Epigenetic histone modification and cardiovascular lineage programming in mouse embryonic stem cells exposed to laminar shear stress
Epigenetic histone modification and cardiovascular lineage programming in mouse embryonic stem cells exposed to laminar shear stress
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DOI：
10.1161/01.res.0000159181.06379.63
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发表时间：
2005-03-18
影响因子：
20.1
通讯作者：
Gaetano, C
中科院分区：
文献类型：
--
作者：
Illi, B;Scopece, A;Gaetano, C
实验表明，剪切应力在小鼠和斑马鱼胚胎心脏发育过程中具有形态发生作用。然而，这种效应的分子基础仍然难以捉摸。我们之前的工作描述了在成人内皮细胞中，SS通过诱导组蛋白的表观遗传修饰和激活具有乙酰转移酶活性的转录复合物来调节基因表达。在这项研究中，我们评估了SS处理是否可以表观遗传修饰组蛋白并影响小鼠胚胎干细胞（ES）的细胞分化。细胞暴露于10达因/ cm(2)/s（-1）的层流SS中，或在存在或不存在组蛋白去乙酰化酶抑制剂trichostatin a （TSA）的静态条件下保存。这些实验表明，SS增强了14位组蛋白H3的赖氨酸乙酰化（K14）、10位丝氨酸磷酸化（S10）和79位赖氨酸甲基化（K79），并与TSA协同作用，诱导组蛋白H4乙酰化、组蛋白H3的S10和K14磷酸化乙酰化。此外，暴露于SS的ES细胞强烈激活了血管内皮生长因子（VEGF）受体2启动子的转录。这种效果与心血管标志物的早期诱导相似，包括平滑肌肌动蛋白、平滑肌蛋白22- α、血小板内皮细胞粘附分子-1、VEGF受体2、肌细胞增强因子- 2c （MEF2C）和α -肌动蛋白。在这种条件下，可以从ss处理的ES细胞中分离到转录因子MEF2C和Sma/MAD同源蛋白4，这些转录因子与cAMP反应元件结合蛋白乙酰转移酶结合。这些结果为小鼠胚胎干细胞依赖ss的心血管承诺提供了分子基础，并表明层流可能成功地应用于体外心血管前体的生产。
Experimental evidence indicates that shear stress (SS) exerts a morphogenetic function during cardiac development of mouse and zebrafish embryos. However, the molecular basis for this effect is still elusive. Our previous work described that in adult endothelial cells, SS regulates gene expression by inducing epigenetic modification of histones and activation of transcription complexes bearing acetyltransferase activity. In this study, we evaluated whether SS treatment could epigenetically modify histones and influence cell differentiation in mouse embryonic stem (ES) cells. Cells were exposed to a laminar SS of 10 dyne per cm(2)/s(-1), or kept in static conditions in the presence or absence of the histone deacetylase inhibitor trichostatin A (TSA). These experiments revealed that SS enhanced lysine acetylation of histone H3 at position 14 (K14), as well as serine phosphorylation at position 10 (S10) and lysine methylation at position 79 (K79), and cooperated with TSA, inducing acetylation of histone H4 and phosphoacetylation of S10 and K14 of histone H3. In addition, ES cells exposed to SS strongly activated transcription from the vascular endothelial growth factor (VEGF) receptor 2 promoter. This effect was paralleled by an early induction of cardiovascular markers, including smooth muscle actin, smooth muscle protein 22-alpha, platelet-endothelial cell adhesion molecule-1, VEGF receptor 2, myocyte enhancer factor-2C (MEF2C), and alpha-sarcomeric actin. In this condition, transcription factors MEF2C and Sma/MAD homolog protein 4 could be isolated from SS-treated ES cells complexed with the cAMP response element-binding protein acetyltransferase. These results provide molecular basis for the SS-dependent cardiovascular commitment of mouse ES cells and suggest that laminar flow may be successfully applied for the in vitro production of cardiovascular precursors.