PMID- 30084829 OWN - NLM STAT- MEDLINE DCOM- 20180904 LR - 20211204 IS - 1939-327X (Electronic) IS - 0012-1797 (Print) IS - 0012-1797 (Linking) VI - 67 IP - 9 DP - 2018 Sep TI - The Dysregulation of the DLK1-MEG3 Locus in Islets From Patients With Type 2 Diabetes Is Mimicked by Targeted Epimutation of Its Promoter With TALE-DNMT Constructs. PG - 1807-1815 LID - 10.2337/db17-0682 [doi] AB - Type 2 diabetes mellitus (T2DM) is characterized by the inability of the insulin-producing beta-cells to overcome insulin resistance. We previously identified an imprinted region on chromosome 14, the DLK1-MEG3 locus, as being downregulated in islets from humans with T2DM. In this study, using targeted epigenetic modifiers, we prove that increased methylation at the promoter of Meg3 in mouse betaTC6 beta-cells results in decreased transcription of the maternal transcripts associated with this locus. As a result, the sensitivity of beta-cells to cytokine-mediated oxidative stress was increased. Additionally, we demonstrate that an evolutionarily conserved intronic region at the MEG3 locus can function as an enhancer in betaTC6 beta-cells. Using circular chromosome conformation capture followed by high-throughput sequencing, we demonstrate that the promoter of MEG3 physically interacts with this novel enhancer and other putative regulatory elements in this imprinted region in human islets. Remarkably, this enhancer is bound in an allele-specific manner by the transcription factors FOXA2, PDX1, and NKX2.2. Overall, these data suggest that the intronic MEG3 enhancer plays an important role in the regulation of allele-specific expression at the imprinted DLK1-MEG3 locus in human beta-cells, which in turn impacts the sensitivity of beta-cells to cytokine-mediated oxidative stress. CI - (c) 2018 by the American Diabetes Association. FAU - Kameswaran, Vasumathi AU - Kameswaran V AD - Department of Genetics and Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania, Philadelphia, PA. FAU - Golson, Maria L AU - Golson ML AD - Department of Genetics and Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania, Philadelphia, PA. FAU - Ramos-Rodriguez, Mireia AU - Ramos-Rodriguez M AD - Program of Predictive and Personalized Medicine of Cancer, Department of Endocrinology, Germans Trias i Pujol University Hospital and Research Institute, Badalona, Spain. AD - Josep Carreras Leukaemia Research Institute, Badalona, Spain. AD - CIBER de Diabetes y Enfermedades Metabolicas Asociadas, Barcelona, Spain. FAU - Ou, Kristy AU - Ou K AD - Department of Genetics and Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania, Philadelphia, PA. FAU - Wang, Yue J AU - Wang YJ AD - Department of Genetics and Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania, Philadelphia, PA. FAU - Zhang, Jia AU - Zhang J AD - Department of Genetics and Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania, Philadelphia, PA. FAU - Pasquali, Lorenzo AU - Pasquali L AD - Program of Predictive and Personalized Medicine of Cancer, Department of Endocrinology, Germans Trias i Pujol University Hospital and Research Institute, Badalona, Spain. AD - Josep Carreras Leukaemia Research Institute, Badalona, Spain. AD - CIBER de Diabetes y Enfermedades Metabolicas Asociadas, Barcelona, Spain. FAU - Kaestner, Klaus H AU - Kaestner KH AUID- ORCID: 0000-0002-1228-021X AD - Department of Genetics and Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania, Philadelphia, PA kaestner@pennmedicine.upenn.edu. LA - eng GR - P30 DK019525/DK/NIDDK NIH HHS/United States GR - R01 DK088383/DK/NIDDK NIH HHS/United States GR - UC4 DK104119/DK/NIDDK NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20180703 PL - United States TA - Diabetes JT - Diabetes JID - 0372763 RN - 0 (Calcium-Binding Proteins) RN - 0 (Cytokines) RN - 0 (DLK1 protein, human) RN - 0 (Homeobox Protein Nkx-2.2) RN - 0 (Homeodomain Proteins) RN - 0 (Intercellular Signaling Peptides and Proteins) RN - 0 (MEG3 non-coding RNA, human) RN - 0 (Membrane Proteins) RN - 0 (NKX2-2 protein, human) RN - 0 (Nkx2-2 protein, mouse) RN - 0 (Nuclear Proteins) RN - 0 (RNA, Long Noncoding) RN - 0 (Recombinant Fusion Proteins) RN - 0 (Repressor Proteins) RN - 0 (Tgif1 protein, mouse) RN - 0 (Transcription Factors) RN - 147336-22-9 (Green Fluorescent Proteins) RN - EC 2.1.1.37 (DNA (Cytosine-5-)-Methyltransferase 1) SB - IM MH - Animals MH - Calcium-Binding Proteins MH - Cell Line MH - Cytokines/metabolism MH - DNA (Cytosine-5-)-Methyltransferase 1/chemistry/genetics/metabolism MH - *DNA Methylation MH - Diabetes Mellitus, Type 2/*metabolism/pathology MH - Enhancer Elements, Genetic MH - Epigenesis, Genetic MH - *Gene Expression Regulation MH - Genetic Loci MH - Green Fluorescent Proteins/chemistry/genetics/metabolism MH - Homeobox Protein Nkx-2.2 MH - Homeodomain Proteins/chemistry/genetics/metabolism MH - Humans MH - Intercellular Signaling Peptides and Proteins/genetics/*metabolism MH - Islets of Langerhans/*metabolism/pathology MH - Locus Control Region MH - Membrane Proteins/genetics/*metabolism MH - Mice MH - Mutation MH - Nuclear Proteins MH - Oxidative Stress/drug effects MH - *Promoter Regions, Genetic MH - RNA, Long Noncoding/*metabolism MH - Recombinant Fusion Proteins/chemistry/metabolism MH - Repressor Proteins/chemistry/genetics/metabolism MH - Tissue Banks MH - Transcription Factors/genetics/metabolism PMC - PMC6110314 EDAT- 2018/08/08 06:00 MHDA- 2018/09/05 06:00 PMCR- 2019/09/01 CRDT- 2018/08/08 06:00 PHST- 2017/06/14 00:00 [received] PHST- 2018/06/18 00:00 [accepted] PHST- 2018/08/08 06:00 [pubmed] PHST- 2018/09/05 06:00 [medline] PHST- 2018/08/08 06:00 [entrez] PHST- 2019/09/01 00:00 [pmc-release] AID - db17-0682 [pii] AID - 0682 [pii] AID - 10.2337/db17-0682 [doi] PST - ppublish SO - Diabetes. 2018 Sep;67(9):1807-1815. doi: 10.2337/db17-0682. Epub 2018 Jul 3.