Identification of therapeutic targets for ALS by screening for TDP-43 and FUS associated molecules

通过引入编码FUS基因shRNA的慢病毒，使小鼠原代皮质神经元被FUS基因沉默。接下来，我们使用外显子敏感微阵列建立了FUS调节的初级神经元转录组谱，并确定FUS调节Mapt外显子10的选择性剪接。这种选择性剪接事件导致三重复（RD3）和四重复（RD4） Tau亚型的产生，当FUS在神经元中沉默时，RD4/RD3的比例增加。在fus沉默的原代神经元中观察到神经突的形态异常，并通过RD4的沉默修复。这些发现表明，RD4可能是ALS治疗的一个治疗靶点。

Mouse primary cortical neurons were silenced with FUS by introducing lentivirus encoding shRNA against FUS gene. We next established the FUS-regulated transcriptome profiles of primary neurons using exon-sensitive microarrays and identified that FUS regulated the alternative splicing of Mapt exon10. This alternative splicing event results in three-repeats (RD3) and four-repeats (RD4) Tau isoform production and the ratio of RD4/RD3 was increased when FUS was silenced in neurons. Morphological abnormality of neurite was observed in FUS-silenced primary neurons and it was rescued bysilencing of RD4. These findings suggest that RD4 could be a therapeutic target for ALS treatment.