Amyotrophic lateral sclerosis (ALS) is currently an incurable disease without highly effective pharmacological treatments. The peptide drug GM604 (GM6 or Alirinetide) was developed as a candidate ALS therapy, which has demonstrated safety and good drug-like properties with a favorable pharmacokinetic profile. GM6 is hypothesized to bolster neuron survival through the multi-target regulation of developmental pathways, but mechanisms of action are not fully understood. This study used RNA-seq to evaluate transcriptome responses in SH-SY5Y neuroblastoma cells following GM6 treatment (6, 24 and 48 h). We identified 2867 protein-coding genes with expression significantly altered by GM6 (FDR < 0.10). Early (6 h) responses included up-regulation of Notch and hedgehog signaling components, with increased expression of developmental genes mediating neurogenesis and axon growth. Prolonged GM6 treatment (24 and 48 h) altered the expression of genes contributing to cell adhesion and the extracellular matrix. GM6 further down-regulated the expression of genes associated with mitochondria, inflammatory responses, mRNA processing and chromatin organization. GM6-increased genes were located near GC-rich motifs interacting with C2H2 zinc finger transcription factors, whereas GM6-decreased genes were located near AT-rich motifs associated with helix-turn-helix homeodomain factors. Such motifs interacted with a diverse network of transcription factors encoded by GM6-regulated genes (STAT3, HOXD11, HES7, GLI1). We identified 77 ALS-associated genes with expression significantly altered by GM6 treatment (FDR < 0.10), which were known to function in neurogenesis, axon guidance and the intrinsic apoptosis pathway. Our findings support the hypothesis that GM6 acts through developmental-stage pathways to influence neuron survival. Gene expression responses were consistent with neurotrophic effects, ECM modulation, and activation of the Notch and hedgehog neurodevelopmental pathways. This multifaceted mechanism of action is unique among existing ALS drug candidates and may be applicable to multiple neurodegenerative diseases.

中文翻译：

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GM604 调节发育神经发生途径和与肌萎缩侧索硬化症相关的基因的表达。

肌萎缩侧索硬化症（ALS）目前是一种无法治愈的疾病，没有高效的药物治疗。肽药物 GM604（GM6 或 Alirinetide）被开发为候选 ALS 疗法，已证明其安全性和良好的药物样特性以及良好的药代动力学特征。GM6 被假设通过发育途径的多靶点调节来支持神经元存活，但作用机制尚不完全清楚。本研究使用 RNA-seq 评估 GM6 处理（6、24 和 48 小时）后 SH-SY5Y 神经母细胞瘤细胞中的转录组反应。我们鉴定了 2867 个蛋白质编码基因，其表达被 GM6 显着改变（FDR < 0.10）。早期（6小时）反应包括Notch和hedgehog信号成分的上调，随着介导神经发生和轴突生长的发育基因表达增加。延长 GM6 处理（24 和 48 小时）改变了有助于细胞粘附和细胞外基质的基因的表达。GM6 进一步下调与线粒体、炎症反应、mRNA 加工和染色质组织相关的基因的表达。GM6 增加的基因位于与 C2H2 锌指转录因子相互作用的富含 GC 的基序附近，而 GM6 减少的基因位于与螺旋-转角-螺旋同源域因子相关的富含 AT 的基序附近。这些基序与由 GM6 调节基因（STAT3、HOXD11、HES7、GLI1）编码的多种转录因子网络相互作用。我们鉴定了 77 个 ALS 相关基因，其表达因 GM6 处理而显着改变（FDR < 0.10），已知它们在神经发生、轴突引导和内在细胞凋亡途径中起作用。我们的研究结果支持 GM6 通过发育阶段途径影响神经元存活的假设。基因表达反应与神经营养作用、ECM 调节以及 Notch 和刺猬神经发育通路的激活一致。这种多方面的作用机制在现有的 ALS 候选药物中是独一无二的，可能适用于多种神经退行性疾病。