Piperazine-derived lipid nanoparticles deliver mRNA to immune cells in vivo

• Published in: Nature communications Vol. 13; no. 1; p. 4766
• Main Authors: Ni, Huanzhen, Hatit, Marine Z. C., Zhao, Kun, Loughrey, David, Lokugamage, Melissa P., Peck, Hannah E., Cid, Ada Del, Muralidharan, Abinaya, Kim, YongTae, Santangelo, Philip J., Dahlman, James E.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 15.08.2022; Nature Publishing Group; Nature Portfolio
• Subjects: 42; 42/109; 42/62; 49; 631/45/147; 631/61/2300; 631/61/350/354; 631/61/391; 639/925/926/1051; Antibodies; Antigen-presenting cells; Antigens; Aptamers; Cellular structure; Deoxyribonucleic acid; DNA; Gene sequencing; Genetic modification; Genome editing; Hepatocytes; Humanities and Social Sciences; Immune system; In vivo methods and tests; Ligands; Lipid structure; Lipids; mRNA; multidisciplinary; Nanoparticles; Peptides; Piperazine; Ribonucleic acid; RNA; Science; Science (multidisciplinary); Tropism
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-32281-5

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. Next-generation lipid nanoparticles that target non-hepatocytes could be important clinical tools. Using in vivo DNA barcoding, the authors identify piperazine-containing lipids deliver mRNA to immune cells without targeting ligands.