Abnormal epigenetic changes during differentiation of human skeletal muscle stem cells from obese subjects.
Abnormal epigenetic changes during differentiation of human skeletal muscle stem cells from obese subjects.
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DOI：
10.1186/s12916-017-0792-x
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发表时间：
2017-02-22
期刊：
影响因子：
9.3
通讯作者：
Ling C
中科院分区：
文献类型：
--
作者：
Davegårdh C;Broholm C;Perfilyev A;Henriksen T;García-Calzón S;Peijs L;Hansen NS;Volkov P;Kjøbsted R;Wojtaszewski JF;Pedersen M;Pedersen BK;Ballak DB;Dinarello CA;Heinhuis B;Joosten LA;Nilsson E;Vaag A;Scheele C;Ling C
人骨骼肌干细胞对肌肉再生非常重要。然而，在成人肌发生过程中发生的全基因组DNA甲基化和表达变化尚未详细描述，可能会发现新的肌发生因子。此外，肥胖与低相对肌肉质量和降低的代谢有关。发生在肌肉发生过程中的表观遗传改变可能有助于这些缺陷。我们使用Infinium HumanMethylation 450 BeadChip Kit（Illumina）和HumanHT-12 Expression BeadChip（Illumina）分析了来自14名非肥胖和14名肥胖个体的原代人成肌细胞分化前后的全基因组DNA甲基化和转录。在原代人成肌细胞和转基因小鼠模型中使用siRNA介导的基因沉默进行功能随访实验。我们观察了原代人肌肉干细胞（成肌细胞）分化过程中DNA甲基化和表达模式的全基因组变化。我们鉴定了肌生成转录因子（MYOD 1、MYOG、MYF 5、MYF 6、PAX 7、MEF 2A、MEF 2C和MEF 2D）、细胞周期调节因子、代谢酶和以前与肌生成无关的基因（包括IL 32、金属硫蛋白和妊娠特异性β-1-糖蛋白）的表观遗传和转录变化。功能研究表明，IL-32作为一种新的靶点，可调节人体肌肉发生、胰岛素敏感性和肌细胞中的ATP水平。此外，IL 32转基因小鼠具有降低的胰岛素反应和肌肉重量。值得注意的是，在肥胖与非肥胖受试者的成肌细胞分化过程中观察到大约3.7倍的甲基化变化（147，161对39，572）。与此相一致，DNMT 1的表达在肌生成过程中仅在肥胖受试者中增加。有趣的是，许多与代谢疾病和表观遗传调控有关的基因仅在肥胖受试者的分化过程中表现出差异甲基化和表达。我们的研究将IL-32鉴定为一种新的肌源性调节因子，提供了人类肌肉干细胞分化过程中动态表观基因组的全面图谱，并揭示了肥胖症的异常表观遗传变化。本文的在线版本（doi：10.1186/s12916-017-0792-x）包含补充材料，可供授权用户使用。
Human skeletal muscle stem cells are important for muscle regeneration. However, the combined genome-wide DNA methylation and expression changes taking place during adult myogenesis have not been described in detail and novel myogenic factors may be discovered. Additionally, obesity is associated with low relative muscle mass and diminished metabolism. Epigenetic alterations taking place during myogenesis might contribute to these defects. We used Infinium HumanMethylation450 BeadChip Kit (Illumina) and HumanHT-12 Expression BeadChip (Illumina) to analyze genome-wide DNA methylation and transcription before versus after differentiation of primary human myoblasts from 14 non-obese and 14 obese individuals. Functional follow-up experiments were performed using siRNA mediated gene silencing in primary human myoblasts and a transgenic mouse model. We observed genome-wide changes in DNA methylation and expression patterns during differentiation of primary human muscle stem cells (myoblasts). We identified epigenetic and transcriptional changes of myogenic transcription factors (MYOD1, MYOG, MYF5, MYF6, PAX7, MEF2A, MEF2C, and MEF2D), cell cycle regulators, metabolic enzymes and genes previously not linked to myogenesis, including IL32, metallothioneins, and pregnancy-specific beta-1-glycoproteins. Functional studies demonstrated IL-32 as a novel target that regulates human myogenesis, insulin sensitivity and ATP levels in muscle cells. Furthermore, IL32 transgenic mice had reduced insulin response and muscle weight. Remarkably, approximately 3.7 times more methylation changes (147,161 versus 39,572) were observed during differentiation of myoblasts from obese versus non-obese subjects. In accordance, DNMT1 expression increased during myogenesis only in obese subjects. Interestingly, numerous genes implicated in metabolic diseases and epigenetic regulation showed differential methylation and expression during differentiation only in obese subjects. Our study identifies IL-32 as a novel myogenic regulator, provides a comprehensive map of the dynamic epigenome during differentiation of human muscle stem cells and reveals abnormal epigenetic changes in obesity. The online version of this article (doi:10.1186/s12916-017-0792-x) contains supplementary material, which is available to authorized users.