Efficient Generation of Dopamine Neuron-Like Cells From Skin-Derived Precursors With a Synthetic Peptide Derived From von Hippel-Lindau Protein
Efficient Generation of Dopamine Neuron-Like Cells From Skin-Derived Precursors With a Synthetic Peptide Derived From von Hippel-Lindau Protein
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DOI：
10.1089/scd.2008.0379
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发表时间：
2009-12-01
影响因子：
4
通讯作者：
Kanno, Hiroshi
中科院分区：
文献类型：
--
作者：
Kubo, Atsuhiko;Yoshida, Tetsuhiko;Kanno, Hiroshi
来自哺乳动物真皮的皮肤源性前体细胞（SKPs）代表能够分化为神经和中胚层前体的神经嵴相关干细胞。SKPs具有临床意义，因为它们可作为可获得的自体供体细胞用于神经元难治性疾病的神经元修复。然而，很少有人知道从SKPs的神经元的有效生成，从SKPs产生的神经元的表型已受到限制。此外，利用其产生的神经元作为供体细胞的神经元修复尚未实现。von Hippel-Lindau蛋白（pVHL）是在神经元分化过程中起重要作用的蛋白质之一，并且最近已经证明了通过细胞内递送衍生自延伸蛋白BC结合位点的合成VHL肽来使神经祖细胞分化为神经元。在本研究中，一个合成的VHL肽衍生自延伸蛋白BC结合位点缀合到HIV-TAT蛋白（TATVHL肽）的蛋白转导结构域（PTD），以促进进入细胞，我们证明了有效的生产与多巴胺能表型的细胞SKPs与TATVHL肽的细胞内交付，并确定了所产生的细胞。TATVHL肽处理的SKPs表达神经元标志物蛋白，特别是多巴胺神经元标志物，并且还上调前神经碱性螺旋-环-螺旋因子的mRNA水平。在TATVHL肽治疗后，将SKPs移植到帕金森病（PD）模型大鼠中，在PD模型大鼠中充分分化为多巴胺神经元样细胞，并且部分但显著地校正了PD模型大鼠的行为。所产生的多巴胺神经元样细胞有望作为PD神经元修复的供体细胞。
Skin-derived precursors (SKPs) from mammalian dermis represent neural crest-related stem cells capable of differentiating into both neural and mesodermal progency. SKPs are of clinical interest because they serve as accessible autologous donor cells for neuronal repair for neuronal intractable diseases. However, little is known about the efficient generation of neurons from SKPs, and phenotypes of neurons generated from SKPs have been restricted. In addition, the neuronal repair using their generated neurons as donor cells has not been achieved. The von Hippel-Lindau protein (pVHL) is one of the proteins that play an important role during neuronal differentiation, and recently neuronal differentiation of neural progenitor cells by intracellular delivery of a synthetic VHL peptide derived from elongin BC-binding site has been demonstrated. In the present study, a synthetic VHL peptide derived from elongin BC-binding site was conjugated to the protein transduction domain (PTD) of HIV-TAT protein (TATVHL peptide) to facilitate entry into cells, and we demonstrate the efficient generation of cells with dopaminergic phenotype from SKPs with the intracellular delivery of TATVHL peptide, and characterized the generated cells. The TATVHL peptide-treated SKPs expressed neuronal marker proteins, particularly dopamine neuron markers, and also up-regulated mRNA levels of proneural basic helix-loop-helix factors. After the TATVHL peptide treatment, transplanted SKPs into Parkinson's disease (PD) model rats sufficiently differentiated into dopamine neuron-like cells in PD model rats, and partially but significantly corrected behavior of PD model rats. The generated dopamine neuron-like cells are expected to serve as donor cells for neuronal repair for PD.