Amado Defne A, Robbins Ashley B, Whiteman Katherine R, Smith Alicia R, Chillon Guillem, Chen Yonghong, Fuller Joshua A, Patty Nicholas A, Izda Aleksandar, Cheng Congsheng, Nelson Shareen, Dichter Abigail I, Mazzoni Esteban O, Monteys Alex Mas, Davidson Beverly L
Department of Neurology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Raymond G. Perelman Center for Cellular and Molecular Therapeutics, Children's Hospital of Philadelphia, Philadelphia, PA, USA.
Nat Commun. 2025 Jun 25;16(1):5334. doi: 10.1038/s41467-025-60497-8.
Amyotrophic lateral sclerosis (ALS) involves motor neuron death due to mislocalized TDP-43. Pathologic TDP-43 associates with stress granules (SGs), and lowering the SG-associated protein ataxin-2 (ATXN2) using Atxn2-targeting antisense oligonucleotides prolongs survival in TAR4/4 sporadic ALS mice but failed in clinical trials likely due to poor target engagement. Here we show that an AAV with potent motor neuron transduction delivering Atxn2-targeting miRNAs reduces Atxn2 throughout the central nervous system at doses 40x lower than published work. In TAR4/4 mice, miAtxn2 increased survival (50%) and strength, and reduced motor neuron death, inflammation, and phosphorylated TDP-43. TAR4/4 transcriptomic dysregulation recapitulated ALS gene signatures that were rescued by miAtxn2, identifying potential therapeutic mechanisms and biomarkers. In slow progressing hemizygous mice, miAtxn2 slowed disease progression, and in ALS patient-derived lower motor neurons, our AAV vector transduced >95% of cells and potently reduced ATXN2 at MOI 4 logs lower than previously reported. These data support AAV-RNAi targeting ATXN2 as a translatable therapy for sporadic ALS.
肌萎缩侧索硬化症（ALS）涉及因TDP-43定位错误导致的运动神经元死亡。病理性TDP-43与应激颗粒（SGs）相关，使用靶向Atxn2的反义寡核苷酸降低与SG相关的蛋白质ataxin-2（ATXN2）可延长TAR4/4散发性ALS小鼠的生存期，但在临床试验中失败，可能是由于靶点参与度不佳。在此我们表明，一种具有高效运动神经元转导能力的腺相关病毒（AAV）递送靶向Atxn2的微小RNA（miRNA），能以比已发表研究低40倍的剂量在整个中枢神经系统中降低Atxn2水平。在TAR4/4小鼠中，miAtxn2提高了生存率（50%）和力量，减少了运动神经元死亡、炎症以及磷酸化TDP-43。TAR4/4的转录组失调重现了ALS基因特征，而miAtxn2可使其得到挽救，从而确定了潜在的治疗机制和生物标志物。在进展缓慢的半合子小鼠中，miAtxn2减缓了疾病进展，在ALS患者来源的下运动神经元中，我们的AAV载体在比先前报道低4个对数的感染复数（MOI）下转导了>95%的细胞，并有效降低了ATXN2。这些数据支持将靶向ATXN2的AAV-RNAi作为散发性ALS的一种可转化治疗方法。
