Piperazine-derived lipid nanoparticles (LNP-A10) preferentially delivered mRNA to liver and splenic immune cells in vivo, achieving 50% functional delivery in Kupffer cells at a clinically relevant dose of 0.3 mg/kg.
Piperazine-derived lipid nanoparticles can preferentially deliver mRNA to hepatic and splenic immune cells in vivo without the need for active targeting ligands.
In humans, lipid nanoparticles (LNPs) have safely delivered therapeutic RNA to hepatocytes after systemic administration and to antigen-presenting cells after intramuscular injection. However, systemic RNA delivery to non-hepatocytes remains challenging, especially without targeting ligands such as antibodies, peptides, or aptamers. Here we report that piperazine-containing ionizable lipids (Pi-Lipids) preferentially deliver mRNA to immune cells in vivo without targeting ligands. After synthesizing and characterizing Pi-Lipids, we use high-throughput DNA barcoding to quantify how 65 chemically distinct LNPs functionally delivered mRNA (i.e., mRNA translated into functional, gene-editing protein) in 14 cell types directly in vivo. By analyzing the relationships between lipid structure and cellular targeting, we identify lipid traits that increase delivery in vivo. In addition, we characterize Pi-A10, an LNP that preferentially delivers mRNA to the liver and splenic immune cells at the clinically relevant dose of 0.3 mg/kg. These data demonstrate that high-throughput in vivo studies can identify nanoparticles with natural non-hepatocyte tropism and support the hypothesis that lipids with bioactive small-molecule motifs can deliver mRNA in vivo.
Ni et al. (Mon,) conducted a other in mRNA delivery to non-hepatocytes. Piperazine-derived lipid nanoparticles (LNP-A10) vs. PBS or unencapsulated DNA was evaluated on Percentage of tdTomato+ cells (functional mRNA delivery) in Kupffer cells at 0.3 mg/kg. Piperazine-derived lipid nanoparticles (LNP-A10) preferentially delivered mRNA to liver and splenic immune cells in vivo, achieving 50% functional delivery in Kupffer cells at a clinically relevant dose of 0.3 mg/kg.