对2型糖尿病和非糖尿病供体的人类胰岛进行全基因组DNA甲基化分析,确定了影响胰岛素分泌的候选基因。

Genome-wide DNA methylation analysis of human pancreatic islets from type 2 diabetic and non-diabetic donors identifies candidate genes that influence insulin secretion.

作者信息

Dayeh Tasnim, Volkov Petr, Salö Sofia, Hall Elin, Nilsson Emma, Olsson Anders H, Kirkpatrick Clare L, Wollheim Claes B, Eliasson Lena, Rönn Tina, Bacos Karl, Ling Charlotte

机构信息

Epigenetics and Diabetes, Department of Clinical Sciences, Lund University Diabetes Centre, Lund University, CRC, Malmö, Sweden.

Department of Cell Physiology and Metabolism, University Medical Center, Geneva, Switzerland.

出版信息

PLoS Genet. 2014 Mar 6;10(3):e1004160. doi: 10.1371/journal.pgen.1004160. eCollection 2014 Mar.

Abstract

Impaired insulin secretion is a hallmark of type 2 diabetes (T2D). Epigenetics may affect disease susceptibility. To describe the human methylome in pancreatic islets and determine the epigenetic basis of T2D, we analyzed DNA methylation of 479,927 CpG sites and the transcriptome in pancreatic islets from T2D and non-diabetic donors. We provide a detailed map of the global DNA methylation pattern in human islets, β- and α-cells. Genomic regions close to the transcription start site showed low degrees of methylation and regions further away from the transcription start site such as the gene body, 3'UTR and intergenic regions showed a higher degree of methylation. While CpG islands were hypomethylated, the surrounding 2 kb shores showed an intermediate degree of methylation, whereas regions further away (shelves and open sea) were hypermethylated in human islets, β- and α-cells. We identified 1,649 CpG sites and 853 genes, including TCF7L2, FTO and KCNQ1, with differential DNA methylation in T2D islets after correction for multiple testing. The majority of the differentially methylated CpG sites had an intermediate degree of methylation and were underrepresented in CpG islands (∼ 7%) and overrepresented in the open sea (∼ 60%). 102 of the differentially methylated genes, including CDKN1A, PDE7B, SEPT9 and EXOC3L2, were differentially expressed in T2D islets. Methylation of CDKN1A and PDE7B promoters in vitro suppressed their transcriptional activity. Functional analyses demonstrated that identified candidate genes affect pancreatic β- and α-cells as Exoc3l silencing reduced exocytosis and overexpression of Cdkn1a, Pde7b and Sept9 perturbed insulin and glucagon secretion in clonal β- and α-cells, respectively. Together, our data can serve as a reference methylome in human islets. We provide new target genes with altered DNA methylation and expression in human T2D islets that contribute to perturbed insulin and glucagon secretion. These results highlight the importance of epigenetics in the pathogenesis of T2D.

摘要

胰岛素分泌受损是2型糖尿病(T2D)的一个标志。表观遗传学可能影响疾病易感性。为了描述人类胰岛中的甲基化组并确定T2D的表观遗传基础,我们分析了来自T2D和非糖尿病供体的胰岛中479,927个CpG位点的DNA甲基化和转录组。我们提供了人类胰岛、β细胞和α细胞中全局DNA甲基化模式的详细图谱。靠近转录起始位点的基因组区域显示出低甲基化程度,而远离转录起始位点的区域,如基因体、3'UTR和基因间区域则显示出较高的甲基化程度。虽然CpG岛是低甲基化的,但周围2 kb的海岸显示出中等程度的甲基化,而在人类胰岛、β细胞和α细胞中,更远的区域(陆架和公海)是高甲基化的。在进行多重检验校正后,我们在T2D胰岛中鉴定出1,649个CpG位点和853个基因,包括TCF7L2、FTO和KCNQ1,它们具有差异DNA甲基化。大多数差异甲基化的CpG位点具有中等程度的甲基化,在CpG岛中占比不足(约7%),在公海中占比过高(约60%)。102个差异甲基化基因,包括CDKN1A、PDE7B、SEPT9和EXOC3L2,在T2D胰岛中差异表达。体外CDKN1A和PDE7B启动子的甲基化抑制了它们的转录活性。功能分析表明,鉴定出的候选基因会影响胰腺β细胞和α细胞,因为Exoc3l沉默会减少胞吐作用,而Cdkn1a、Pde7b和Sept9的过表达分别会干扰克隆β细胞和α细胞中的胰岛素和胰高血糖素分泌。总之,我们的数据可作为人类胰岛中的参考甲基化组。我们提供了在人类T2D胰岛中DNA甲基化和表达发生改变的新靶基因,这些基因导致胰岛素和胰高血糖素分泌紊乱。这些结果突出了表观遗传学在T2D发病机制中的重要性。

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