肌萎缩性侧索硬化症药物发现通过高通量表型筛选使用iPSC-Derived人类运动神经元

肌萎缩性脊髓侧索硬化(ALS)是一种神经退行性疾病主要影响运动神经元。不幸的是,只有两种药物治疗条件,批准这两种提高病人生存超过几个月。这个发人深省的现实表明迫切需要新的ALS治疗的发展,一直饱受高失败率的候选药物在临床试验。这么高的失败率表明临床筛查策略需要重新评估。ALS患者的疾病的标志之一是低表达的异常神经丝轻链(NFL)在运动神经元。此外,联盟的恢复到正常水平防止表型标志ALS神经元的变化。因此,我们希望建立一个临床相关的筛选平台,确定化合物NFL的表情回到正常水平ALS病人运动神经元。在BrainXell,我们建立了新技术快速区分ALS患者诱导多能干细胞(万能)大量的神经元。然后我们使用基因组编辑技术与nanoluciferase开保险丝NFL (NLuc)记者,从而使高通量筛选(高温超导)系统监控NFL的表达水平72 h后每个化合物接触。分析适应满足高温超导的要求,包括:大型批量大小,1536 -格式,最小的井间变异,短期文化、电镀自动分发器,低试剂卷。 Applying a quantitative HTS approach, we screened the LOPAC, NPC, and MIPE libraries (>6,000 compounds) in a dose dependent manner. Compounds that increase NFL expression by >30% (to approximately normal levels) were considered hits. From these screens we identified 80 hit compounds that are currently going through secondary validation. Preliminary data look promising. For example, two of these hits restore normal expression of NFL with no observed toxicity.