Meganathan K, Jagtap S, Srinivasan S P, Wagh V, Hescheler J, Hengstler J, Leist M, Sachinidis A
Center of Physiology and Pathophysiology, Institute of Neurophysiology and Center for Molecular Medicine Cologne (CMMC), Robert-Koch-Str. 39, Cologne, Germany.
Leibniz Research Centre for Working Environment and Human Factors, Technical University of Dortmund (IfADo), Dortmund, Germany.
Cell Death Dis. 2015 May 7;6(5):e1756. doi: 10.1038/cddis.2015.121.
Human embryonic stem cells (hESCs) may be applied to develop human-relevant sensitive in vitro test systems for monitoring developmental toxicants. The aim of this study was to identify potential developmental toxicity mechanisms of the histone deacetylase inhibitors (HDAC) valproic acid (VPA), suberoylanilide hydroxamic acid (SAHA) and trichostatin A (TSA) relevant to the in vivo condition using a hESC model in combination with specific differentiation protocols and genome-wide gene expression and microRNA profiling. Analysis of the gene expression data showed that VPA repressed neural tube and dorsal forebrain (OTX2, ISL1, EMX2 and SOX10)-related transcripts. In addition, VPA upregulates axonogenesis and ventral forebrain-associated genes, such as SLIT1, SEMA3A, DLX2/4 and GAD2. HDACi-induced expression of miR-378 and knockdown of miR-378 increases the expression of OTX2 and EMX2, which supports our hypothesis that HDACi targets forebrain markers through miR-378. In conclusion, multilineage differentiation in vitro test system is very sensitive for monitoring molecular activities relevant to in vivo neuronal developmental toxicity. Moreover, miR-378 seems to repress the expression of the OTX2 and EMX2 and therefore could be a regulator of the development of neural tube and dorsal forebrain neurons.
人类胚胎干细胞(hESCs)可用于开发与人类相关的敏感体外测试系统,以监测发育毒性物质。本研究的目的是使用hESC模型,结合特定的分化方案以及全基因组基因表达和微小RNA分析,确定组蛋白去乙酰化酶抑制剂(HDAC)丙戊酸(VPA)、辛二酰苯胺异羟肟酸(SAHA)和曲古抑菌素A(TSA)在体内条件下潜在的发育毒性机制。基因表达数据分析表明,VPA抑制神经管和背侧前脑(OTX2、ISL1、EMX2和SOX10)相关转录本。此外,VPA上调轴突发生和腹侧前脑相关基因的表达,如SLIT1、SEMA3A、DLX2/4和GAD2。HDACi诱导的miR-378表达以及miR-378的敲低增加了OTX2和EMX2的表达,这支持了我们的假设,即HDACi通过miR-378靶向作用于前脑标志物。总之,体外多谱系分化测试系统对于监测与体内神经元发育毒性相关的分子活性非常敏感。此外,miR-378似乎抑制OTX2和EMX2的表达,因此可能是神经管和背侧前脑神经元发育的调节因子。