Sagal Jonathan, Zhan Xiping, Xu Jinchong, Tilghman Jessica, Karuppagounder Senthilkumar S, Chen Li, Dawson Valina L, Dawson Ted M, Laterra John, Ying Mingyao
Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA.
Hugo W. Moser Research Institute at Kennedy Krieger, Baltimore, Maryland, USA; Department of Physiology and Biophysics, Howard University, Washington, D.C., USA; Department of Neurology, Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Department of Neuroscience, Department of Physiology, and Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Adrienne Helis Malvin Medical Research Foundation, New Orleans, Louisiana, USA
Stem Cells Transl Med. 2014 Aug;3(8):888-98. doi: 10.5966/sctm.2013-0213. Epub 2014 Jun 5.
Human pluripotent stem cells (PSCs) are a promising cell resource for various applications in regenerative medicine. Highly efficient approaches that differentiate human PSCs into functional lineage-specific neurons are critical for modeling neurological disorders and testing potential therapies. Proneural transcription factors are crucial drivers of neuron development and hold promise for driving highly efficient neuronal conversion in PSCs. Here, we study the functions of proneural transcription factor Atoh1 in the neuronal differentiation of PSCs. We show that Atoh1 is induced during the neuronal conversion of PSCs and that ectopic Atoh1 expression is sufficient to drive PSCs into neurons with high efficiency. Atoh1 induction, in combination with cell extrinsic factors, differentiates PSCs into functional dopaminergic (DA) neurons with >80% purity. Atoh1-induced DA neurons recapitulate key biochemical and electrophysiological features of midbrain DA neurons, the degeneration of which is responsible for clinical symptoms in Parkinson's disease (PD). Atoh1-induced DA neurons provide a reliable disease model for studying PD pathogenesis, such as neurotoxin-induced neurodegeneration in PD. Overall, our results determine the role of Atoh1 in regulating neuronal differentiation and neuron subtype specification of human PSCs. Our Atoh1-mediated differentiation approach will enable large-scale applications of PD patient-derived midbrain DA neurons in mechanistic studies and drug screening for both familial and sporadic PD.
人类多能干细胞（PSCs）是再生医学中各种应用的一种有前景的细胞资源。将人类PSCs高效分化为功能性谱系特异性神经元的方法对于模拟神经疾病和测试潜在治疗方法至关重要。原神经转录因子是神经元发育的关键驱动因素，有望推动PSCs中高效的神经元转化。在此，我们研究原神经转录因子Atoh1在PSCs神经元分化中的功能。我们表明，Atoh1在PSCs的神经元转化过程中被诱导，并且异位表达Atoh1足以高效地将PSCs驱动为神经元。Atoh1诱导与细胞外源性因子相结合，可将PSCs分化为纯度>80%的功能性多巴胺能（DA）神经元。Atoh1诱导的DA神经元重现了中脑DA神经元的关键生化和电生理特征，中脑DA神经元的退化是帕金森病（PD）临床症状的原因。Atoh1诱导的DA神经元为研究PD发病机制提供了一个可靠的疾病模型，例如PD中神经毒素诱导的神经变性。总体而言，我们的结果确定了Atoh1在调节人类PSCs神经元分化和神经元亚型特异性中的作用。我们的Atoh1介导的分化方法将使源自PD患者的中脑DA神经元能够大规模应用于家族性和散发性PD的机制研究和药物筛选。
