Progress in Neurobiology ( IF 6.7 ) Pub Date : 2020-04-24 , DOI: 10.1016/j.pneurobio.2020.101803 Alessia Loffreda 1 , Monica Nizzardo 2 , Alessandro Arosio 3 , Marc-David Ruepp 4 , Raffaele A Calogero 5 , Stefano Volinia 6 , Marco Galasso 6 , Caterina Bendotti 7 , Carlo Ferrarese 8 , Christian Lunetta 9 , Mafalda Rizzuti 2 , Antonella E Ronchi 1 , Oliver Mühlemann 4 , Lucio Tremolizzo 8 , Stefania Corti 10 , Silvia M L Barabino 1
Amyotrophic lateral sclerosis (ALS) is a relentless and fatal neurological disease characterized by the selective degeneration of motor neurons. No effective therapy is available for this disease. Several lines of evidence indicate that alteration of RNA metabolism, including microRNA (miRNA) processing, is a relevant pathogenetic factor and a possible therapeutic target for ALS.
Here, we showed that the abundance of components in the miRNA processing machinery is altered in a SOD1-linked cellular model, suggesting consequent dysregulation of miRNA biogenesis. Indeed, high-throughput sequencing of the small RNA fraction showed that among the altered miRNAs, miR-129-5p was increased in different models of SOD1-linked ALS and in peripheral blood cells of sporadic ALS patients. We demonstrated that miR-129-5p upregulation causes the downregulation of one of its targets: the RNA-binding protein ELAVL4/HuD. ELAVL4/HuD is predominantly expressed in neurons, where it controls several key neuronal mRNAs. Overexpression of pre-miR-129-1 inhibited neurite outgrowth and differentiation via HuD silencing in vitro, while its inhibition with an antagomir rescued the phenotype.
Remarkably, we showed that administration of an antisense oligonucleotide (ASO) inhibitor of miR-129-5p to an ALS animal model, SOD1 (G93A) mice, result in a significant increase in survival and improved the neuromuscular phenotype in treated mice. These results identify miR-129-5p as a therapeutic target that is amenable to ASO modulation for the treatment of ALS patients.
中文翻译:
miR-129-5p:肌萎缩性侧索硬化症的关键因素和治疗目标。
肌萎缩性侧索硬化症(ALS)是一种以运动神经元选择性变性为特征的无情和致命性神经疾病。对于这种疾病没有有效的治疗方法。几条证据表明,RNA代谢的改变(包括microRNA(miRNA)加工)是相关的致病因素,也是ALS的可能治疗靶标。
在这里,我们表明,在与SOD1相连的细胞模型中,miRNA处理机制中的组分丰度发生了变化,表明随后的miRNA生物发生失调。的确,对小分子RNA片段的高通量测序表明,在改变的miRNA中,在不同模型的SOD1连接的ALS和散发性ALS患者的外周血细胞中,miR-129-5p升高。我们证明了miR-129-5p的上调会导致其靶标之一的下调:RNA结合蛋白ELAVL4 / HuD。ELAVL4 / HuD主要在神经元中表达,它控制着几个关键的神经元mRNA。体外通过HuD沉默,pre-miR-129-1的过表达抑制了神经突生长和分化。,而用antagomir抑制则拯救了该表型。
值得注意的是,我们显示了将miR-129-5p的反义寡核苷酸(ASO)抑制剂给予ALS动物模型SOD1(G93A)小鼠,可显着提高存活率并改善治疗小鼠的神经肌肉表型。这些结果确定了miR-129-5p是适合于ASO调节以治疗ALS患者的治疗靶标。