Efficient nanoparticle mediated sustained RNA interference in human primary endothelial cells.
Efficient nanoparticle mediated sustained RNA interference in human primary endothelial cells.
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DOI：
10.1088/0957-4484/22/44/445101
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发表时间：
2011-11-04
期刊：
影响因子：
3.5
通讯作者：
Vishwanatha JK
中科院分区：
文献类型：
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作者：
Mukerjee A;Shankardas J;Ranjan AP;Vishwanatha JK
内皮细胞形成药物和/或基因治疗的重要靶标，因为内皮细胞在血管生成和血管功能中起关键作用，并且与各种病理生理学状况相关。RNA介导的基因沉默为克服许多此类疾病提供了一种新的治疗方法。但这种方法的主要挑战是确保shRNA对原代内皮细胞的最小毒性和有效转染效率。在本研究中，我们配制了shAnnexin A2负载PLGA纳米颗粒，其产生针对Annexin A2的细胞内siRNA，并导致Annexin A2的下调。发现纳米颗粒内质粒的包封百分比为57.65%。我们将我们的基于纳米颗粒的转染与脂质体介导的转染进行了比较，并且我们的研究表明，与原代视网膜微血管内皮细胞和人癌细胞系中的常规脂质体介导的转染相比，基于纳米颗粒的转染效率非常高（~97%）并且更持久。我们的研究结果还表明，装载有shAnnexin A2的PLGA纳米颗粒具有最小的毒性，在转染后24小时几乎95%的细胞是活的，而基于脂质体的转染仅产生30%的活细胞。因此，基于PLGA纳米颗粒的转染可用于对人原代内皮细胞和癌细胞的有效siRNA转染。这可能作为一个潜在的辅助治疗选择的疾病，如糖尿病视网膜病变，早产儿视网膜病变和年龄相关性黄斑变性除了各种癌症。
Endothelium forms an important target for drug and/or gene therapy since endothelial cells play critical roles in angiogenesis and vascular functions and are associated with various pathophysiological conditions. RNA-mediated gene silencing presents a new therapeutic approach to overcome many such diseases. But the major challenge of such an approach is to ensure minimal toxicity and effective transfection efficiency of shRNA to primary endothelial cells. In the present study, we formulated shAnnexin A2 loaded PLGA nanoparticles which produced intracellular siRNA against Annexin A2 and brought about the downregulation of Annexin A2. The percent encapsulation of the plasmid within the nanoparticle was found to be 57.65%. We compared our nanoparticle based transfections with lipofectamine mediated transfection and our studies show that nanoparticle based transfection efficiency is very high (~97%) and is more sustained as compared to conventional lipofectamine mediated transfections in primary retinal microvascular endothelial cells and human cancer cell lines. Our findings also show that the shAnnexin A2 loaded PLGA nanoparticles had minimal toxicity with almost 95% cells being viable 24 hours post transfection while lipofectamine based transfections resulted in only 30% viable cells. Therefore, PLGA nanoparticle based transfection may be used for efficient siRNA transfection to human primary endothelial and cancer cells. This may serve as a potential adjuvant treatment option for diseases such as diabetic retinopathy, retinopathy of prematurity and age related macular degeneration besides various cancers.
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影响因子：
--
作者：
Liu J;Rothermund CA;Ayala-Sanmartin J;Vishwanatha JK
通讯作者：
Vishwanatha JK
影响因子：
10.8
作者：
Gary, Dana J.;Puri, Nitin;Won, You-Yeon
通讯作者：
Won, You-Yeon
影响因子：
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作者：
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通讯作者：
Amoaku, WM
影响因子：
10.8
作者：
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通讯作者：
Ubrich, Nathalie
影响因子：
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作者：
Misra, Suniti;Hascall, Vincent C.;Ghatak, Shibnath
通讯作者：
Ghatak, Shibnath