Cell cycle and p53 gate the direct conversion of human fibroblasts to dopaminergic neurons.
Cell cycle and p53 gate the direct conversion of human fibroblasts to dopaminergic neurons.
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DOI：
10.1038/ncomms10100
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发表时间：
2015-12-07
影响因子：
16.6
通讯作者：
Feng J
中科院分区：
文献类型：
--
作者：
Jiang H;Xu Z;Zhong P;Ren Y;Liang G;Schilling HA;Hu Z;Zhang Y;Wang X;Chen S;Yan Z;Feng J
成纤维细胞向诱导的多巴胺能（iDA）神经元和其他细胞类型的直接转化证明了细胞命运的可塑性。这些相对快速的转化的低效率表明存在动力学障碍以保护细胞类型身份。在这里，我们表明，抑制p53，结合细胞周期停滞在G1期和适当的细胞外环境，显着提高效率的人成纤维细胞转分化为iDA神经元的Ascl1，Nurr1，Lmx1a和miR124。这种转化依赖于Tet1，因为G1阻滞、p53敲低或重编程因子的表达协同诱导Tet1。Tet1敲除废除转分化，而其过表达增强转换。iDA神经元表达中脑DA神经元的标记物，并具有活跃的多巴胺能传递。我们的研究结果表明，克服这些动力学障碍可能使高效的表观遗传重编程一般，并将产生患者特异性中脑DA神经元帕金森病的研究和治疗。 成纤维细胞重编程为诱导的多巴胺能神经元的效率通常较低。在此，Jiang等证明了通过诱导细胞周期停滞、抑制p53、泰特1和通过向培养基中添加神经营养因子来提高转分化率。
The direct conversion of fibroblasts to induced dopaminergic (iDA) neurons and other cell types demonstrates the plasticity of cell fate. The low efficiency of these relatively fast conversions suggests that kinetic barriers exist to safeguard cell-type identity. Here we show that suppression of p53, in conjunction with cell cycle arrest at G1 and appropriate extracellular environment, markedly increase the efficiency in the transdifferentiation of human fibroblasts to iDA neurons by Ascl1, Nurr1, Lmx1a and miR124. The conversion is dependent on Tet1, as G1 arrest, p53 knockdown or expression of the reprogramming factors induces Tet1 synergistically. Tet1 knockdown abolishes the transdifferentiation while its overexpression enhances the conversion. The iDA neurons express markers for midbrain DA neurons and have active dopaminergic transmission. Our results suggest that overcoming these kinetic barriers may enable highly efficient epigenetic reprogramming in general and will generate patient-specific midbrain DA neurons for Parkinson's disease research and therapy. The efficiency of reprogramming of fibroblasts into induced dopaminergic neurons is often low. Here, Jiang et al. demonstrate increased transdifferentiation rates by inducing cell cycle arrest, suppressing p53, Tet 1 and by adding neurotrophic factors to the media.