Gene silencing following siRNA delivery to skin via coated steel microneedles: In vitro and in vivo proof-of-concept.
Gene silencing following siRNA delivery to skin via coated steel microneedles: In vitro and in vivo proof-of-concept.
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DOI：
10.1016/j.jconrel.2012.12.030
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发表时间：
2013-03-28
期刊：
影响因子：
--
通讯作者：
Birchall JC
中科院分区：
文献类型：
--
作者：
Chong RH;Gonzalez-Gonzalez E;Lara MF;Speaker TJ;Contag CH;Kaspar RL;Coulman SA;Hargest R;Birchall JC
基于小干扰RNA（siRNA）的基因沉默疗法的发展，在有效治疗因基因表达异常导致的使人衰弱的遗传性、过度增殖性或恶性皮肤病方面具有巨大潜力。为了达到有效治疗效果并被医生和患者广泛接受，治疗性siRNA必须以稳定且具有功能的形式进入有活力的皮肤层，且最好给药过程无痛苦。在本研究中，我们探索了使用微创钢质微针装置将siRNA有效递送至皮肤。一种简单而精确的微针包被方法可实现将siRNA可重复地加载到单个微针上。从微针表面回收后，层粘连蛋白A/C（lamin A/C）的siRNA仍保持完全活性，这在HaCaT角质形成细胞中层粘连蛋白A/C的信使核糖核酸（mRNA）水平显著降低以及层粘连蛋白A/C蛋白减少中得到了证实。然而，与市售脂质转染试剂预复合的层粘连蛋白A/C的siRNA（siRNA脂质复合物）在微针包被后功能有所降低。由于针对CD44的Accell修饰的 “自我递送” siRNA在微针包被后也保留了功能，这种形式的siRNA被用于后续的体内研究，在转基因报告小鼠皮肤模型中测定基因沉默情况。将靶向报告基因（荧光素酶/绿色荧光蛋白）的自我递送siRNA包被在微针上并递送至小鼠足垫。对报告基因mRNA的定量分析以及对皮肤外层报告基因表达的活体成像证实，微针递送siRNA后在体内实现了功能性基因沉默。使用包被金属微针代表了一种全新、简单、微创、对患者友好且有可能自我给药的方法，可用于将治疗性核酸递送至皮肤。
The development of siRNA-based gene silencing therapies has significant potential for effectively treating debilitating genetic, hyper-proliferative or malignant skin conditions caused by aberrant gene expression. To be efficacious and widely accepted by physicians and patients, therapeutic siRNAs must access the viable skin layers in a stable and functional form, preferably without painful administration. In this study we explore the use of minimally-invasive steel microneedle devices to effectively deliver siRNA into skin. A simple, yet precise microneedle coating method permitted reproducible loading of siRNA onto individual microneedles. Following recovery from the microneedle surface, lamin A/C siRNA retained full activity, as demonstrated by significant reduction in lamin A/C mRNA levels and reduced lamin A/C protein in HaCaT keratinocyte cells. However, lamin A/C siRNA pre-complexed with a commercial lipid-based transfection reagent (siRNA lipoplex) was less functional following microneedle coating. As Accell-modified “self-delivery” siRNA targeted against CD44 also retained functionality after microneedle coating, this form of siRNA was used in subsequent in vivo studies, where gene silencing was determined in a transgenic reporter mouse skin model. Self-delivery siRNA targeting the reporter (luciferase/GFP) gene was coated onto microneedles and delivered to mouse footpad. Quantification of reporter mRNA and intravital imaging of reporter expression in the outer skin layers confirmed functional in vivo gene silencing following microneedle delivery of siRNA. The use of coated metal microneedles represents a new, simple, minimally-invasive, patient-friendly and potentially self-administrable method for the delivery of therapeutic nucleic acids to the skin.
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影响因子：
2.8
作者：
Haq, M. I.;Smith, E.;Birchall, J. C.
通讯作者：
Birchall, J. C.
影响因子：
5.7
作者：
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影响因子：
5.1
作者：
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影响因子：
3.6
作者：
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6.5
作者：
Hickerson, Robyn P.;Smith, Frances J. D.;Kaspar, Roger L.
通讯作者：
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