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Nanoparticle stereochemistry-dependent endocytic processing improves in vivo mRNA delivery
Marine Z C Hatit1, Curtis N Dobrowolski1, Melissa P Lokugamage1
1Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, GA, USA.
Nature Chemistry
|March 2, 2023
Summary
Lipid nanoparticle (LNP) stereochemistry impacts mRNA delivery potency. Stereopure hydroxycholesterol LNPs show superior liver cell delivery compared to mixed forms, highlighting the role of molecular structure in LNP efficacy.
Area of Science:
- Biochemistry
- Nanomedicine
- Molecular Biology
Background:
- Stereochemistry significantly influences the pharmacokinetic, safety, and efficacy profiles of small-molecule drugs.
- The impact of stereochemistry within multicomponent colloidal systems, like lipid nanoparticles (LNPs), on in vivo activity remains largely unexplored.
Purpose of the Study:
- To investigate whether the stereochemistry of a specific compound within a lipid nanoparticle (LNP) affects its in vivo mRNA delivery efficiency.
- To elucidate the mechanisms underlying any observed stereochemistry-dependent differences in LNP activity.
Main Methods:
- Lipid nanoparticles (LNPs) were formulated with stereopure 20α-hydroxycholesterol (20α) or a mixture of 20α- and 20β-hydroxycholesterols (20mix).
- In vivo potency was assessed by measuring mRNA delivery to liver cells.
- Single-cell RNA sequencing and imaging were employed to analyze LNP biodistribution and cellular interactions.
- Physicochemical properties of the LNPs were evaluated.
Main Results:
- LNPs containing stereopure 20α-hydroxycholesterol demonstrated up to a 3-fold increase in mRNA delivery potency to liver cells compared to LNPs with mixed hydroxycholesterols.
- This enhanced delivery was not attributable to differences in LNP physicochemical characteristics.
- In vivo analysis revealed that mixed hydroxycholesterol LNPs were preferentially directed into phagocytic pathways, leading to distinct biodistribution and functional delivery outcomes.
- Stereochemistry-dependent cellular interactions were identified as a key factor influencing LNP efficacy.
Conclusions:
- The stereochemistry of hydroxycholesterol within LNPs significantly modulates mRNA delivery efficiency in vivo.
- Stereopure 20α-hydroxycholesterol-containing LNPs enhance mRNA delivery potency by altering cellular uptake and trafficking pathways.
- These findings underscore the importance of stereochemical control in LNP design for improved nanomedicine applications.
