Department of Cell Biology, MOE Key Laboratory of Geriatric Diseases and Immunology, Suzhou Medical College of Soochow University, Suzhou 215123, China.
Department of Orthopedics, Suzhou Yongding Hospital, Suzhou 215200, China.
Neurobiol Dis. 2024 Oct 15;201:106683. doi: 10.1016/j.nbd.2024.106683. Epub 2024 Sep 27.
Parkinson's disease (PD) is a neurodegenerative disorder characterized by motor deficits due to the depletion of nigrostriatal dopamine. Stem cell differentiation therapy emerges as a promising treatment option for sustained symptom relief. In this study, we successfully developed a one-step differentiation system using the YFBP cocktail (Y27632, Forskolin, SB431542, and SP600125) to effectively convert human umbilical cord mesenchymal stem cells (hUCMSCs) into dopaminergic neurons without genetic modification. This approach addresses the challenge of rapidly and safely generating functional neurons on a large scale. After a 7-day induction period, over 80 % of the cells were double-positive for TUBB3 and NEUN. Transcriptome analysis revealed the dual roles of the cocktail in inducing fate erasure in mesenchymal stem cells and activating the neuronal program. Notably, these chemically induced cells (CiNs) did not express HLA class II genes, preserving their immune-privileged status. Further study indicated that YFBP significantly downregulated p53 signaling and accelerated the differentiation process when Pifithrin-α, a p53 signaling inhibitor, was applied. Additionally, Wnt/β-catenin signaling was transiently activated within one day, but the prolonged activation hindered the neuronal differentiation of hUCMSCs. Upon transplantation into the striatum of mice, CiNs survived well and tested positive for dopaminergic neuron markers. They exhibited typical action potentials and sodium and potassium ion channel activity, demonstrating neuronal electrophysiological activity. Furthermore, CiNs treatment significantly increased the number of tyrosine hydroxylase-positive cells and the concentration of dopamine in the striatum, effectively ameliorating movement disorders in PD mice. Overall, our study provides a secure and reliable framework for cell replacement therapy for Parkinson's disease.
帕金森病（PD）是一种神经退行性疾病，其特征是由于黑质纹状体多巴胺耗竭而导致运动功能障碍。干细胞分化治疗作为一种有前途的治疗选择，可实现持续的症状缓解。在这项研究中，我们成功开发了一种使用 YFBP 鸡尾酒（Y27632、Forskolin、SB431542 和 SP600125）的一步分化系统，无需基因修饰即可有效地将人脐带间充质干细胞（hUCMSCs）分化为多巴胺能神经元。这种方法解决了快速、安全地大规模生成功能性神经元的挑战。经过 7 天的诱导期，超过 80%的细胞对 TUBB3 和 NEUN 双阳性。转录组分析揭示了鸡尾酒在诱导间充质干细胞命运消除和激活神经元程序中的双重作用。值得注意的是，这些化学诱导的细胞（CiNs）不表达 HLA Ⅱ类基因，保持其免疫特权状态。进一步的研究表明，YFBP 显著下调了 p53 信号通路，并且当应用 p53 信号通路抑制剂 Pifithrin-α时，加速了分化过程。此外，Wnt/β-catenin 信号通路在一天内被短暂激活，但长期激活会阻碍 hUCMSCs 的神经元分化。CiNs 移植到小鼠纹状体后，存活良好，并对多巴胺能神经元标志物呈阳性。它们表现出典型的动作电位和钠钾离子通道活性，证明具有神经元电生理活性。此外，CiNs 治疗显著增加了酪氨酸羟化酶阳性细胞的数量和纹状体中多巴胺的浓度，有效改善了 PD 小鼠的运动障碍。总体而言，我们的研究为帕金森病的细胞替代治疗提供了安全可靠的框架。
