biotechniques. 2007 Jul;43(1 Suppl):25-30
references.pmid: 17936939
references.citation: N Engl J Med. 1988 Mar 17;318(11):663-70
references.pmid: 3125434
references.citation: J Cell Physiol. 2007 Nov;213(2):384-90
references.pmid: 17708532
references.citation: Diabetes. 1996 Jul;45(7):926-33
references.pmid: 8666144
references.citation: Diabetes. 2000 Jan;49(1):1-7
references.pmid: 10615942
references.citation: Nat Clin Pract Endocrinol Metab. 2008 Jun;4(6):334-43
references.pmid: 18446141
references.citation: Diabetes. 2008 Nov;57(11):2883-8
references.pmid: 18971435
references.citation: Am J Surg. 1988 Sep;156(3 Pt 1):191-3
references.pmid: 3048134
references.citation: Diabetes Obes Metab. 2007 Nov;9 Suppl 2:14-22
references.pmid: 17919174
references.citation: Trends Biochem Sci. 2006 Feb;31(2):89-97
references.pmid: 16403636
references.citation: Diabetes Care. 2007 Jan;30(1):38-42
references.pmid: 17192330
references.citation: Trends Mol Med. 2007 Sep;13(9):373-80
references.pmid: 17822959
references.citation: J Clin Invest. 2010 Nov;120(11):4031-9
references.pmid: 20978346
references.citation: J Clin Invest. 1995 Apr;95(4):1505-11
references.pmid: 7706455
references.citation: Nat Genet. 1994 Mar;6(3):310-3
references.pmid: 8012396
references.citation: Nature. 2010 Apr 29;464(7293):1293-300
references.pmid: 20432533
references.citation: PLoS One. 2009 Sep 09;4(9):e6953
references.pmid: 19742322
references.citation: Endocrinol Metab Clin North Am. 2009 Jun;38(2):303-17, viii
references.pmid: 19328413
references.citation: Diabetes. 2004 Feb;53(2):426-33
references.pmid: 14747294
references.citation: Diabetes. 2008 Apr;57(4):1084-92
references.pmid: 18252895