Sakers Sophia H, Fiduccia Gianna, Byrne Katherine E, Reddy B Pradeep K, Dahlman James E, Prausnitz Mark R
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA, USA.
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA.
Eur J Pharm Biopharm. 2025 Aug 8:114819. doi: 10.1016/j.ejpb.2025.114819.
Messenger RNA (mRNA) encapsulated in lipid nanoparticles (LNPs) is a potent technology with broad applications. Microneedle patches (MNPs) can enhance the accessibility of mRNA-LNPs for vaccination or therapeutic applications. We evaluated the effects of LNP composition on the stability of mRNA-LNPs before and after MNP manufacturing, as assessed by changes in mRNA-LNP size, encapsulation efficiency, and protein expression in vitro and in vivo. The lipid molar ratio had a significant impact on LNP characteristics and ability to withstand stressors of MNP fabrication. An elevation in ionizable lipid content from 50% to 60% increased in vitro mRNA expression, measured by luciferase expression in RAW 264.7 cells, following extraction from an MNP. Among the three ionizable lipids tested, SM-102 had the highest expression before and after MNP manufacturing. Cholesterol analogues influenced in vivo expression of mRNA-LNPs before MNP manufacturing, but the effect was absent in mRNA-LNP MNPs. PEGylated lipid choice affected mRNA encapsulation in LNPs and had a strong effect on in vitro expression, but this effect was not seen in vivo. Phospholipid choice did not have a significant impact on most mRNA-LNP characteristics, but the use of DOTMA increased in vitro expression. These data suggest that LNP composition can affect mRNA-LNP characteristics and function both before and after MNP manufacturing. Future mRNA-LNP MNP designs should consider the effect of lipid molar ratios and favor higher ionizable lipid content.
包裹在脂质纳米颗粒（LNP）中的信使核糖核酸（mRNA）是一项具有广泛应用前景的强大技术。微针贴片（MNP）可提高mRNA-LNP在疫苗接种或治疗应用中的可及性。我们评估了LNP组成对MNP制造前后mRNA-LNP稳定性的影响，通过mRNA-LNP大小、包封效率以及体外和体内蛋白质表达的变化进行评估。脂质摩尔比对LNP特性以及承受MNP制造压力的能力有显著影响。从MNP中提取后，在RAW 264.7细胞中通过荧光素酶表达测量，可电离脂质含量从50%提高到60%会增加体外mRNA表达。在所测试的三种可电离脂质中，SM-102在MNP制造前后的表达最高。胆固醇类似物在MNP制造前影响mRNA-LNP的体内表达，但在mRNA-LNP MNP中没有这种影响。聚乙二醇化脂质的选择影响mRNA在LNP中的包封，并对体外表达有强烈影响，但在体内未观察到这种影响。磷脂的选择对大多数mRNA-LNP特性没有显著影响，但使用DOTMA会增加体外表达。这些数据表明，LNP组成在MNP制造前后均可影响mRNA-LNP的特性和功能。未来mRNA-LNP MNP的设计应考虑脂质摩尔比的影响，并倾向于更高的可电离脂质含量。
