Exploring the effect of epigenetic modifiers on developing insulin-secreting cells
Exploring the effect of epigenetic modifiers on developing insulin-secreting cells
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DOI：
10.1007/s13577-019-00292-y
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发表时间：
2019-11-21
期刊：
影响因子：
4.3
通讯作者：
El-Serafi, Ahmed T.
中科院分区：
文献类型：
--
作者：
Elsharkawi, Ibrahim;Parambath, Divyasree;El-Serafi, Ahmed T.
糖尿病是一个世界性的健康问题，发病率不断上升。目前的管理方式未能成功减少合并症。这项研究旨在通过表观遗传修饰方法改善间充质基质细胞（MSC）向葡萄糖敏感、胰岛素分泌细胞的分化，从而增强糖尿病的再生解决方案。使用表观遗传修饰剂的 3 天治疗方案，地西他滨（5-aza-2'-脱氧胞苷；Aza）； DNA甲基化抑制剂或伏立诺他（辛二酰苯胺异羟肟酸；SAHA）；将组蛋白脱乙酰酶抑制剂添加到两种不同的人类干细胞系中。这些细胞遵循多步分化方案，该方案在时间方法中提供了关键的触发因素。 Aza 预处理组显示出较高的细胞内胰岛素和转录因子“PDX-1”表达。 ELISA 表明，细胞通过在培养基中分泌胰岛素来应对高葡萄糖挑战。基因表达显示胰岛素、葡萄糖转运蛋白 2、葡萄糖激酶以及转录因子 MafA 和 NKX6.1 基因的诱导。尽管 SAHA 显示胰岛素分泌上调，但与对照相比，细胞无法对高葡萄糖挑战做出反应。有趣的是，Aza 处理的细胞在培养结束时表现出整体 DNA 甲基化水平显着下降。总之，Aza 的这一额外步骤可以增强 MSC 对胰岛素分泌细胞经典分化方案的反应，并可能有助于为糖尿病患者建立再生解决方案。
Diabetes is a worldwide health problem with increasing incidence. The current management modalities did not succeed to decrease comorbidities. This study aimed at enhancing the regenerative solution for diabetes by improving the differentiation of mesenchymal stromal cells (MSC) into glucose-sensitive, insulin-secreting cells through an epigenetic modification approach. A 3-day treatment protocol with the epigenetic modifiers, either decitabine (5-aza-2 '-deoxycytidine; Aza); a DNA methylation inhibitor or Vorinostat (suberoylanilide hydroxamic acid; SAHA); a histone deacetylase inhibitor was added to two different human stem cell lines. The cells followed a multi-step differentiation protocol that provided the critical triggers in a temporal approach. Aza-pretreated group showed higher intracellular expression of insulin and the transcription factor 'PDX-1'. The cells responded to the high glucose challenge by secreting insulin in the media, as shown by ELISA. Gene expression showed induction of the genes for insulin, the glucose transporter 2, glucokinase, as well as the transcription factors MafA and NKX6.1. Although SAHA showed upregulation of insulin secretion, in comparison to control, the cells could not respond to the high glucose challenge. Interestingly, Aza-treated cells showed a significant decrease in the global DNA methylation level at the end of the culture. In conclusion, this additional step with Aza could enhance the response of MSC to the classical differentiation protocol for insulin-secreting cells and may help in establishing a regenerative solution for patients with diabetes.