Department of Human Genetics, Leiden University Medical Center, Leiden, Zuid-Holland, The Netherlands.
Department of Neurology, Leiden University Medical Center, Leiden, Zuid-Holland, The Netherlands.
Mov Disord. 2023 Aug;38(8):1428-1442. doi: 10.1002/mds.29446. Epub 2023 Jun 6.
Spinocerebellar ataxia type 1 (SCA1) is a neurodegenerative disease caused by a polyglutamine expansion in the ataxin-1 protein resulting in neuropathology including mutant ataxin-1 protein aggregation, aberrant neurodevelopment, and mitochondrial dysfunction.
Identify SCA1-relevant phenotypes in patient-specific fibroblasts and SCA1 induced pluripotent stem cells (iPSCs) neuronal cultures.
SCA1 iPSCs were generated and differentiated into neuronal cultures. Protein aggregation and neuronal morphology were evaluated using fluorescent microscopy. Mitochondrial respiration was measured using the Seahorse Analyzer. The multi-electrode array (MEA) was used to identify network activity. Finally, gene expression changes were studied using RNA-seq to identify disease-specific mechanisms.
Bioenergetics deficits in patient-derived fibroblasts and SCA1 neuronal cultures showed altered oxygen consumption rate, suggesting involvement of mitochondrial dysfunction in SCA1. In SCA1 hiPSC-derived neuronal cells, nuclear and cytoplasmic aggregates were identified similar in localization as aggregates in SCA1 postmortem brain tissue. SCA1 hiPSC-derived neuronal cells showed reduced dendrite length and number of branching points while MEA recordings identified delayed development in network activity in SCA1 hiPSC-derived neuronal cells. Transcriptome analysis identified 1050 differentially expressed genes in SCA1 hiPSC-derived neuronal cells associated with synapse organization and neuron projection guidance, where a subgroup of 151 genes was highly associated with SCA1 phenotypes and linked to SCA1 relevant signaling pathways.
Patient-derived cells recapitulate key pathological features of SCA1 pathogenesis providing a valuable tool for the identification of novel disease-specific processes. This model can be used for high throughput screenings to identify compounds, which may prevent or rescue neurodegeneration in this devastating disease. © 2023 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.
脊髓小脑共济失调 1 型(SCA1)是一种神经退行性疾病,由ataxin-1 蛋白中的多聚谷氨酰胺扩展引起,导致神经病理学改变,包括突变型 ataxin-1 蛋白聚集、异常神经发育和线粒体功能障碍。
在患者特异性成纤维细胞和 SCA1 诱导多能干细胞(iPSC)神经元培养物中鉴定与 SCA1 相关的表型。
生成 SCA1 iPSC 并分化为神经元培养物。使用荧光显微镜评估蛋白聚集和神经元形态。使用 Seahorse 分析仪测量线粒体呼吸。使用多电极阵列(MEA)鉴定网络活动。最后,使用 RNA-seq 研究基因表达变化,以鉴定疾病特异性机制。
患者来源的成纤维细胞和 SCA1 神经元培养物的生物能缺陷显示耗氧率改变,提示线粒体功能障碍参与 SCA1。在 SCA1 hiPSC 衍生的神经元细胞中,鉴定到核内和细胞质聚集,其定位与 SCA1 尸检脑组织中的聚集相似。SCA1 hiPSC 衍生的神经元细胞表现出树突长度和分支点数量减少,而 MEA 记录表明 SCA1 hiPSC 衍生的神经元细胞网络活动发育延迟。转录组分析鉴定出 SCA1 hiPSC 衍生的神经元细胞中 1050 个差异表达基因与突触组织和神经元投射导向有关,其中一组 151 个基因与 SCA1 表型高度相关,并与 SCA1 相关信号通路有关。
患者来源的细胞再现了 SCA1 发病机制的关键病理特征,为鉴定新的疾病特异性过程提供了有价值的工具。该模型可用于高通量筛选,以鉴定可能预防或挽救这种毁灭性疾病中神经退行性变的化合物。© 2023 作者。运动障碍由 Wiley 期刊出版社代表国际帕金森和运动障碍学会出版。