Lindsay Sarah, Hussain Muattaz, Binici Burcu, Perrie Yvonne
Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow G4 0RE, UK.
Vaccines (Basel). 2025 Mar 21;13(4):339. doi: 10.3390/vaccines13040339.
BACKGROUND/OBJECTIVES: The development of lipid nanoparticles (LNPs) as delivery platforms for nucleic acids has revolutionised possibilities for both therapeutic and vaccine applications. However, emerging studies highlight challenges in achieving reliable in vitro-in vivo correlation (IVIVC), which delays the translation of experimental findings into clinical applications. This study investigates these potential discrepancies by evaluating the physicochemical properties, in vitro efficacy (across three commonly used cell lines), and in vivo performance (mRNA expression and vaccine efficacy) of four LNP formulations.
LNPs composed of DSPC, cholesterol, a PEGylated lipid, and one of four ionizable lipids (SM-102, ALC-0315, MC3, or C12-200) were manufactured using microfluidics.
All formulations exhibited comparable physicochemical properties, as expected (size 70-100 nm, low PDI, near-neutral zeta potential, and high mRNA encapsulation). In vitro studies demonstrated variable LNP-mediated mRNA expression in both immortalised and immune cells, with SM-102 inducing significantly higher protein expression ( < 0.05) than the other formulations in immortalised and immune cells. However, in vivo results revealed that ALC-0315 and SM-102-based LNPs achieved significantly ( < 0.05) higher protein expression without a significant difference between them, while MC3- and C12-200-based LNPs exhibited lower expression levels. As vaccine formulations, all LNPs elicited strong immune responses with no significant differences among them.
These findings highlight the complexities of correlating in vitro and in vivo outcomes in LNP development and demonstrate the importance of holistic evaluation strategies to optimise their clinical translation.
背景/目的：脂质纳米颗粒（LNPs）作为核酸递送平台的发展，彻底改变了治疗和疫苗应用的可能性。然而，新出现的研究突出了实现可靠的体外-体内相关性（IVIVC）所面临的挑战，这延缓了实验结果向临床应用的转化。本研究通过评估四种LNP制剂的物理化学性质、体外疗效（在三种常用细胞系中）和体内性能（mRNA表达和疫苗效力）来研究这些潜在差异。
使用微流控技术制备由DSPC、胆固醇、聚乙二醇化脂质和四种可电离脂质之一（SM-102、ALC-0315、MC3或C12-200）组成的LNPs。
正如预期的那样，所有制剂都表现出可比的物理化学性质（尺寸70-100nm，低PDI，接近中性的zeta电位，以及高mRNA包封率）。体外研究表明，LNP介导的mRNA在永生化细胞和免疫细胞中的表达存在差异，在永生化细胞和免疫细胞中，SM-102诱导的蛋白表达显著高于其他制剂（<0.05）。然而，体内结果显示，基于ALC-0315和SM-102的LNPs实现了显著更高的蛋白表达（<0.05），且它们之间没有显著差异，而基于MC3和C12-200的LNPs表现出较低的表达水平。作为疫苗制剂，所有LNPs都引发了强烈的免疫反应，它们之间没有显著差异。
这些发现突出了LNP开发中体外和体内结果相关性的复杂性，并证明了整体评估策略对优化其临床转化的重要性。
