Boosting intracellular delivery of lipid nanoparticle-encapsulated messenger RNA

Intracellular delivery of mRNA holds great potential for vaccine 1 – 3 and therapeutic 4 discovery and development. Despite increasing recognition of the utility of lipid-based nanoparticles (LNPs) for intracellular delivery of mRNA, particle engineering is hindered by insufficient understanding of...

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Published in:Nano letters 2017-08, Vol.17 (9), p.5711-5718
Main Authors: Patel, Siddharth, Ashwanikumar, N, Robinson, Emily, DuRoss, Allison, Sun, Conroy, Murphy-Benenato, Kerry E., Mihai, Cosmin, Almarsson, Örn, Sahay, Gaurav
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container_issue 9
container_start_page 5711
container_title Nano letters
container_volume 17
creator Patel, Siddharth
Ashwanikumar, N
Robinson, Emily
DuRoss, Allison
Sun, Conroy
Murphy-Benenato, Kerry E.
Mihai, Cosmin
Almarsson, Örn
Sahay, Gaurav
description Intracellular delivery of mRNA holds great potential for vaccine 1 – 3 and therapeutic 4 discovery and development. Despite increasing recognition of the utility of lipid-based nanoparticles (LNPs) for intracellular delivery of mRNA, particle engineering is hindered by insufficient understanding of endosomal escape, which is believed to be a main limiter of cytosolic availability and activity of the nucleic acid inside the cell. Using a series of CRISPR-based genetic perturbations of the lysosomal pathway, we have identified that late endosome/lysosome (LE/Ly) formation is essential for functional delivery of exogenously presented mRNA. Lysosomes provide a spatio-temporal hub to orchestrate mTOR signaling and are known to control cell proliferation, nutrient sensing, ribosomal biogenesis, and mRNA translation. Through modulation of the mTOR pathway we were able to enhance or inhibit LNP-mediated mRNA delivery. To further boost intracellular delivery of mRNA we screened 212 bioactive lipid-like molecules that are either enriched in vesicular compartments or modulate cell signaling. Surprisingly, we have discovered that leukotriene-antagonists, clinically approved for treatment of asthma and other lung diseases, enhance intracellular mRNA delivery in vitro (over 3-fold, p
doi_str_mv 10.1021/acs.nanolett.7b02664
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Surprisingly, we have discovered that leukotriene-antagonists, clinically approved for treatment of asthma and other lung diseases, enhance intracellular mRNA delivery in vitro (over 3-fold, p&lt;0.005) and in vivo (over 2-fold, p&lt;0.005). 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title Boosting intracellular delivery of lipid nanoparticle-encapsulated messenger RNA
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