Carmona Vitor, Cunha-Santos Janete, Onofre Isabel, Simões Ana Teresa, Vijayakumar Udaya, Davidson Beverly L, Pereira de Almeida Luís
CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Rua Larga, Coimbra 3004-504, Portugal; Faculty of Pharmacy, University of Coimbra, Coimbra 3000-548, Portugal.
CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Rua Larga, Coimbra 3004-504, Portugal.
Mol Ther. 2017 Apr 5;25(4):1038-1055. doi: 10.1016/j.ymthe.2017.01.021. Epub 2017 Feb 22.
Machado-Joseph disease (MJD) is a genetic neurodegenerative disease caused by an expanded polyglutamine tract within the protein ataxin-3 (ATXN3). Despite current efforts, MJD's mechanism of pathogenesis remains unclear and no disease-modifying treatment is available. Therefore, in this study, we investigated (1) the role of the 3' UTR of ATXN3, a putative microRNA (miRNA) target, (2) whether miRNA biogenesis and machinery are dysfunctional in MJD, and (3) which specific miRNAs target ATXN3-3' UTR and whether they can alleviate MJD neuropathology in vivo. Our results demonstrate that endogenous miRNAs, by targeting sequences in the 3' UTR, robustly reduce ATXN3 expression and aggregation in vitro and neurodegeneration and neuroinflammation in vivo. Importantly, we found an abnormal MJD-associated downregulation of genes involved in miRNA biogenesis and silencing activity. Finally, we identified three miRNAs-mir-9, mir-181a, and mir-494-that interact with the ATXN3-3' UTR and whose expression is dysregulated in human MJD neurons and in other MJD cell and animal models. Furthermore, overexpression of these miRNAs in mice resulted in reduction of mutATXN3 levels, aggregate counts, and neuronal dysfunction. Altogether, these findings indicate that endogenous miRNAs and the 3' UTR of ATXN3 play a crucial role in MJD pathogenesis and provide a promising opportunity for MJD treatment.
马查多-约瑟夫病(MJD)是一种由ataxin-3(ATXN3)蛋白内多聚谷氨酰胺序列扩增引起的遗传性神经退行性疾病。尽管目前已做出诸多努力,但MJD的发病机制仍不清楚,且尚无改善病情的治疗方法。因此,在本研究中,我们调查了:(1)ATXN3的3'非翻译区(3'UTR)作为假定的微小RNA(miRNA)靶点的作用;(2)MJD中miRNA的生物合成和作用机制是否存在功能障碍;(3)哪些特定的miRNA靶向ATXN3的3'UTR,以及它们是否能在体内减轻MJD的神经病理学变化。我们的结果表明,内源性miRNA通过靶向3'UTR中的序列,在体外能显著降低ATXN3的表达和聚集,在体内能减轻神经退行性变和神经炎症。重要的是,我们发现与miRNA生物合成和沉默活性相关的基因在MJD中出现异常下调。最后,我们鉴定出三种miRNA——mir-9、mir-181a和mir-494,它们与ATXN3的3'UTR相互作用,且在人类MJD神经元以及其他MJD细胞和动物模型中的表达失调。此外,在小鼠中过表达这些miRNA可导致突变型ATXN3水平降低、聚集体数量减少以及神经元功能障碍。总之,这些发现表明内源性miRNA和ATXN3的3'UTR在MJD发病机制中起关键作用,并为MJD的治疗提供了一个有前景的机会。