The role of Hath6, a newly identified shear-stress-responsive transcription factor, in endothelial cell differentiation and function
The role of Hath6, a newly identified shear-stress-responsive transcription factor, in endothelial cell differentiation and function
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DOI：
10.1242/jcs.136358
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发表时间：
2014-04-01
影响因子：
4
通讯作者：
Pei, Xuetao
中科院分区：
文献类型：
--
作者：
Fang, Fang;Wasserman, Scott M.;Pei, Xuetao
剪切应力诱导的内皮细胞分化的关键调节因子大多不清楚。人类无调同源物6（Hath 6或ATOH 8）是一种内皮选择性和剪切应力响应性转录因子。在这项研究中，我们试图阐明Hath 6在胚胎干细胞内皮特化中的作用。在人胚胎干细胞-内皮细胞（hESC-EC）诱导体系中，Hath 6 mRNA的表达与内皮细胞分化同步上调。随后，使用hESC-EC诱导模型和内皮细胞系进行Hath 6的功能获得和功能丧失研究。模拟切应力处理的Hath 6过表达导致CD 45（-）CD 31（+）KDR（+）群体增加、更高的管状结构形成能力和内皮特异性基因表达增加。相反，Hath 6 mRNA的敲低显著降低内皮分化。Hath 6还促进内皮细胞在内皮基因表达、管状结构形成和细胞迁移方面的成熟。我们进一步证明，编码eNOS的基因是Hath 6通过报告系统测定和蛋白质印迹分析的直接目标，并且eNOS的抑制减少hESC-EC分化。这些结果表明，eNOS在连接Hath 6与内皮表型中起关键作用。在斑马鱼和小鼠胚胎中的进一步原位杂交研究表明，Hath 6的同源物参与血管发生和血管生成。这项研究首次证实了Hath 6对人胚胎内皮细胞分化和功能的积极影响。此外，我们提出了一个潜在的信号通路，通过剪切应力刺激内皮细胞分化。
The key regulators of endothelial differentiation that is induced by shear stress are mostly unclear. Human atonal homolog 6 (Hath6 or ATOH8) is an endothelial-selective and shear-stress-responsive transcription factor. In this study, we sought to elucidate the role of Hath6 in the endothelial specification of embryonic stem cells. In a stepwise human embryonic stem cell to endothelial cell (hESC-EC) induction system, Hath6 mRNA was upregulated synchronously with endothelial determination. Subsequently, gain-of-function and loss-of-function studies of Hath6 were performed using the hESC-EC induction model and endothelial cell lines. The overexpression of Hath6, which mimics shear stress treatment, resulted in an increased CD45(-)CD31(+)KDR(+) population, a higher tubular-structure-formation capacity and increased endothelial-specific gene expression. By contrast, the knockdown of Hath6 mRNA markedly decreased endothelial differentiation. Hath6 also facilitated the maturation of endothelial cells in terms of endothelial gene expression, tubular-structure formation and cell migration. We further demonstrated that the gene encoding eNOS is a direct target of Hath6 through a reporter system assay and western blot analysis, and that the inhibition of eNOS diminishes hESC-EC differentiation. These results suggest that eNOS plays a key role in linking Hath6 to the endothelial phenotype. Further in situ hybridization studies in zebrafish and mouse embryos indicated that homologs of Hath6 are involved in vasculogenesis and angiogenesis. This study provides the first confirmation of the positive impact of Hath6 on human embryonic endothelial differentiation and function. Moreover, we present a potential signaling pathway through which shear stress stimulates endothelial differentiation.