Hansen Susanne K, Stummann Tina C, Borland Helena, Hasholt Lis F, Tümer Zeynep, Nielsen Jørgen E, Rasmussen Mikkel A, Nielsen Troels T, Daechsel Justus C A, Fog Karina, Hyttel Poul
Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, Groennegårdsvej 7, 1870 Frb C, Denmark; H. Lundbeck A/S, Ottiliavej 9, Valby 2500, Denmark.
H. Lundbeck A/S, Ottiliavej 9, Valby 2500, Denmark.
Stem Cell Res. 2016 Sep;17(2):306-317. doi: 10.1016/j.scr.2016.07.004. Epub 2016 Aug 16.
The neurodegenerative disease spinocerebellar ataxia type 3 (SCA3) is caused by a CAG-repeat expansion in the ATXN3 gene. In this study, induced pluripotent stem cell (iPSC) lines were established from two SCA3 patients. Dermal fibroblasts were reprogrammed using an integration-free method and the resulting SCA3 iPSCs were differentiated into neurons. These neuronal lines harbored the disease causing mutation, expressed comparable levels of several neuronal markers and responded to the neurotransmitters, glutamate/glycine, GABA and acetylcholine. Additionally, all neuronal cultures formed networks displaying synchronized spontaneous calcium oscillations within 28days of maturation, and expressed the mature neuronal markers NeuN and Synapsin 1 implying a relatively advanced state of maturity, although not comparable to that of the adult human brain. Interestingly, we were not able to recapitulate the glutamate-induced ataxin-3 aggregation shown in a previously published iPSC-derived SCA3 model. In conclusion, we have generated a panel of SCA3 patient iPSCs and a robust protocol to derive neurons of relatively advanced maturity, which could potentially be valuable for the study of SCA3 disease mechanisms.
神经退行性疾病3型脊髓小脑共济失调（SCA3）由ATXN3基因中的CAG重复扩增引起。在本研究中，从两名SCA3患者建立了诱导多能干细胞（iPSC）系。使用无整合方法将皮肤成纤维细胞重编程，所得的SCA3 iPSC分化为神经元。这些神经元系携带致病突变，表达几种神经元标志物的水平相当，并对神经递质谷氨酸/甘氨酸、GABA和乙酰胆碱有反应。此外，所有神经元培养物在成熟的28天内形成了显示同步自发钙振荡的网络，并表达了成熟神经元标志物NeuN和突触素1，这意味着处于相对成熟的状态，尽管与成人人类大脑的成熟状态不可比。有趣的是，我们无法重现先前发表的iPSC衍生的SCA3模型中所示的谷氨酸诱导的ataxin-3聚集。总之，我们生成了一组SCA3患者iPSC和一个强大的方案来衍生相对成熟的神经元，这可能对SCA3疾病机制的研究有潜在价值。
