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

通过引入编码针对FUS基因的shRNA的慢病毒，用FUS沉默小鼠原代皮层神经元。接下来，我们使用外显子敏感的微阵列建立了FUS调节的原代神经元转录组谱，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（RD 3）和四重复（RD 4）Tau同种型的产生，并且当神经元中FUS沉默时，RD 4/RD 3的比率增加。在FUS沉默的原代神经元中观察到突起的形态学异常，并通过沉默RD 4来挽救。这些发现表明RD 4可能是ALS治疗的治疗靶点。

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.