Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes.
Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes.
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DOI：
10.1016/j.jconrel.2015.08.007
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发表时间：
2015-11-10
期刊：
影响因子：
--
通讯作者：
Weissman D
中科院分区：
文献类型：
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作者：
Pardi N;Tuyishime S;Muramatsu H;Kariko K;Mui BL;Tam YK;Madden TD;Hope MJ;Weissman D
近年来，体外转录信使RNA(MRNA)已成为一种潜在的治疗平台。为了实现它的承诺，需要实现将mRNA有效地传递到特定类型的细胞和组织。脂质纳米粒(LNPs)是短干扰RNA的有效载体，目前已进入临床试验。然而，人们对LNPs传递信使核糖核酸的潜力知之甚少。在这里，我们通过将包含密码子优化的萤火虫荧光素酶的含有1-甲基伪尿苷的mRNA加入到稳定的LNPs中来产生mRNA-LNPs。小鼠通过6种不同途径注射0.005-0.250 mg/kg剂量的信使核糖核酸核糖核酸，通过体内成像可以检测到高水平的蛋白质翻译。皮下、肌肉和皮内注射注射部位局部翻译的LNP包裹的mRNA，最长可达10天。在接下来的几天里，通过气管内注射信使核糖核酸，可以在肺中获得高水平的蛋白质生产。静脉和腹膜内以及较小程度的肌肉和气管内递送导致了mRNA-LNPs的系统性运输，导致了mRNA在肝脏中的活跃翻译1-4天。我们的结果表明，LNPs是体内合适的信使核糖核酸载体，有可能成为运送编码治疗性蛋白的信使核糖核酸的有用工具。实验方法和数据获取的图形表示。数据的交付和分析方法在材料、方法和结果一节中介绍。
In recent years, in vitro transcribed messenger RNA (mRNA) has emerged as a potential therapeutic platform. To fulfill its promise, effective delivery of mRNA to specific cell types and tissues needs to be achieved. Lipid nanoparticles (LNPs) are efficient carriers for short-interfering RNAs and have entered clinical trials. However, little is known about the potential of LNPs to deliver mRNA. Here, we generated mRNA-LNPs by incorporating HPLC purified, 1-methylpseudouridine-containing mRNA comprising codon-optimized firefly luciferase into stable LNPs. Mice were injected with 0.005–0.250 mg/kg doses of mRNA-LNPs by 6 different routes and high levels of protein translation could be measured using in vivo imaging. Subcutaneous, intramuscular and intradermal injection of the LNP-encapsulated mRNA translated locally at the site of injection for up to 10 days. For several days, high levels of protein production could be achieved in the lung from the intratracheal administration of mRNA. Intravenous and intraperitoneal and to a lesser extent intramuscular and intratracheal deliveries led to trafficking of mRNA-LNPs systemically resulting in active translation of the mRNA in the liver for 1–4 days. Our results demonstrate that LNPs are appropriate carriers for mRNA in vivo and have the potential to become valuable tools for delivering mRNA encoding therapeutic proteins. Graphical representation of experimental approaches and data acquisition. Methods for the delivery and analysis of data are described in the Materials and Methods and Results sections.