Dual small-molecule targeting of SMAD signaling stimulates human induced pluripotent stem cells toward neural lineages.
Dual small-molecule targeting of SMAD signaling stimulates human induced pluripotent stem cells toward neural lineages.
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DOI：
10.1371/journal.pone.0106952
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Issaragrisil S
中科院分区：
文献类型：
--
作者：
Wattanapanitch M;Klincumhom N;Potirat P;Amornpisutt R;Lorthongpanich C;U-pratya Y;Laowtammathron C;Kheolamai P;Poungvarin N;Issaragrisil S
帕金森病 (PD)、亨廷顿舞蹈病 (HD) 和阿尔茨海默病 (AD) 等无法治愈的神经系统疾病非常常见，并且可能危及生命，因为它们的疾病症状不断进展，而治疗选择有限。为了为基于细胞的移植提供替代的可再生细胞来源并作为神经系统疾病的研究模型，我们从人真皮成纤维细胞（HDF）中产生了诱导多能干细胞（iPSC），然后通过双重SMAD信号抑制剂将其分化为神经祖细胞（NPC）和成熟神经元。逆转录病毒转导后，通过补充组蛋白脱乙酰酶抑制剂丙戊酸 (VPA) 和 p160-Rho 相关卷曲螺旋激酶 (ROCK) 抑制剂 Y-27632 提高重编程效率。我们获得了许多 iPS 集落，它们在形态、细胞表面抗原、多能性相关基因和蛋白表达以及体外和体内分化潜力方面与人胚胎干细胞具有相似的特征。经过 Noggin 和 SMAD 信号通路抑制剂 SB431542 处理后，HDF-iPSC 能够快速有效地分化为神经谱系。神经诱导六天后，在贴壁单层培养物中观察到神经上皮细胞（NEPC），其具有进一步分化为 NPC 和神经元的能力，其特征在于其形态以及神经元特异性转录物和蛋白质的表达。我们建议我们的研究可用于生成神经系统疾病患者特异性 iPSC，从而更好地了解疾病发病机制和药物敏感性测定。
Incurable neurological disorders such as Parkinson’s disease (PD), Huntington’s disease (HD), and Alzheimer’s disease (AD) are very common and can be life-threatening because of their progressive disease symptoms with limited treatment options. To provide an alternative renewable cell source for cell-based transplantation and as study models for neurological diseases, we generated induced pluripotent stem cells (iPSCs) from human dermal fibroblasts (HDFs) and then differentiated them into neural progenitor cells (NPCs) and mature neurons by dual SMAD signaling inhibitors. Reprogramming efficiency was improved by supplementing the histone deacethylase inhibitor, valproic acid (VPA), and inhibitor of p160-Rho associated coiled-coil kinase (ROCK), Y-27632, after retroviral transduction. We obtained a number of iPS colonies that shared similar characteristics with human embryonic stem cells in terms of their morphology, cell surface antigens, pluripotency-associated gene and protein expressions as well as their in vitro and in vivo differentiation potentials. After treatment with Noggin and SB431542, inhibitors of the SMAD signaling pathway, HDF-iPSCs demonstrated rapid and efficient differentiation into neural lineages. Six days after neural induction, neuroepithelial cells (NEPCs) were observed in the adherent monolayer culture, which had the ability to differentiate further into NPCs and neurons, as characterized by their morphology and the expression of neuron-specific transcripts and proteins. We propose that our study may be applied to generate neurological disease patient-specific iPSCs allowing better understanding of disease pathogenesis and drug sensitivity assays.
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