Modeling the phenotype of spinal muscular atrophy by the direct conversion of human fibroblasts to motor neurons.
Modeling the phenotype of spinal muscular atrophy by the direct conversion of human fibroblasts to motor neurons.
复制标题
通过将人成纤维细胞直接转化为运动神经元来模拟脊髓性肌萎缩症的表型
DOI：
10.18632/oncotarget.14641
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发表时间：
2017-02-14
期刊：
影响因子：
--
通讯作者：
Chen WJ
中科院分区：
文献类型：
--
作者：
Zhang QJ;Li JJ;Lin X;Lu YQ;Guo XX;Dong EL;Zhao M;He J;Wang N;Chen WJ
脊髓性肌萎缩症(SMA)是一种以脊髓运动神经元选择性变性为特征的致死性常染色体隐性遗传性神经系统疾病。近年来，细胞重编程技术的发展为体外获得患者特异性神经元提供了另一种有效的方法。在本研究中，我们将这项技术应用于SMA领域，通过强制表达8个已定义的转录因子，获得了患者特异性的诱导运动神经元，这些神经元直接从成纤维细胞转化而来。感染的成纤维细胞开始以偶极方式生长，细胞核逐渐增大。第23天出现典型的Tuj1阳性神经元。35d后，观察到诱导后的神经元有多个突起，这些神经元还表达Tuj1、HB9、Isl1和ChAT的特征。SMA组和对照组的转换效率分别约为5.8%和5.5%。此外，与对照神经元相比，SMA诱导的神经元突起生长速度显著降低。60d后，SMA诱导的神经元也表现出神经元变性的倾向，并观察到明显的突起断裂。通过直接对成纤维细胞重新编程，我们建立了一个无饲养层的转换系统，以获得SMA患者特有的诱导运动神经元，该神经元在体外部分模拟了SMA的表型。
Spinal muscular atrophy (SMA) is a lethal autosomal recessive neurological disease characterized by selective degeneration of motor neurons in the spinal cord. In recent years, the development of cellular reprogramming technology has provided an alternative and effective method for obtaining patient-specific neurons in vitro. In the present study, we applied this technology to the field of SMA to acquire patient-specific induced motor neurons that were directly converted from fibroblasts via the forced expression of 8 defined transcription factors. The infected fibroblasts began to grow in a dipolar manner, and the nuclei gradually enlarged. Typical Tuj1-positive neurons were generated at day 23. After day 35, induced neurons with multiple neurites were observed, and these neurons also expressed the hallmarks of Tuj1, HB9, ISL1 and CHAT. The conversion efficiencies were approximately 5.8% and 5.5% in the SMA and control groups, respectively. Additionally, the SMA-induced neurons exhibited a significantly reduced neurite outgrowth rate compared with the control neurons. After day 60, the SMA-induced neurons also exhibited a liability of neuronal degeneration and remarkable fracturing of the neurites was observed. By directly reprogramming fibroblasts, we established a feeder-free conversion system to acquire SMA patient-specific induced motor neurons that partially modeled the phenotype of SMA in vitro.