Direct gene transfer with compacted DNA nanoparticles in retinal pigment epithelial cells: expression, repeat delivery and lack of toxicity
Direct gene transfer with compacted DNA nanoparticles in retinal pigment epithelial cells: expression, repeat delivery and lack of toxicity
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DOI：
10.2217/nnm.11.158
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发表时间：
2012-04-01
期刊：
影响因子：
5.5
通讯作者：
Naash, Muna I.
中科院分区：
文献类型：
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作者：
Han, Zongchao;Koirala, Adarsha;Naash, Muna I.
目的：评价致密DNA纳米颗粒（NPs）在视网膜色素上皮（RPE）细胞中的安全性。材料与方法：构建由rpe特异性黄斑营养不良启动子控制的增强GFP表达带和不带细菌骨架，并压实成用聚乙二醇取代赖氨酸30-mers配制的NPs。对成年BALB/c小鼠进行单次或双次视网膜下注射。通过视网膜电图检测注射后不同时间点GFP、促炎细胞因子、中性粒细胞/巨噬细胞介质的表达水平和视网膜功能。结果：免疫组织化学和实时荧光定量PCR显示NPs特异性转染RPE细胞的效率高于裸DNA，第二次注射后观察到类似的结果。注射后6小时，在所有队列中都观察到短暂的炎症反应，包括生理盐水，表明注射过程有不良反应。随后，所有实验组均未发现炎症反应。结论：本研究证明了np介导的RPE基因转移治疗在多次视网膜下给药后的安全性和有效性。
Aim: To evaluate the safety of compacted DNA nanoparticles (NPs) in retinal pigment epithelial (RPE) cells. Materials & methods: Enhanced GFP expression cassettes controlled by the RPE-specific vitelloform macular dystrophy promoter were constructed with and without a bacterial backbone and compacted into NPs formulated with polyethylene glycol-substituted lysine 30-mers. Single or double subretinal injections were administered in adult BALB/c mice. Expression levels of enhanced GFP, proinflammatory cytokines and neutrophil/macrophage mediators, and retinal function by electroretinogram were evaluated at different time-points postinjection. Results: Immunohistochemistry and real-time PCR demonstrated that NPs specifically transfect RPE cells at a higher efficiency than naked DNA and similar results were observed after the second injection. At 6 h postinjections, a transient inflammatory response was observed in all cohorts, including saline, indicating an adverse effect to the injection procedure. Subsequently, no inflammation was detected in all experimental groups. Conclusion: This study demonstrates the safety and efficacy of NP-mediated RPE gene transfer therapy following multiple subretinal administrations.