Combinatorial development of nebulized mRNA delivery formulations for the lungs

Nat Nanotechnol. 2024 Mar;19(3):364-375. doi: 10.1038/s41565-023-01548-3. Epub 2023 Nov 20.


Inhaled delivery of mRNA has the potential to treat a wide variety of diseases. However, nebulized mRNA lipid nanoparticles (LNPs) face several unique challenges including stability during nebulization and penetration through both cellular and extracellular barriers. Here we develop a combinatorial approach addressing these barriers. First, we observe that LNP formulations can be stabilized to resist nebulization-induced aggregation by altering the nebulization buffer to increase the LNP charge during nebulization, and by the addition of a branched polymeric excipient. Next, we synthesize a combinatorial library of ionizable, degradable lipids using reductive amination, and evaluate their delivery potential using fully differentiated air-liquid interface cultured primary lung epithelial cells. The final combination of ionizable lipid, charge-stabilized formulation and stability-enhancing excipient yields a significant improvement in lung mRNA delivery over current state-of-the-art LNPs and polymeric nanoparticles.

MeSH terms

  • Cell Differentiation
  • Excipients*
  • Nanoparticles*
  • Polymers
  • RNA, Messenger / genetics
  • RNA, Small Interfering


  • Excipients
  • Polymers
  • RNA, Messenger
  • RNA, Small Interfering