Anionic Lipid Nanoparticles Preferentially Deliver mRNA to the Hepatic Reticuloendothelial System

Roy Pattipeiluhu; Gabriela Arias-Alpizar; Genc Basha; Karen Y.T. Chan; Jeroen Bussmann; Thomas H. Sharp; Mohammad Amin Moradi; Nico Sommerdijk; Edward N. Harris; Pieter R. Cullis; Alexander Kros; Dominik Witzigmann; Frederick Campbell

doi:10.1002/adma.202201095

Anionic Lipid Nanoparticles Preferentially Deliver mRNA to the Hepatic Reticuloendothelial System

Roy Pattipeiluhu
, Gabriela Arias-Alpizar
, Genc Basha
, Karen Y.T. Chan
, Jeroen Bussmann
, Thomas H. Sharp
, Mohammad Amin Moradi
, Nico Sommerdijk
, Edward N. Harris
, Pieter R. Cullis
, Alexander Kros
, Dominik Witzigmann (Corresponding author)
, Frederick Campbell (Corresponding author)

Research output: Contribution to journal › Article › Academic › peer-review

189 Citations (Scopus)

541 Downloads (Pure)

Abstract

Lipid nanoparticles (LNPs) are the leading nonviral technologies for the delivery of exogenous RNA to target cells in vivo. As systemic delivery platforms, these technologies are exemplified by Onpattro, an approved LNP-based RNA interference therapy, administered intravenously and targeted to parenchymal liver cells. The discovery of systemically administered LNP technologies capable of preferential RNA delivery beyond hepatocytes has, however, proven more challenging. Here, preceded by comprehensive mechanistic understanding of in vivo nanoparticle biodistribution and bodily clearance, an LNP-based messenger RNA (mRNA) delivery platform is rationally designed to preferentially target the hepatic reticuloendothelial system (RES). Evaluated in embryonic zebrafish, validated in mice, and directly compared to LNP–mRNA systems based on the lipid composition of Onpattro, RES-targeted LNPs significantly enhance mRNA expression both globally within the liver and specifically within hepatic RES cell types. Hepatic RES targeting requires just a single lipid change within the formulation of Onpattro to switch LNP surface charge from neutral to anionic. This technology not only provides new opportunities to treat liver-specific and systemic diseases in which RES cell types play a key role but, more importantly, exemplifies that rational design of advanced RNA therapies must be preceded by a robust understanding of the dominant nano–biointeractions involved.

Original language	English
Article number	2201095
Number of pages	15
Journal	Advanced Materials
Volume	34
Issue number	16
DOIs	https://doi.org/10.1002/adma.202201095
Publication status	Published - 21 Apr 2022

Bibliographical note

Funding Information:
R.P. and G.A.‐A. contributed equally to this work. Panagiota Papadopoulou (Leiden University) is thanked for her work in developing apoE‐targeted liposomes within the embryonic zebrafish. Niek Crone (Leiden University) is thanked for gifting 3‐azido‐5‐cholestene. This work was supported financially by the Netherlands Organization for Scientific Research (NWO‐VICI – Project No. 724.014.001) – R.P., G.A.‐A.; a M‐ERA Grant – F.C., M.‐A.M.; the Interreg 2 Seas Program 2014–2020 co‐funded by the European Regional Development Fund under subsidy contract “Site Drug 2S07‐033” – G.A.‐A.; the NanoMedicines Innovation Network (NMIN) – G.B., D.W.; and the Swiss National Science Foundation (Grant No. 186741) – D.W. Breeding and housing of KO mice was supported by the National Institute of Health (Grant No. HL130864) – E.N.H. All mouse protocols were approved by the Canadian Animal Care Committee and conducted in accordance with relevant guidelines and regulations. Zebrafish (Danio rerio, strain AB/TL) were maintained and handled according to the guidelines from the Zebrafish Model Organism Database (http://zfin.org) and in compliance with the directives of the local animal welfare committee of Leiden University. stab2 −/−

Funding

R.P. and G.A.-A. contributed equally to this work. Panagiota Papadopoulou (Leiden University) is thanked for her work in developing apoE-targeted liposomes within the embryonic zebrafish. Niek Crone (Leiden University) is thanked for gifting 3-azido-5-cholestene. This work was supported financially by the Netherlands Organization for Scientific Research (NWO-VICI ? Project No. 724.014.001) ? R.P., G.A.-A.; a M-ERA Grant ? F.C., M.-A.M.; the Interreg 2 Seas Program 2014?2020 co-funded by the European Regional Development Fund under subsidy contract ?Site Drug 2S07-033? ? G.A.-A.; the NanoMedicines Innovation Network (NMIN) ? G.B., D.W.; and the Swiss National Science Foundation (Grant No. 186741) ? D.W. Breeding and housing of stab2?/? KO mice was supported by the National Institute of Health (Grant No. HL130864) ? E.N.H. All mouse protocols were approved by the Canadian Animal Care Committee and conducted in accordance with relevant guidelines and regulations. Zebrafish (Danio rerio, strain AB/TL) were maintained and handled according to the guidelines from the Zebrafish Model Organism Database (http://zfin.org) and in compliance with the directives of the local animal welfare committee of Leiden University. R.P. and G.A.‐A. contributed equally to this work. Panagiota Papadopoulou (Leiden University) is thanked for her work in developing apoE‐targeted liposomes within the embryonic zebrafish. Niek Crone (Leiden University) is thanked for gifting 3‐azido‐5‐cholestene. This work was supported financially by the Netherlands Organization for Scientific Research (NWO‐VICI – Project No. 724.014.001) – R.P., G.A.‐A.; a M‐ERA Grant – F.C., M.‐A.M.; the Interreg 2 Seas Program 2014–2020 co‐funded by the European Regional Development Fund under subsidy contract “Site Drug 2S07‐033” – G.A.‐A.; the NanoMedicines Innovation Network (NMIN) – G.B., D.W.; and the Swiss National Science Foundation (Grant No. 186741) – D.W. Breeding and housing of KO mice was supported by the National Institute of Health (Grant No. HL130864) – E.N.H. All mouse protocols were approved by the Canadian Animal Care Committee and conducted in accordance with relevant guidelines and regulations. Zebrafish (Danio rerio, strain AB/TL) were maintained and handled according to the guidelines from the Zebrafish Model Organism Database (http://zfin.org) and in compliance with the directives of the local animal welfare committee of Leiden University. stab2 −/−

Keywords

embryonic zebrafish
lipid nanoparticles
mRNA delivery
reticuloendothelial system
stabilin-2
Mononuclear Phagocyte System/metabolism
RNA, Small Interfering/genetics
Lipids
Zebrafish
RNA, Messenger/metabolism
Nanoparticles
Tissue Distribution
Animals
Liver/metabolism
Mice
Liposomes

Access to Document

10.1002/adma.202201095

pdfFinal published version, 5.03 MBLicence: CC BY

Cite this

Pattipeiluhu, R., Arias-Alpizar, G., Basha, G., Chan, K. Y. T., Bussmann, J., Sharp, T. H., Moradi, M. A., Sommerdijk, N., Harris, E. N., Cullis, P. R., Kros, A., Witzigmann, D., & Campbell, F. (2022). Anionic Lipid Nanoparticles Preferentially Deliver mRNA to the Hepatic Reticuloendothelial System. Advanced Materials, 34(16), Article 2201095. https://doi.org/10.1002/adma.202201095

@article{73778fbd78094088ae665fb8268649b6,

title = "Anionic Lipid Nanoparticles Preferentially Deliver mRNA to the Hepatic Reticuloendothelial System",

abstract = "Lipid nanoparticles (LNPs) are the leading nonviral technologies for the delivery of exogenous RNA to target cells in vivo. As systemic delivery platforms, these technologies are exemplified by Onpattro, an approved LNP-based RNA interference therapy, administered intravenously and targeted to parenchymal liver cells. The discovery of systemically administered LNP technologies capable of preferential RNA delivery beyond hepatocytes has, however, proven more challenging. Here, preceded by comprehensive mechanistic understanding of in vivo nanoparticle biodistribution and bodily clearance, an LNP-based messenger RNA (mRNA) delivery platform is rationally designed to preferentially target the hepatic reticuloendothelial system (RES). Evaluated in embryonic zebrafish, validated in mice, and directly compared to LNP{\textendash}mRNA systems based on the lipid composition of Onpattro, RES-targeted LNPs significantly enhance mRNA expression both globally within the liver and specifically within hepatic RES cell types. Hepatic RES targeting requires just a single lipid change within the formulation of Onpattro to switch LNP surface charge from neutral to anionic. This technology not only provides new opportunities to treat liver-specific and systemic diseases in which RES cell types play a key role but, more importantly, exemplifies that rational design of advanced RNA therapies must be preceded by a robust understanding of the dominant nano{\textendash}biointeractions involved.",

keywords = "embryonic zebrafish, lipid nanoparticles, mRNA delivery, reticuloendothelial system, stabilin-2, Mononuclear Phagocyte System/metabolism, RNA, Small Interfering/genetics, Lipids, Zebrafish, RNA, Messenger/metabolism, Nanoparticles, Tissue Distribution, Animals, Liver/metabolism, Mice, Liposomes",

author = "Roy Pattipeiluhu and Gabriela Arias-Alpizar and Genc Basha and Chan, \{Karen Y.T.\} and Jeroen Bussmann and Sharp, \{Thomas H.\} and Moradi, \{Mohammad Amin\} and Nico Sommerdijk and Harris, \{Edward N.\} and Cullis, \{Pieter R.\} and Alexander Kros and Dominik Witzigmann and Frederick Campbell",

note = "Funding Information: R.P. and G.A.‐A. contributed equally to this work. Panagiota Papadopoulou (Leiden University) is thanked for her work in developing apoE‐targeted liposomes within the embryonic zebrafish. Niek Crone (Leiden University) is thanked for gifting 3‐azido‐5‐cholestene. This work was supported financially by the Netherlands Organization for Scientific Research (NWO‐VICI {\textendash} Project No. 724.014.001) {\textendash} R.P., G.A.‐A.; a M‐ERA Grant {\textendash} F.C., M.‐A.M.; the Interreg 2 Seas Program 2014{\textendash}2020 co‐funded by the European Regional Development Fund under subsidy contract {\textquotedblleft}Site Drug 2S07‐033{\textquotedblright} {\textendash} G.A.‐A.; the NanoMedicines Innovation Network (NMIN) {\textendash} G.B., D.W.; and the Swiss National Science Foundation (Grant No. 186741) {\textendash} D.W. Breeding and housing of KO mice was supported by the National Institute of Health (Grant No. HL130864) {\textendash} E.N.H. All mouse protocols were approved by the Canadian Animal Care Committee and conducted in accordance with relevant guidelines and regulations. Zebrafish (Danio rerio, strain AB/TL) were maintained and handled according to the guidelines from the Zebrafish Model Organism Database (http://zfin.org) and in compliance with the directives of the local animal welfare committee of Leiden University. stab2 −/− ",

year = "2022",

month = apr,

day = "21",

doi = "10.1002/adma.202201095",

language = "English",

volume = "34",

journal = "Advanced Materials",

issn = "0935-9648",

publisher = "Wiley-Blackwell",

number = "16",

}

TY - JOUR

T1 - Anionic Lipid Nanoparticles Preferentially Deliver mRNA to the Hepatic Reticuloendothelial System

AU - Pattipeiluhu, Roy

AU - Arias-Alpizar, Gabriela

AU - Basha, Genc

AU - Chan, Karen Y.T.

AU - Bussmann, Jeroen

AU - Sharp, Thomas H.

AU - Moradi, Mohammad Amin

AU - Sommerdijk, Nico

AU - Harris, Edward N.

AU - Cullis, Pieter R.

AU - Kros, Alexander

AU - Witzigmann, Dominik

AU - Campbell, Frederick

N1 - Funding Information: R.P. and G.A.‐A. contributed equally to this work. Panagiota Papadopoulou (Leiden University) is thanked for her work in developing apoE‐targeted liposomes within the embryonic zebrafish. Niek Crone (Leiden University) is thanked for gifting 3‐azido‐5‐cholestene. This work was supported financially by the Netherlands Organization for Scientific Research (NWO‐VICI – Project No. 724.014.001) – R.P., G.A.‐A.; a M‐ERA Grant – F.C., M.‐A.M.; the Interreg 2 Seas Program 2014–2020 co‐funded by the European Regional Development Fund under subsidy contract “Site Drug 2S07‐033” – G.A.‐A.; the NanoMedicines Innovation Network (NMIN) – G.B., D.W.; and the Swiss National Science Foundation (Grant No. 186741) – D.W. Breeding and housing of KO mice was supported by the National Institute of Health (Grant No. HL130864) – E.N.H. All mouse protocols were approved by the Canadian Animal Care Committee and conducted in accordance with relevant guidelines and regulations. Zebrafish (Danio rerio, strain AB/TL) were maintained and handled according to the guidelines from the Zebrafish Model Organism Database (http://zfin.org) and in compliance with the directives of the local animal welfare committee of Leiden University. stab2 −/−

PY - 2022/4/21

Y1 - 2022/4/21

N2 - Lipid nanoparticles (LNPs) are the leading nonviral technologies for the delivery of exogenous RNA to target cells in vivo. As systemic delivery platforms, these technologies are exemplified by Onpattro, an approved LNP-based RNA interference therapy, administered intravenously and targeted to parenchymal liver cells. The discovery of systemically administered LNP technologies capable of preferential RNA delivery beyond hepatocytes has, however, proven more challenging. Here, preceded by comprehensive mechanistic understanding of in vivo nanoparticle biodistribution and bodily clearance, an LNP-based messenger RNA (mRNA) delivery platform is rationally designed to preferentially target the hepatic reticuloendothelial system (RES). Evaluated in embryonic zebrafish, validated in mice, and directly compared to LNP–mRNA systems based on the lipid composition of Onpattro, RES-targeted LNPs significantly enhance mRNA expression both globally within the liver and specifically within hepatic RES cell types. Hepatic RES targeting requires just a single lipid change within the formulation of Onpattro to switch LNP surface charge from neutral to anionic. This technology not only provides new opportunities to treat liver-specific and systemic diseases in which RES cell types play a key role but, more importantly, exemplifies that rational design of advanced RNA therapies must be preceded by a robust understanding of the dominant nano–biointeractions involved.

AB - Lipid nanoparticles (LNPs) are the leading nonviral technologies for the delivery of exogenous RNA to target cells in vivo. As systemic delivery platforms, these technologies are exemplified by Onpattro, an approved LNP-based RNA interference therapy, administered intravenously and targeted to parenchymal liver cells. The discovery of systemically administered LNP technologies capable of preferential RNA delivery beyond hepatocytes has, however, proven more challenging. Here, preceded by comprehensive mechanistic understanding of in vivo nanoparticle biodistribution and bodily clearance, an LNP-based messenger RNA (mRNA) delivery platform is rationally designed to preferentially target the hepatic reticuloendothelial system (RES). Evaluated in embryonic zebrafish, validated in mice, and directly compared to LNP–mRNA systems based on the lipid composition of Onpattro, RES-targeted LNPs significantly enhance mRNA expression both globally within the liver and specifically within hepatic RES cell types. Hepatic RES targeting requires just a single lipid change within the formulation of Onpattro to switch LNP surface charge from neutral to anionic. This technology not only provides new opportunities to treat liver-specific and systemic diseases in which RES cell types play a key role but, more importantly, exemplifies that rational design of advanced RNA therapies must be preceded by a robust understanding of the dominant nano–biointeractions involved.

KW - embryonic zebrafish

KW - lipid nanoparticles

KW - mRNA delivery

KW - reticuloendothelial system

KW - stabilin-2

KW - Mononuclear Phagocyte System/metabolism

KW - RNA, Small Interfering/genetics

KW - Lipids

KW - Zebrafish

KW - RNA, Messenger/metabolism

KW - Nanoparticles

KW - Tissue Distribution

KW - Animals

KW - Liver/metabolism

KW - Mice

KW - Liposomes

UR - https://www.scopus.com/pages/publications/85125992936

U2 - 10.1002/adma.202201095

DO - 10.1002/adma.202201095

M3 - Article

C2 - 35218106

AN - SCOPUS:85125992936

SN - 0935-9648

VL - 34

JO - Advanced Materials

JF - Advanced Materials

IS - 16

M1 - 2201095

ER -

Anionic Lipid Nanoparticles Preferentially Deliver mRNA to the Hepatic Reticuloendothelial System

Abstract

Bibliographical note

Funding

Keywords

Access to Document

Other files and links

Fingerprint

Cite this