Xue Lulu, Xiong Xinhong, Zhao Gan, Molina-Arocho William, Palanki Rohan, Xiao Zebin, Han Xuexiang, Yoon Il-Chul, Figueroa-Espada Christian G, Xu Junchao, Gong Ningqiang, Shi Qiangqiang, Chen Qinyuan, Alameh Mohamad-Gabriel, Vaughan Andrew E, Haldar Malay, Wang Karin, Weissman Drew, Mitchell Michael J
Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
Yangtze Delta Region Institute (Huzhou), University of Electronic Science and Technology of China, Huzhou, Zhejiang 313001, China.
J Am Chem Soc. 2025 Jan 15;147(2):1542-1552. doi: 10.1021/jacs.4c10265. Epub 2025 Jan 1.
Lipid nanoparticles (LNPs) have emerged as pivotal vehicles for messenger RNA (mRNA) delivery to hepatocytes upon systemic administration and to antigen-presenting cells following intramuscular injection. However, achieving systemic mRNA delivery to non-hepatocytes remains challenging without the incorporation of targeting ligands such as antibodies, peptides, or small molecules. Inspired by comb-like polymeric architecture, here we utilized a multiarm-assisted design to construct a library of 270 dendron-like degradable ionizable lipids by altering the structures of amine heads and multiarmed tails for optimal mRNA delivery. Following in vitro high-throughput screening, a series of top-dendron-like LNPs with high transfection efficacy were identified. These dendron-like ionizable lipids facilitated greater mRNA delivery to the spleen in vivo compared to ionizable lipid analogs lacking dendron-like structure. Proteomic analysis of corona-LNP pellets showed enhancement of key protein clusters, suggesting potential endogenous targeting to the spleen. A lead dendron-like LNP formulation, 18-2-9b2, was further used to encapsulate Cre mRNA and demonstrated excellent genome modification in splenic macrophages, outperforming a spleen-tropic MC3/18PA LNP in the Ai14 mice model. Moreover, 18-2-9b2 LNP encapsulating therapeutic BTB domain and CNC homologue 1 (BACH1) mRNA exhibited proficient BACH1 expression and subsequent Spic downregulation in splenic red pulp macrophages (RPM) in a Spic-GFP transgene model upon intravenous administration. These results underscore the potential of dendron-like LNPs to facilitate mRNA delivery to splenic macrophages, potentially opening avenues for a range of mRNA-LNP therapeutic applications, including regenerative medicine, protein replacement, and gene editing therapies.
脂质纳米颗粒（LNPs）已成为信使核糖核酸（mRNA）全身给药时递送至肝细胞以及肌肉注射后递送至抗原呈递细胞的关键载体。然而，在不掺入靶向配体（如抗体、肽或小分子）的情况下，实现mRNA全身递送至非肝细胞仍然具有挑战性。受梳状聚合物结构的启发，我们利用多臂辅助设计，通过改变胺头和多臂尾部的结构，构建了一个包含270种树枝状可降解可电离脂质的文库，以实现最佳的mRNA递送。经过体外高通量筛选，鉴定出了一系列具有高转染效率的顶级树枝状LNPs。与缺乏树枝状结构的可电离脂质类似物相比，这些树枝状可电离脂质在体内促进了更多的mRNA递送至脾脏。对冠-LNP沉淀的蛋白质组学分析显示关键蛋白簇有所增强，表明可能存在对脾脏的内源性靶向作用。一种先导树枝状LNP制剂18-2-9b2被进一步用于封装Cre mRNA，并在Ai14小鼠模型中证明在脾巨噬细胞中具有出色的基因组修饰能力，优于脾脏靶向的MC3/18PA LNP。此外，在Spic-GFP转基因模型中，静脉注射时，封装治疗性BTB结构域和CNC同源物1（BACH1）mRNA的18-2-9b2 LNP在脾红髓巨噬细胞（RPM）中表现出高效的BACH1表达以及随后的Spic下调。这些结果强调了树枝状LNPs促进mRNA递送至脾巨噬细胞中的潜力，可能为一系列mRNA-LNP治疗应用开辟道路，包括再生医学、蛋白质替代和基因编辑疗法。
