The Skin You Are In: Design-of-Experiments Optimization of Lipid Nanoparticle Self-Amplifying RNA Formulations in Human Skin Explants
Open accessAbstract
Messenger RNA (mRNA) is a promising tool for biotherapeutics, and self-amplifying mRNA (saRNA) is particularly advantageous, because it results in abundant protein expression and production is easily scalable. While mRNA therapeutics have been shown to be highly effective in small animals, the outcomes do not scale linearly when these formulations are translated to dose-escalation studies in humans. Here, we utilize a design of experiments (DoE) approach to optimize the formulation of saRNA lipid nanoparticles in human skin explants. We first observed that luciferase expression from saRNA peaked after 11 days in human skin. Using DoE inputs of complexing lipid identity, lipid nanoparticle dose, lipid concentration, particle concentration, and ratio of zwitterionic to cationic lipids, we optimized the saRNA-induced luciferase expression in skin explants. Lipid identity and lipid concentration were found to be significant parameters in the DoE model, and the optimized formulation resulted in ∼7-fold increase in luciferase expression, relative to initial 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) formulation. Using flow cytometry, we observed that optimized formulations delivered the saRNA to ∼2% of the resident cells in the human skin explants. Although immune cells comprise only 7% of the total population of cells in skin, immune cells were found to express ∼50% of the RNA. This study demonstrates the powerful combination of using a DoE approach paired with clinically relevant human skin explants to optimize nucleic acid formulations. We expect that this system will be useful for optimizing both formulation and molecular designs of clinically translational nucleic acid vaccines and therapeutics.
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mRNA-based protein replacement therapies have been applied to haematological diseases, central nervous system disorders, skin diseases and hearing loss in preclinical studies.
“In addition, mRNA-based protein replacement therapies have been applied to haematological diseases (for example, haemophilia A232, haemophilia B91,104,233 and thrombotic thrombocytopaenic purpura234), central nervous system disorders67,152 (for example, Friedreich’s ataxia67), skin diseases235,236 (for example, elastin deficiency235) and hearing loss237 in preclinical”
From Lipid nanoparticles for mRNA delivery (2021) · cites this paper as [236]
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- Authors
- Blakney, A. K., McKay, P. F., Ibarzo Yus, B., Hunter, J. E., Dex, E. A., & Shattock, R. J.
- Affiliations
- Imperial College London (GB)
- Imperial College London , , , W21PG ,
- Imperial NHS Trust , , , ,
- Corresponding author
- Shattock, Robin J.
- Venue
- ACS Nano
- Publisher
- American Chemical Society (ACS)
- Published
- 2019-05-02
- Volume
- 13
- Issue
- 5
- Pages
- 5920-5930
- ISSN
- 1936-0851 (ISSN-L)
- 1936-086X
- PMC
- PMC7007275
- OpenAlex
- W2943649234
- Cited by
- 73
- References
- 38
- Citations by year
- 2026: 9 · 2025: 10 · 2024: 12 · 2023: 12 · 2022: 6 · 2021: 12 · 2020: 11 · 2019: 1
- FWCI
- 2.47
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- ACS Nano (publishedVersion, cc-by)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Spiral (Imperial College London) (submittedVersion, cc-by)
- Funders
- Whitaker International Fellows and Scholars Program (NA)
- Engineering and Physical Sciences Research Council (NA)
- Innovate UK (NA)
- Imperial Confidence in Concept (RSRO P75207)
- 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
- self-amplifying mRNA, lipid nanoparticles, human skin explants, experimental design, luciferase expression, DOTAP, nucleic acid delivery
- MeSH terms
- Animals, Humans, Lipids, Liposomes, RNA, RNA, Messenger, Skin, Transfection, Nanoparticles
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