Journal Article

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

Hölttä, Mikko, Johansson, Svante, Lindfors, Lennart, Mårtensson, Inga-Lill, Jin, Tao, Valadi, Hadi, Camponeschi, Alessandro, Sundqvist, Martina, Kjellman, Tomas, Nawaz, Muhammad, Papadimitriou, Alexandros, Angerfors, Annelie, Lindquist, Johnny, Na, Manli, Maugeri, Marco, Östman, Sofia, Skantze, Pia, Sunnerhagen, Per

2019

Overview

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. Lipid nanoparticles (LNPs) are potential platforms for RNA-based therapeutics, but the fate of LNP-RNAs upon internalization into the cell is unclear. Here, the authors show that LNP-mRNAs and ionizable lipids escape the endosomes and are re-released via extracellular vesicles which could deliver the functional mRNA to other cells.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 13/1 / 13/106 / 13/109 / 13/21 / 13/31 / 38 / 38/1 / 38/109 / 38/77 / 38/88 / 38/90 / 42/109 / 42/34 / 631/61/2300 / 631/80/642/1776 / 64/60 / 82/1 / 82/16 / 82/80 / 82/83 / Animals / Autophagy / Basic Medicine / Biological Transport / Cell Line / Cytokines / Cytoplasm - metabolism / Endocytosis / Endosomes - metabolism / Endosomes - physiology / Endosomes - ultrastructure / Erythropoietin / Erythropoietin - genetics / Erythropoietin - metabolism / Exosomes / Extracellular vesicles / Extracellular Vesicles - metabolism / Female / Humanities and Social Sciences / Humans / Inflammation / Lipid Metabolism / Lipids / Lipids - chemistry / Medical Biotechnology (Focus on Cell Biology (incl. Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) / Medical treatment / Medicinsk bioteknologi (Inriktn. mot cellbiologi (inkl. stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci) / Medicinska och farmaceutiska grundvetenskaper / Mice / Mice, Inbred C57BL / mRNA / multidisciplinary / Nanoparticles / Nanoparticles - chemistry / Nanoparticles - metabolism / Nucleotides / Organs / Phagocytosis / RNA transport / RNA, Messenger - metabolism / Science / Science (multidisciplinary)