Linkage between endosomal escape of LNP-mRNA and loading into EVs for transport to other cells

Nat Commun. 2019 Sep 24;10(1):4333. doi: 10.1038/s41467-019-12275-6.

Abstract

RNA-based therapeutics hold great promise for treating diseases and lipid nanoparticles (LNPs) represent the most advanced platform for RNA delivery. However, the fate of the LNP-mRNA after endosome-engulfing and escape from the autophagy-lysosomal pathway remains unclear. To investigate this, mRNA (encoding human erythropoietin) was delivered to cells using LNPs, which shows, for the first time, a link between LNP-mRNA endocytosis and its packaging into extracellular vesicles (endo-EVs: secreted after the endocytosis of LNP-mRNA). Endosomal escape of LNP-mRNA is dependent on the molar ratio between ionizable lipids and mRNA nucleotides. Our results show that fractions of ionizable lipids and mRNA (1:1 molar ratio of hEPO mRNA nucleotides:ionizable lipids) of endocytosed LNPs were detected in endo-EVs. Importantly, these EVs can protect the exogenous mRNA during in vivo delivery to produce human protein in mice, detected in plasma and organs. Compared to LNPs, endo-EVs cause lower expression of inflammatory cytokines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Transport
  • Cell Line
  • Cytoplasm / metabolism
  • Endosomes / metabolism
  • Endosomes / physiology*
  • Endosomes / ultrastructure
  • Erythropoietin / genetics
  • Erythropoietin / metabolism*
  • Extracellular Vesicles / metabolism*
  • Female
  • Humans
  • Lipid Metabolism*
  • Lipids / chemistry
  • Mice
  • Mice, Inbred C57BL
  • Nanoparticles / chemistry
  • Nanoparticles / metabolism*
  • RNA, Messenger / metabolism*

Substances

  • EPO protein, human
  • Lipids
  • RNA, Messenger
  • Erythropoietin