CNC-Center for Neurosciences and Cell Biology, University of Coimbra, Coimbra, Portugal.
CIBB-Center for Innovative Biomedicine and Biotechnology, University of Coimbra, Coimbra, Portugal.
Sci Rep. 2022 Jul 22;12(1):12513. doi: 10.1038/s41598-022-16369-y.
The establishment of robust human brain organoids to model cerebellar diseases is essential to study new therapeutic strategies for cerebellum-associated disorders. Machado-Joseph disease (MJD) is a cerebellar hereditary neurodegenerative disease, without therapeutic options able to prevent the disease progression. In the present work, control and MJD induced-pluripotent stem cells were used to establish human brain organoids. These organoids were characterized regarding brain development, cell type composition, and MJD-associated neuropathology markers, to evaluate their value for cerebellar diseases modeling. Our data indicate that the organoids recapitulated, to some extent, aspects of brain development, such as astroglia emerging after neurons and the presence of ventricular-like zones surrounded by glia and neurons that are found only in primate brains. Moreover, the brain organoids presented markers of neural progenitors proliferation, neuronal differentiation, inhibitory and excitatory synapses, and firing neurons. The established brain organoids also exhibited markers of cerebellar neurons progenitors and mature cerebellar neurons. Finally, MJD brain organoids showed higher ventricular-like zone numbers, an indication of lower maturation, and an increased number of ataxin-3-positive aggregates, compared with control organoids. Altogether, our data indicate that the established organoids recapitulate important characteristics of human brain development and exhibit cerebellar features, constituting a resourceful tool for testing therapeutic approaches for cerebellar diseases.
建立稳健的人类大脑类器官来模拟小脑疾病对于研究与小脑相关疾病的新治疗策略至关重要。马查多-约瑟夫病（MJD）是一种小脑遗传性神经退行性疾病，目前尚无能够预防疾病进展的治疗方法。在本工作中，使用对照和 MJD 诱导的多能干细胞建立了人类大脑类器官。这些类器官在脑发育、细胞类型组成和 MJD 相关神经病理学标志物方面进行了特征分析，以评估它们在小脑疾病建模中的价值。我们的数据表明，这些类器官在一定程度上再现了脑发育的某些方面，例如神经元之后出现的星形胶质细胞，以及仅在灵长类动物脑中存在的被胶质细胞和神经元包围的脑室样区域。此外，脑类器官还表现出神经祖细胞增殖、神经元分化、抑制性和兴奋性突触以及放电神经元的标志物。建立的大脑类器官还表现出小脑神经元祖细胞和成熟小脑神经元的标志物。最后，与对照类器官相比，MJD 大脑类器官的脑室样区域数量更多，这表明成熟度较低，并且聚集的抗肌萎缩蛋白-3阳性颗粒数量增加。总之，我们的数据表明，所建立的类器官再现了人类大脑发育的重要特征，并表现出小脑特征，为测试小脑疾病的治疗方法提供了有价值的工具。
