Didiot Marie-Cécile, Hall Lauren M, Coles Andrew H, Haraszti Reka A, Godinho Bruno Mdc, Chase Kathryn, Sapp Ellen, Ly Socheata, Alterman Julia F, Hassler Matthew R, Echeverria Dimas, Raj Lakshmi, Morrissey David V, DiFiglia Marian, Aronin Neil, Khvorova Anastasia
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Mol Ther. 2016 Oct;24(10):1836-1847. doi: 10.1038/mt.2016.126. Epub 2016 Jun 27.
Delivery represents a significant barrier to the clinical advancement of oligonucleotide therapeutics for the treatment of neurological disorders, such as Huntington's disease. Small, endogenous vesicles known as exosomes have the potential to act as oligonucleotide delivery vehicles, but robust and scalable methods for loading RNA therapeutic cargo into exosomes are lacking. Here, we show that hydrophobically modified small interfering RNAs (hsiRNAs) efficiently load into exosomes upon co-incubation, without altering vesicle size distribution or integrity. Exosomes loaded with hsiRNAs targeting Huntingtin mRNA were efficiently internalized by mouse primary cortical neurons and promoted dose-dependent silencing of Huntingtin mRNA and protein. Unilateral infusion of hsiRNA-loaded exosomes, but not hsiRNAs alone, into mouse striatum resulted in bilateral oligonucleotide distribution and statistically significant bilateral silencing of up to 35% of Huntingtin mRNA. The broad distribution and efficacy of hsiRNA-loaded exosomes delivered to brain is expected to advance the development of therapies for the treatment of Huntington's disease and other neurodegenerative disorders.
递送是寡核苷酸疗法在治疗神经疾病（如亨廷顿舞蹈症）临床应用中的一个重大障碍。被称为外泌体的内源性小囊泡有潜力作为寡核苷酸递送载体，但目前缺乏将RNA治疗性物质载入外泌体的强大且可扩展的方法。在此，我们表明，经疏水修饰的小干扰RNA（hsiRNA）在共孵育时能有效载入外泌体，且不会改变囊泡的大小分布或完整性。装载了靶向亨廷顿蛋白mRNA的hsiRNA的外泌体被小鼠原代皮层神经元有效内化，并促进了亨廷顿蛋白mRNA和蛋白的剂量依赖性沉默。将装载hsiRNA的外泌体而非单独的hsiRNA单侧注入小鼠纹状体，导致寡核苷酸在双侧分布，且亨廷顿蛋白mRNA的双侧沉默率高达35%，具有统计学意义。预计递送至大脑的装载hsiRNA的外泌体的广泛分布和疗效将推动亨廷顿舞蹈症及其他神经退行性疾病治疗方法的发展。
