Cheng Miffy Hok Yan, Zhang Yao, Fox Kevin, Leung Jerry, Strong Colton, Kang Emma, Chen Yihang, Tong Michelle, Bommadevara Hemashree, Jan Eric, Ip Owen Yuk Long, Rodríguez-Rodríguez Cristina, Saatchi Katayoun, Häfeli Urs O, Abdolahzadeh Amir, Witzigmann Dominik, Cullis Pieter R
Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, Canada.
Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada.
Nat Commun. 2025 May 3;16(1):4135. doi: 10.1038/s41467-025-58523-w.
Long-circulating, transfection-competent lipid nanoparticle (LNP)-mRNA delivery systems are critical for achieving efficient transfection in extrahepatic tissues. Here we investigate the properties of LNP mRNA systems containing high proportions of bilayer forming lipids, using equimolar egg sphingomyelin and cholesterol as the bilayer-forming components. We show that LNP mRNA systems prepared at bilayer lipid to ionizable lipid molar ratios of 4-0.67 exhibit high mRNA encapsulation efficiencies (90-100%) and excellent transfection potencies in vitro. Systems with bilayer lipid to ionizable lipid molar ratios equating to 4 exhibit a liposomal morphology with a solid core suspended in an aqueous interior surrounded by a lipid bilayer. These liposomal LNPs exhibit longer circulation lifetimes than LNP systems with Onpattro-like lipid compositions and have enhanced extrahepatic transfection properties. The prolonged blood circulation lifetime is attributed to reduced plasma protein adsorption. The transfection competency of liposomal LNP systems is attributed to export of the solid core containing mRNA from the LNP as the endosomal pH is lowered.
长效循环、具备转染能力的脂质纳米颗粒（LNP）-mRNA递送系统对于在肝外组织中实现高效转染至关重要。在此，我们使用等摩尔的蛋黄鞘磷脂和胆固醇作为形成双层的成分，研究了含有高比例形成双层脂质的LNP mRNA系统的性质。我们发现，以双层脂质与可电离脂质的摩尔比为4-0.67制备的LNP mRNA系统在体外表现出高mRNA包封效率（90-100%）和出色的转染能力。双层脂质与可电离脂质摩尔比等于4的系统呈现出脂质体形态，其固体核心悬浮在由脂质双层包围的水性内部。这些脂质体LNP比具有Onpattro样脂质组成的LNP系统具有更长的循环寿命，并具有增强的肝外转染特性。血液循环寿命的延长归因于血浆蛋白吸附的减少。脂质体LNP系统的转染能力归因于随着内体pH值降低，含有mRNA的固体核心从LNP中输出。
