Naturally-occurring cholesterol analogues in lipid nanoparticles induce polymorphic shape and enhance intracellular delivery of mRNA
Open accessAbstract
Endosomal sequestration of lipid-based nanoparticles (LNPs) remains a formidable barrier to delivery. Herein, structure-activity analysis of cholesterol analogues reveals that incorporation of C-24 alkyl phytosterols into LNPs (eLNPs) enhances gene transfection and the length of alkyl tail, flexibility of sterol ring and polarity due to -OH group is required to maintain high transfection. Cryo-TEM displays a polyhedral shape for eLNPs compared to spherical LNPs, while x-ray scattering shows little disparity in internal structure. eLNPs exhibit higher cellular uptake and retention, potentially leading to a steady release from the endosomes over time. 3D single-particle tracking shows enhanced intracellular diffusivity of eLNPs relative to LNPs, suggesting eLNP traffic to productive pathways for escape. Our findings show the importance of cholesterol in subcellular transport of LNPs carrying mRNA and emphasize the need for greater insights into surface composition and structural properties of nanoparticles, and their subcellular interactions which enable designs to improve endosomal escape.
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In mRNA-LNP formulations, the fraction of accessible mRNA is very low and encapsulation efficiencies are typically > 90%.
“In mRNA-LNP formulations, such as those used in mRNA vaccines, the fraction of accessible mRNA is very low (Yanez Arteta et al., 2018, Patel et al., 2020) and thus, encapsulation efficiencies, derived from RiboGreen assays, are typically > 90%.”
From mRNA-lipid nanoparticle COVID-19 vaccines: structure and stability (2021) · cites this paper as Patel et al., 2020
Other researchers propose that the outer shell of mRNA-LNPs consists of one or multiple bilayers.
“Other researchers propose that the outer shell of mRNA-LNPs consists of one or multiple bilayers, based on cryo-TEM analysis (Fig. 2) (Patel et al., 2020, Tanaka et al., 2020) or SANS (Sebastiani et al., 2021).”
From mRNA-lipid nanoparticle COVID-19 vaccines: structure and stability (2021) · cites this paper as Patel et al., 2020
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- Authors
- Patel, S., Ashwanikumar, N., Robinson, E., Xia, Y., Mihai, C., Griffith, J. P., Hou, S., Esposito, A. A., Ketova, T., Welsher, K., Joyal, J. L., Almarsson, Ö., & Sahay, G.
- Affiliations
- Oregon State University (US)
- Moderna Therapeutics (United States) (US)
- Duke University (US)
- Corresponding author
- Sahay, Gaurav
- ORCID
- Venue
- Nature Communications
- Published
- 2020-02-20
- Volume
- 11
- Issue
- 1
- Pages
- 983-983
- ISSN
- 2041-1723 (ISSN-L)
- PMC
- PMC7033178
- OpenAlex
- W3006764150
- Cited by
- 601
- References
- 72
- Citations by year
- 2026: 108 · 2025: 156 · 2024: 127 · 2023: 72 · 2022: 64 · 2021: 50 · 2020: 23
- FWCI
- 16.94
- Type
- journal article
- Language
- en
- Update notices
- correction (2020-07-06)
- Open access
- gold
- doi.org
- Locations
- Nature Communications (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- U.S. Department of Health & Human Services | NIH | National Institute of Biomedical Imaging and Bioengineering (1R15EB021581)
- U.S. Department of Health & Human Services | NIH | National Heart, Lung, and Blood Institute (1R01HL146736-01)
- Cystic Fibrosis Foundation (SAHAY18G0)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmaceutical Science › Advancements in Transdermal Drug Delivery
- Keywords
- cholesterol analogues, lipid nanoparticles, mRNA delivery, endosomal escape, nanoparticle morphology, transfection, cellular uptake, intracellular diffusion, structure-activity relationship
- MeSH terms
- RAW 264.7 Cells, Animals, Biological Transport, Active, Cell Line, Cholesterol, HeLa Cells, Humans, Lipids, Endosomes, RNA, Messenger, Sitosterols, Transfection, X-Ray Diffraction, Cryoelectron Microscopy, Mice, Nanoparticles, HEK293 Cells
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