Nanoparticle-mediated gene transfer specific to retinal pigment epithelial cells
Nanoparticle-mediated gene transfer specific to retinal pigment epithelial cells
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DOI：
10.1016/j.biomaterials.2011.08.062
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发表时间：
2011-12-01
期刊：
影响因子：
14
通讯作者：
Naash, Muna I.
中科院分区：
文献类型：
--
作者：
Koirala, Adarsha;Makkia, Rasha S.;Naash, Muna I.
之前，我们在色素性视网膜炎模型中证明了ck3opeg10k压缩DNA纳米颗粒（NPs）有效地靶向光感受器细胞并改善视觉功能。在这里，我们使用RPE特异性报告载体（VMD2-eGFP）测试了这些NPs驱动视网膜色素上皮（RPE）中转基因表达的能力。NPs、未压实质粒或生理盐水经视网膜下递送至成年BALB/c小鼠。基于np的表达对RPE细胞具有特异性，对视网膜结构和功能无不良影响。在注射np后的第2天（PI-2）， eGFP表达水平达到峰值，稳定在接近于裸眼dna注射3倍的水平，并在最近的时间点（PI-30）保持升高。裸DNA只能转染注射部位的细胞，与裸DNA不同，NPs能够转染整个RPE细胞。视网膜下注射罗丹明标记的NPs和裸DNA显示相似的初始摄取到RPE细胞。然而，在PI-7和-30天，与裸DNA相比，注射np的眼睛RPE内检测到的荧光明显更多，这表明NPs在细胞内是稳定的，可能导致更高和持续的表达。总的来说，我们的研究结果表明，NPs可以有效地将基因传递到RPE，并且在治疗RPE相关疾病方面具有很大的潜力。Elsevier Ltd.出版。
Previously, we demonstrated that CK3OPEG10k-compacted DNA nanoparticles (NPs) efficiently target photoreceptor cells and improve visual function in a retinitis pigmentosa model. Here, we test the ability of these NPs in driving transgene expression in the retinal pigment epithelium (RPE), using an RPE-specific reporter vector (VMD2-eGFP). NPs, uncompacted plasmid, or saline were subretinally delivered to adult BALB/c mice. NP-based expression was specific to RPE cells and caused no deleterious effects on retinal structure and function. eGFP expression levels in NP-injected eyes peaked at post-injection day 2 (PI-2), stabilized at levels similar to 3-fold higher than in naked DNA-injected eyes, and remained elevated at the latest time-point examined (PI-30). Unlike naked DNA, which only transfected cells at the site of injection, NPs were able to transfect cells throughout the RPE. Subretinal injections of rhodamine labeled NPs and naked DNA showed comparable initial uptake into RPE cells. However, at PI-7 and -30 days significantly more fluorescence was detected inside the RPE of NP-injected eyes compared to naked DNA, suggesting NPs are stable inside the cell which could possibly lead to higher and sustained expression. Overall, our results demonstrate that NPs can efficiently deliver genes to the RPE and hold great potential for the treatment of RPE-associated diseases. Published by Elsevier Ltd.