Cited by 48RNA Interference and Gene DeliveryAdvancements in Transdermal Drug DeliveryNanoparticle-Based Drug Delivery

Effect of complexing lipids on cellular uptake and expression of messenger RNA in human skin explants

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Blakney, Anna K., Deletic, Polina, McKay, Paul F., Bouton, Clément R., Ashford, Marianne, Shattock, Robin J., Sabirsh, Alan·2021·Journal of Controlled Release

Abstract

Messenger RNA (mRNA) represents a promising next-generation approach for both treatment and vaccination. Lipid based particles are one of the most investigated delivery systems for mRNA formulations. Here we explore how the complexing lipid affects uptake and translation of lipoplex-delivered RNA in resident cells in human skin explants and, we explore a more modular delivery system that utilizes mRNA added to pre-formed nanoparticles prior to dosing. We prepared formulations of lipoplexes with ionizable, cationic or zwitterionic lipids, externally complexed these with mRNA, and observed which cells internalized and/or expressed the mRNA over 72 h after intradermal injections into primary, human, skin explants. Using a flow cytometry panel to assess cellular phenotypes, mRNA uptake and mRNA expression, we found that, unlike other cell types, adipocytes expressed mRNA efficiently at 4 and 24 h after mRNA-lipoplex injection and contributed the greatest proportion of total RNA-encoded protein expression, despite being the lowest frequency cell type. Other cell types (epithelial cells, fibroblasts, T cells, B cells, dendritic cells, monocytes, NK cells, Langerhans cells, and leukocytes) had increasing mRNA expression over the course of 72 h, irrespective of lipoplex formulation. We observed that overall charge of the particle, but not the complexing lipid classification, was predictive for the pattern of mRNA uptake and expression among resident cell types in this model. This study provides insight into maximizing protein expression, using modular mRNA lipoplexes that are more compatible with product development, in a clinically relevant, human skin explant model.

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Crossref · 10.1016/j.jconrel.2020.11.033

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Authors
Blakney, A. K., Deletic, P., McKay, P. F., Bouton, C. R., Ashford, M., Shattock, R. J., & Sabirsh, A.
Affiliations
  • Imperial College London (GB)
  • AstraZeneca (United Kingdom) (GB)
  • AstraZeneca (Sweden) (SE)
Corresponding author
Blakney, Anna K.
Venue
Journal of Controlled Release
Publisher
Elsevier BV
Published
2021-02
Volume
330
Pages
1250-1261
ISSN
0168-3659 (ISSN-L)
OpenAlex
W3109389550
Cited by
48
References
41
Citations by year
2026: 7 · 2025: 4 · 2024: 10 · 2023: 6 · 2022: 10 · 2021: 11
FWCI
2.19
Type
journal article
Language
en
Open access
Locations
  • Journal of Controlled Release (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • Spiral (Imperial College London) (submittedVersion, cc-by-nc-nd)
Funders
  • Engineering and Physical Sciences Research Council
  • H2020 Marie Skłodowska-Curie Actions
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmaceutical Science › Advancements in Transdermal Drug Delivery
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
Keywords
mRNA delivery, lipoplexes, ionizable lipids, cationic lipids, zwitterionic lipids, skin explants, intradermal injection, mRNA expression, adipocytes, particle charge
MeSH terms
Cations, Humans, Lipids, Liposomes, RNA, Messenger, Skin, Transfection, Nanoparticles

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