Optimization of Lipid Nanoparticle Formulations for mRNA Delivery in Vivo with Fractional Factorial and Definitive Screening Designs
Optimization of Lipid Nanoparticle Formulations for mRNA Delivery in Vivo with Fractional Factorial and Definitive Screening Designs
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DOI：
10.1021/acs.nanolett.5b02497
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发表时间：
2015-11-01
期刊：
影响因子：
10.8
通讯作者：
Anderson, Daniel G.
中科院分区：
文献类型：
--
作者：
Kauffman, Kevin J.;Dorkin, J. Robert;Anderson, Daniel G.
信使RNA(信使RNA)的细胞内递送具有诱导蛋白质产生的潜力，用于许多治疗应用。尽管脂类纳米颗粒在传递小干扰RNA(SiRNA)方面显示出相当大的前景，但它们作为信使核糖核酸传递试剂的效用直到最近才被研究。最常见的siRNA制剂包含四种成分：含胺的脂质或类脂物质、磷脂、胆固醇和脂质锚定的聚乙二醇，它们的相对比例可能对制剂的效力产生深远的影响。在这里，我们开发了一种通用的策略，利用实验设计(DOE)方法，包括最终筛选和部分析因设计，优化脂质纳米粒配方，以便在体内将mRNA输送到肝脏。通过同时改变脂质比例和结构，我们开发了一种优化的配方，与以前用于siRNA递送的配方相比，该配方将负载促红细胞生成素mRNA的C12-200脂质纳米颗粒的效力提高了7倍。这一优化配方的关键特征是加入了1，2-二油酰基-3-甘油-3-磷代乙醇胺(DOPE)和增加了可电离脂质/信使核糖核酸重量比。有趣的是，优化的脂质纳米粒配方并没有改善siRNA的传递，这表明优化的配方参数设计对siRNA和mRNA的影响存在差异。我们相信，这里描述的通用方法可以加快具有大的多维设计空间的纳米颗粒配方的体内筛选和优化。
Intracellular delivery of messenger RNA (mRNA) has the potential to induce protein production for many therapeutic applications. Although lipid nanoparticles have shown considerable promise for the delivery of small interfering RNAs (siRNA), their utility as agents for mRNA delivery has only recently been investigated. The most common siRNA formulations contain four components: an amine-containing lipid or lipid-like material, phospholipid, cholesterol, and lipid-anchored polyethylene glycol, the relative ratios of which can have profound effects on the formulation potency. Here, we develop a generalized strategy to optimize lipid nanoparticle formulations for mRNA delivery to the liver in vivo using Design of Experiment (DOE) methodologies including Definitive Screening and Fractional Factorial Designs. By simultaneously varying lipid ratios and structures, we developed an optimized formulation which increased the potency of erythropoietin-mRNA-loaded C12-200 lipid nanoparticles 7-fold relative to formulations previously used for siRNA delivery. Key features of this optimized formulation were the incorporation of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and increased ionizable lipid:mRNA weight ratios. Interestingly, the optimized lipid nanoparticle formulation did not improve siRNA delivery, indicating differences in optimized formulation parameter design spates for siRNA and mRNA. We believe the general method described here can accelerate in vivo screening and optimization of nanoparticle formulations with large multidimensional design spaces.