Targeted Multifunctional Lipid ECO Plasmid DNA Nanoparticles as Efficient Non-viral Gene Therapy for Leber's Congenital Amaurosis.
Targeted Multifunctional Lipid ECO Plasmid DNA Nanoparticles as Efficient Non-viral Gene Therapy for Leber's Congenital Amaurosis.
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DOI：
10.1016/j.omtn.2017.02.005
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发表时间：
2017-06-16
期刊：
影响因子：
--
通讯作者：
Lu ZR
中科院分区：
文献类型：
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作者：
Sun D;Sahu B;Gao S;Schur RM;Vaidya AM;Maeda A;Palczewski K;Lu ZR
开发高效、安全的基因传递系统是基因治疗成功的关键。在这里，我们开发了一种使用多功能脂质ECO的靶向非病毒递送系统，用于治疗Leber先天性黑蒙2型（LCA 2），并在小鼠模型中进行了测试。ECO与质粒DNA（pDNA）在低胺/磷酸（N/P）比下形成稳定的纳米颗粒，并且在ARPE-19细胞中介导高的基因转染效率，这是因为ECO具有pH敏感的两亲性内体逃逸和还原性胞质释放（PERC）的内在特性。通过聚乙二醇（PEG）间隔物将与光感受器间类维生素A结合蛋白（IRBP）结合的全反式视黄胺掺入纳米颗粒中，以将pDNA靶向递送至视网膜色素上皮细胞中。靶向ECO/pDNA纳米颗粒在视网膜下注射后在1个月大的Rpe 65 −/−小鼠的RPE中提供高GFP表达。这些小鼠还表现出视网膜电图活性的显著增加，并且这种治疗效果持续至少120天。在野生型BALB/c小鼠中的安全性研究表明，视网膜下注射这些靶向纳米颗粒后没有不可逆的视网膜损伤。全反式-视黄胺修饰的ECO/pDNA纳米颗粒为安全有效地治疗RPE特异性单基因眼病如LCA 2提供了一个有前途的非病毒平台。
Development of a gene delivery system with high efficiency and a good safety profile is essential for successful gene therapy. Here we developed a targeted non-viral delivery system using a multifunctional lipid ECO for treating Leber’s congenital amaurosis type 2 (LCA2) and tested this in a mouse model. ECO formed stable nanoparticles with plasmid DNA (pDNA) at a low amine to phosphate (N/P) ratio and mediated high gene transfection efficiency in ARPE-19 cells because of their intrinsic properties of pH-sensitive amphiphilic endosomal escape and reductive cytosolic release (PERC). All-trans-retinylamine, which binds to interphotoreceptor retinoid-binding protein (IRBP), was incorporated into the nanoparticles via a polyethylene glycol (PEG) spacer for targeted delivery of pDNA into the retinal pigmented epithelium. The targeted ECO/pDNA nanoparticles provided high GFP expression in the RPE of 1-month-old Rpe65−/− mice after subretinal injection. Such mice also exhibited a significant increase in electroretinographic activity, and this therapeutic effect continued for at least 120 days. A safety study in wild-type BALB/c mice indicated no irreversible retinal damage following subretinal injection of these targeted nanoparticles. All-trans-retinylamine-modified ECO/pDNA nanoparticles provide a promising non-viral platform for safe and effective treatment of RPE-specific monogenic eye diseases such as LCA2.