Laboratory of Cellular and Molecular Neurobiology - Stem Cells, Department of Neurobiology, Hellenic Pasteur Institute, 127 Vassilissis Sofias Avenue, 11521, Athens, Greece.
Faculty of Nursing, University of Athens, 11527, Athens, Greece.
Neurochem Res. 2019 Jun;44(6):1475-1493. doi: 10.1007/s11064-019-02781-w. Epub 2019 Apr 15.
Parkinson's disease (PD) is the second most common neurodegenerative disorder. We have previously developed a disease-in-a-dish model for familial PD using induced pluripotent stem cells (iPSCs) from two patients carrying the p.A53T α-synuclein (αSyn) mutation. By directed differentiation, we generated a model that displays disease-relevant phenotypes, including protein aggregation, compromised neurite outgrowth, axonal neuropathology and synaptic defects. Here we investigated the in vivo phenotypes of iPSCs, derived from one patient, after transplantation in a lesion mouse model established by unilateral intrastriatal 6-hydroxydopamine injection in the immunosuppressed NOD/SCID strain. Immunohistochemistry revealed that despite the disease-related characteristics that mutant cells displayed when maintained up to 70 days in vitro, they could survive and differentiate in vivo over a 12-week period. However, some differences were noted between patient-derived and control grafts, including a significant rise in αSyn immunoreactivity that might signal a first step towards pathology. Moreover, control-derived grafts appeared to integrate better than PD grafts within the host tissue extending projections that formed more contacts with host striatal neurons. Our data suggest that the distinct disease-related characteristics which p.A53T cells develop in vitro, may be attenuated or take longer to emerge in vivo after transplantation within the mouse brain. Further analysis of the phenotypes that patient cells acquire over longer periods of time as well as the use of multiple iPSC clones from different patients should extend our current proof-of-concept study and provide additional evidence for in vivo disease modeling.
帕金森病（PD）是第二常见的神经退行性疾病。我们之前使用携带 p.A53T α-突触核蛋白（αSyn）突变的两位患者的诱导多能干细胞（iPSC）开发了一种家族性 PD 的疾病模型。通过定向分化，我们生成了一种显示疾病相关表型的模型，包括蛋白聚集、神经突生长受损、轴突神经病理学和突触缺陷。在这里，我们研究了源自一位患者的 iPSC 的体内表型，这些 iPSC 是在免疫抑制的 NOD/SCID 品系通过单侧纹状体 6-羟多巴胺注射建立的病变小鼠模型中进行移植后获得的。免疫组织化学显示，尽管突变细胞在体外培养至 70 天时表现出与疾病相关的特征，但它们可以在体内存活和分化超过 12 周。然而，在患者来源的和对照移植物之间观察到一些差异，包括αSyn 免疫反应性的显著增加，这可能是病理学的第一步。此外，与 PD 移植物相比，对照来源的移植物似乎在宿主组织内更好地整合，延伸的突起与宿主纹状体神经元形成更多的接触。我们的数据表明，p.A53T 细胞在体外发展的独特与疾病相关的特征，在移植到小鼠大脑后，可能会减弱或需要更长时间才能在体内出现。进一步分析患者细胞在更长时间内获得的表型以及使用来自不同患者的多个 iPSC 克隆，应该可以扩展我们当前的概念验证研究，并为体内疾病建模提供更多证据。
