The redox function of APE1 is involved in the differentiation process of stem cells toward a neuronal cell fate.
The redox function of APE1 is involved in the differentiation process of stem cells toward a neuronal cell fate.
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DOI：
10.1371/journal.pone.0089232
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Beltrami AP
中科院分区：
文献类型：
--
作者：
Domenis R;Bergamin N;Gianfranceschi G;Vascotto C;Romanello M;Rigo S;Vagnarelli G;Faggiani M;Parodi P;Kelley MR;Beltrami CA;Cesselli D;Tell G;Beltrami AP
低至中等水平的活性氧（ROS）通过分子途径控制神经发生的不同步骤，这些分子途径仅部分被解密。虽然已经假设氧化还原敏感性分子参与神经元分化，但这一过程的分子基础尚未阐明。因此，本工作的目的是研究氧化还原敏感的多功能蛋白APE 1/Ref-1（APE 1）在人脂肪组织来源的多能成体干细胞（hAT-MASC）和胚胎癌干细胞（EC）向神经元表型分化过程中所起的作用。方法和结果：应用一个确定的方案，hAT-MASC可以采取神经命运。在此成熟过程中，分化细胞显著增加其细胞内活性氧（ROS）水平，并增加与染色质结合的APE 1核分数。后一种情况被核NF-κB的增加所阻止，核NF-κ B是一种由APE 1以氧化还原依赖性方式调节的转录因子。重要的是，向分化培养基中添加抗氧化剂N-乙酰半胱氨酸（NAC）部分地防止了APE 1的核积累，增加了hAT-MASC的神经元分化。为了研究APE 1在分化过程中的参与，我们采用了APE 1氧化还原功能的特异性抑制剂E3330。向神经源性胚胎癌细胞系NT 2-D1或hAT-MASC中添加E3330增加了干细胞向神经表型的分化，使分化偏向特定亚型，如多巴胺能细胞。总之，在干细胞向神经外胚层表型分化的过程中，APE 1以ROS依赖的方式被募集到染色质中。该事件与APE 1对神经发生的抑制作用相关，该抑制作用可被E3330逆转。因此，E3330可用于促进神经分化和使干细胞的分化潜能偏向特定神经元亚型。这些发现为氧化还原介导的神经元分化程序假说提供了分子基础。
Low-to-moderate levels of reactive oxygen species (ROS) govern different steps of neurogenesis via molecular pathways that have been decrypted only partially. Although it has been postulated that redox-sensitive molecules are involved in neuronal differentiation, the molecular bases for this process have not been elucidated yet. The aim of this work was therefore to study the role played by the redox-sensitive, multifunctional protein APE1/Ref-1 (APE1) in the differentiation process of human adipose tissue-derived multipotent adult stem cells (hAT-MASC) and embryonic carcinoma stem cells (EC) towards a neuronal phenotype. Methods and results: Applying a definite protocol, hAT-MASC can adopt a neural fate. During this maturation process, differentiating cells significantly increase their intracellular Reactive Oxygen Species (ROS) levels and increase the APE1 nuclear fraction bound to chromatin. This latter event is paralleled by the increase of nuclear NF-κB, a transcription factor regulated by APE1 in a redox-dependent fashion. Importantly, the addition of the antioxidant N-acetyl cysteine (NAC) to the differentiation medium partially prevents the nuclear accumulation of APE1, increasing the neuronal differentiation of hAT-MASC. To investigate the involvement of APE1 in the differentiation process, we employed E3330, a specific inhibitor of the APE1 redox function. The addition of E3330, either to the neurogenic embryonic carcinoma cell line NT2-D1or to hAT-MASC, increases the differentiation of stem cells towards a neural phenotype, biasing the differentiation towards specific subtypes, such as dopaminergic cells. In conclusion, during the differentiation process of stem cells towards a neuroectodermic phenotype, APE1 is recruited, in a ROS-dependent manner, to the chromatin. This event is associated with an inhibitory effect of APE1 on neurogenesis that may be reversed by E3330. Therefore, E3330 may be employed both to boost neural differentiation and to bias the differentiation potential of stem cells towards specific neuronal subtypes. These findings provide a molecular basis for the redox-mediated hypothesis of neuronal differentiation program.
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