Introduction

Lipid based nanoparticles (LNPs) are clinically successful vectors for hepatic delivery of nucleic acids. These systems are being developed for non-hepatic delivery of mRNA for the treatment of diseases like cystic fibrosis or retinal degeneration as well as infectious diseases. Localized delivery to the lungs requires aerosolization. We hypothesized that structural lipids within LNPs would provide features of integrity which can be tuned for attributes required for efficient hepatic and non-hepatic gene delivery. Herein, we explored whether naturally occurring lipids that originate from the cell membrane of plants and microorganisms enhance mRNA-based gene transfection in vitro and in vivo and whether they assist in maintaining mRNA activity after nebulization.

Methods

We substituted DSPC, a structural lipid used in a conventional LNP formulation, to a series of naturally occurring membrane lipids. We measured the effect of these membrane lipids on size, encapsulation efficiency and their impact on transfection efficiency. We further characterized LNPs after nebulization and measured whether they retained their transfection efficiency.

Results

One plant-derived structural lipid, DGTS, led to a significant improvement in liver transfection of mRNA. DGTS LNPs had similar transfection ability when administered in the nasal cavity to conventional LNPs. In contrast, we found that DGTS LNPs had reduced transfection efficiency in cells pre-and post-nebulization while maintaining size and encapsulation similar to DSPC LNPs.

Conclusions

We found that structural lipids provide differential mRNA-based activities in vitro and in vivo which also depend on the mode of administration. Understanding influence of structural lipids on nanoparticle morphology and structure can lead to engineering potent materials for mRNA-based gene therapy applications.

中文翻译：

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天然衍生的膜脂质影响基于纳米粒子的信使 RNA 传递。

介绍

基于脂质的纳米颗粒 (LNP) 是临床上成功的核酸肝递送载体。这些系统正在开发用于非肝脏递送 mRNA，以治疗囊性纤维化或视网膜变性等疾病以及传染病。局部输送到肺部需要雾化。我们假设 LNP 内的结构脂质将提供完整性特征，可以针对有效的肝脏和非肝脏基因传递所需的属性进行调整。在这里，我们探讨了源自植物和微生物细胞膜的天然脂质是否在体外和体内增强了基于 mRNA 的基因转染，以及它们是否有助于在雾化后维持 mRNA 活性。

方法

我们将传统 LNP 配方中使用的结构脂质 DSPC 替换为一系列天然存在的膜脂质。我们测量了这些膜脂对大小、封装效率及其对转染效率的影响。我们在雾化后进一步表征了 LNP，并测量了它们是否保留了转染效率。

结果

一种植物来源的结构脂质 DGTS 可显着改善 mRNA 的肝转染。DGTS LNPs 在鼻腔给药时与常规 LNPs 具有相似的转染能力。相比之下，我们发现 DGTS LNP 在雾化前后的细胞中降低了转染效率，同时保持了与 DSPC LNP 相似的大小和封装。

结论

我们发现结构脂质在体外和体内提供不同的基于 mRNA 的活性，这也取决于给药方式。了解结构脂质对纳米颗粒形态和结构的影响可以为基于 mRNA 的基因治疗应用设计有效的材料。