Expression kinetics of nucleoside-modified mRNA delivered in lipid nanoparticles to mice by various routes.
In recent years, in vitro transcribed messenger RNA (mRNA) has emerged as a potential therapeutic platform. To fulfill its promise, effective delivery of mRNA to specific cell types and tissues needs to be achieved. Lipid nanoparticles (LNPs) are efficient carriers for short-interfering RNAs and have entered clinical trials. However, little is known about the potential of LNPs to deliver mRNA. Here, we generated mRNA-LNPs by incorporating HPLC purified, 1-methylpseudouridine-containing mRNA comprising codon-optimized firefly luciferase into stable LNPs. Mice were injected with 0.005-0.250mg/kg doses of mRNA-LNPs by 6 different routes and high levels of protein translation could be measured using in vivo imaging. Subcutaneous, intramuscular and intradermal injection of the LNP-encapsulated mRNA translated locally at the site of injection for up to 10days. For several days, high levels of protein production could be achieved in the lung from the intratracheal administration of mRNA. Intravenous and intraperitoneal and to a lesser extent intramuscular and intratracheal deliveries led to trafficking of mRNA-LNPs systemically resulting in active translation of the mRNA in the liver for 1-4 days. Our results demonstrate that LNPs are appropriate carriers for mRNA in vivo and have the potential to become valuable tools for delivering mRNA encoding therapeutic proteins.
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Norbert
N
Pardi
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Steven
S
Tuyishime
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Hiromi
H
Muramatsu
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Katalin
K
Kariko
Acuitas Therapeutics, Vancouver, V6T 1Z3 BC, Canada.
Barbara L
BL
Mui
Acuitas Therapeutics, Vancouver, V6T 1Z3 BC, Canada.
Ying K
YK
Tam
Acuitas Therapeutics, Vancouver, V6T 1Z3 BC, Canada.
Thomas D
TD
Madden
Acuitas Therapeutics, Vancouver, V6T 1Z3 BC, Canada.
Michael J
MJ
Hope
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: dreww@mail.med.upenn.edu.
Drew
D
Weissman
D000818:Animals; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000378:metabolism; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism*; D008168:Lung / Q000378:metabolism; D008807:Mice, Inbred BALB C; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D011560:Pseudouridine / Q000031:analogs & derivatives* / Q000737:chemistry; D012333:RNA, Messenger / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics; D014162:Transfection / Q000379:methods
D010714:Phosphatidylethanolamines; D012333:RNA, Messenger; C013608:1-methylpseudouridine; D011560:Pseudouridine; C020888:1,2-dielaidoylphosphatidylethanolamine; D049409:Luciferases, Firefly
Luciferase; Nanoparticle; Non-viral gene delivery; Pseudouridine; mRNA