PDF(Pubmed)
Abstract：
Lipid nanoparticles (LNPs) tailored for mRNA delivery were optimized to serve as a platform for treating metabolic diseases. Four distinct lipid mixes (LMs) were formulated by modifying various components: LM1 (ALC-0315/DSPC/Cholesterol/ALC-0159), LM2 (ALC-0315/DOPE/Cholesterol/ALC-0159), LM3 (ALC-0315/DSPC/Cholesterol/DMG-PEG2k), and LM4 (DLin-MC3-DMA/DSPC/Cholesterol/ALC-0159). LNPs exhibited stability and homogeneity with a mean size of 75 to 90 nm, confirmed by cryo-TEM and SAXS studies. High mRNA encapsulation (95-100%) was achieved. LNPs effectively delivered EGFP-encoding mRNA to HepG2 and DC2.4 cell lines. LNPs induced cytokine secretion from human peripheral blood mononuclear cells (PBMCs), revealing that LM1, LM2, and LM4 induced 1.5- to 4-fold increases in IL-8, TNF-α, and MCP-1 levels, while LM3 showed minimal changes. Reporter mRNA expression was observed in LNP-treated PBMCs. Hemotoxicity studies confirmed formulation biocompatibility with values below 2%. In vivo biodistribution in mice post intramuscular injection showed significant mRNA expression, mainly in the liver. The modification of LNP components influenced reactogenicity, inflammatory response, and mRNA expression, offering a promising platform for selecting less reactogenic carriers suitable for repetitive dosing in metabolic disease treatment.
摘要：
针对mRNA递送定制的脂质纳米颗粒(LNP)被优化以用作治疗代谢疾病的平台。四种不同的脂质混合物（LM）通过修饰各种成分配制：LM1（ALC-0315/DSPC/胆固醇/ALC-0159），LM2（ALC-0315/DOPE/胆固醇/ALC-0159），LM3（ALC-0315/DSPC/胆固醇/DMG-PEG2k），和LM4(DLin-MC3-DMA/DSPC/胆固醇/ALC-0159)。LNP表现出稳定性和均匀性，平均尺寸为75至90nm，通过低温TEM和SAXS研究证实。实现了高mRNA包封(95-100%)。LNP有效地将编码EGFP的mRNA递送至HepG2和DC2.4细胞系。LNPs诱导人外周血单核细胞（PBMC）分泌细胞因子，揭示LM1、LM2和LM4诱导IL-8、TNF-α增加1.5到4倍，和MCP-1级别，而LM3显示最小变化。在LNP处理的PBMC中观察到报告mRNA表达。血液毒性研究证实制剂生物相容性的值低于2%。肌肉注射后小鼠体内生物分布显示显著的mRNA表达，主要在肝脏。LNP组分的修饰影响了反应原性，炎症反应，和mRNA表达，为选择适用于代谢疾病治疗中重复给药的反应基因较少的载体提供了一个有前途的平台。
