Loss of spastin function results in disease-specific axonal defects in human pluripotent stem cell-based models of hereditary spastic paraplegia.
Loss of spastin function results in disease-specific axonal defects in human pluripotent stem cell-based models of hereditary spastic paraplegia.
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DOI：
10.1002/stem.1569
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发表时间：
2014-02
期刊：
影响因子：
5.2
通讯作者：
Li, Xue-Jun
中科院分区：
文献类型：
--
作者：
Denton, Kyle R.;Lei, Ling;Grenier, Jeremy;Rodionov, Vladimir;Blackstone, Craig;Li, Xue-Jun
关键词：
遗传性痉挛性截瘫（HSP）的人类神经元模型概括了疾病特异性轴突病理学，这是理解患者中某些轴突退化的原因和开发治疗方法的关键。SPG4是HSP最常见的形式，由SPAST基因中的常染色体显性突变引起，该基因编码微管切断ATP酶痉挛蛋白。在这里，我们通过从SPG4患者中建立诱导多能干细胞（iPSC）并将这些细胞分化为端脑神经元来产生SPG4的人类神经元模型。SPG4神经元显示轴突sweep显著增加，其对线粒体和tau蛋白染色强烈，表明轴突运输货物的积累。此外，SPG4神经元中线粒体转运减少，揭示了这些患者iPSC衍生的神经元重现了疾病特异性轴突表型。有趣的是，SPG4神经元中spastin蛋白水平显著降低，支持单倍不足机制。此外，源自spastin敲低的人胚胎干细胞（hESC）的皮质神经元表现出类似的轴突缺陷，证实了轴突缺陷可以由spastin功能丧失引起。这些spastin敲低的hESC作为研究HSP的另一个模型。最后，SPG 4神经元中稳定的乙酰化微管蛋白水平显着增加。长春碱是一种微管去稳定化药物，在来源于SPG4 iPSC和spastin敲低的hESC的神经元中挽救了这种轴突肿胀表型。因此，本研究成功建立了SPG4神经元模型，为进一步研究HSP的致病细胞机制和筛选拯救HSP轴突变性的药物提供了实验依据。
Human neuronal models of hereditary spastic paraplegias (HSP) that recapitulate disease-specific axonal pathology hold the key to understanding why certain axons degenerate in patients and to developing therapies. SPG4, the most common form of HSP, is caused by autosomal dominant mutations in the SPAST gene, which encodes the microtubule-severing ATPase spastin. Here, we have generated a human neuronal model of SPG4 by establishing induced pluripotent stem cells (iPSCs) from an SPG4 patient and differentiating these cells into telencephalic glutamatergic neurons. The SPG4 neurons displayed a significant increase in axonal swellings, which stained strongly for mitochondria and tau, indicating the accumulation of axonal transport cargoes. In addition, mitochondrial transport was decreased in SPG4 neurons, revealing that these patient iPSC-derived neurons recapitulate disease-specific axonal phenotypes. Interestingly, spastin protein levels were significantly decreased in SPG4 neurons, supporting a haploinsufficiency mechanism. Furthermore, cortical neurons derived from spastin-knockdown human embryonic stem cells (hESCs) exhibited similar axonal swellings, confirming that the axonal defects can be caused by loss of spastin function. These spastin-knockdown hESCs serve as an additional model for studying HSP. Finally, levels of stabilized acetylated-tubulin were significantly increased in SPG4 neurons. Vinblastine, a microtubule-destabilizing drug, rescued this axonal swelling phenotype in neurons derived from both SPG4 iPSCs and spastin-knockdown hESCs. Thus, this study demonstrates the successful establishment of human pluripotent stem cell-based neuronal models of SPG4, which will be valuable for dissecting the pathogenic cellular mechanisms and screening compounds to rescue the axonal degeneration in HSP.
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影响因子：
4.3
作者：
Abrahamsen G;Fan Y;Matigian N;Wali G;Bellette B;Sutharsan R;Raju J;Wood SA;Veivers D;Sue CM;Mackay-Sim A
通讯作者：
Mackay-Sim A
影响因子：
7.8
作者：
Baas, P W;Black, M M
通讯作者：
Black, M M
DOI：
10.1083/jcb.200409058
发表时间：
2005-02-14
期刊：
The Journal of cell biology
影响因子：
--
作者：
Evans KJ;Gomes ER;Reisenweber SM;Gundersen GG;Lauring BP
通讯作者：
Lauring BP
影响因子：
30.8
作者：
Hazan, J;Fonknechten, N;Weissenbach, J
通讯作者：
Weissenbach, J
影响因子：
16.2
作者：
Arlotta, P;Molyneaux, BJ;Macklis, JD
通讯作者：
Macklis, JD