Pharmacodelivery Group, School of Pharmacy, University College Cork , Cork, Ireland.
Mol Pharm. 2013 Feb 4;10(2):640-9. doi: 10.1021/mp3003946. Epub 2013 Jan 9.
Huntington's disease (HD) is a rare autosomal dominant neurodegenerative disease caused by the expression of a toxic Huntingtin (HTT) protein. The use of short interfering RNAs (siRNAs) to silence the mutant protein is one of the most promising therapeutic strategies under investigation. The biggest caveat to siRNA-based approaches is the lack of efficient and nontoxic delivery vectors for siRNA delivery to the central nervous system. In this study, we investigated the potential of modified amphiphilic β-cyclodextrins (CDs), oligosaccharide-based molecules, as novel siRNA neuronal carriers. We show that CDs formed nanosize particles which were stable in artificial cerebrospinal fluid. Moreover, these complexes were able to reduce the expression of the HTT gene in rat striatal cells (ST14A-HTT120Q) and in human HD primary fibroblasts. Only limited toxicity was observed with CD·siRNA nanoparticles in any of the in vitro models used. Sustained knockdown effects were observed in the striatum of the R6/2 mouse model of HD after single direct injections of CD·siRNA nanoparticles. Repeated brain injections of CD·siRNA complexes resulted in selective alleviation of motor deficits in this mouse model. Together these data support the utility of modified β-CDs as efficient and safe siRNA delivery vectors for RNAi-based therapies for neuropsychiatric and neurodegenerative disorders.
亨廷顿病(HD)是一种罕见的常染色体显性神经退行性疾病,由毒性亨廷顿蛋白(HTT)的表达引起。使用短干扰 RNA(siRNA)沉默突变蛋白是正在研究的最有前途的治疗策略之一。基于 siRNA 的方法最大的缺点是缺乏有效的、非毒性的用于 siRNA 递送至中枢神经系统的载体。在这项研究中,我们研究了经过修饰的两亲性β-环糊精(CDs)的潜力,这是一种基于寡糖的分子,作为新型 siRNA 神经元载体。我们表明,CD 形成纳米尺寸的颗粒,在人工脑脊液中稳定。此外,这些复合物能够减少大鼠纹状体细胞(ST14A-HTT120Q)和人 HD 原代成纤维细胞中 HTT 基因的表达。在任何使用的体外模型中,CD·siRNA 纳米颗粒都只观察到有限的毒性。在 R6/2 型 HD 小鼠模型中,单次直接注射 CD·siRNA 纳米颗粒后,纹状体中可观察到持续的基因敲低效应。CD·siRNA 复合物的重复脑内注射导致该小鼠模型的运动缺陷选择性缓解。这些数据共同支持了经过修饰的β-CDs 作为用于神经精神和神经退行性疾病的 RNAi 治疗的有效和安全的 siRNA 递送载体的实用性。