Bauer Peter O, Dunmore Judith H, Sasaguri Hiroki, Matoska Vaclav
Department of Neuroscience, Mayo Clinic, Jacksonville, FL, United States.
Bioinova, Ltd., Prague, Czechia.
Front Neurosci. 2019 Sep 6;13:935. doi: 10.3389/fnins.2019.00935. eCollection 2019.
Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are incurable neurodegenerative conditions. A non-coding hexanucleotide (GGGGCC) repeat expansion in the gene is the most common genetic cause of ALS/FTD. We present a cellular model of c9ALS/FTD where induced neurons (iNeurons) are generated within 2 weeks by direct conversion of patients' dermal fibroblasts through down-regulation of polypyrimidine-tract-binding protein 1 (PTB1). While sense (S) and anti-sense (AS) intranuclear RNA foci were observed in both fibroblasts and iNeurons, the accumulation of (S) and (AS) repeat-associated non-ATG translation (RANT) products were detected only in iNeurons. Importantly, anti-sense oligonucleotides (ASOs) against the (S) repeat transcript lead to decreased (S) RNA foci staining and a reduction of the corresponding RANT products without affecting its (AS) counterparts. ASOs treatment also rescued the cell viability upon stressful stimulus. The results indicate that iNeurons is an advantageous model that not only recapitulates c9ALS/FTD hallmark features but can also help uncover promising therapeutics.
肌萎缩侧索硬化症（ALS）和额颞叶痴呆（FTD）是无法治愈的神经退行性疾病。该基因中的非编码六核苷酸（GGGGCC）重复扩增是ALS/FTD最常见的遗传病因。我们提出了一种c9ALS/FTD细胞模型，其中通过下调聚嘧啶序列结合蛋白1（PTB1），在2周内将患者的皮肤成纤维细胞直接转化生成诱导神经元（iNeurons）。虽然在成纤维细胞和iNeurons中均观察到正义（S）和反义（AS）核内RNA病灶，但仅在iNeurons中检测到（S）和（AS）重复相关的非ATG翻译（RANT）产物的积累。重要的是，针对（S）重复转录本的反义寡核苷酸（ASO）可导致（S）RNA病灶染色减少以及相应RANT产物的减少，而不影响其（AS）对应物。ASO处理还可在应激刺激下挽救细胞活力。结果表明，iNeurons是一种优势模型，不仅可重现c9ALS/FTD的标志性特征，还可帮助发现有前景的治疗方法。
