Interaction Kinetics of Individual mRNA-Containing Lipid Nanoparticles with an Endosomal Membrane Mimic: Dependence on pH, Protein Corona Formation, and Lipoprotein Depletion
Journal article, 2022

Lipid nanoparticles (LNPs) have emerged as potent carriers for mRNA delivery, but several challenges remain before this approach can offer broad clinical translation of mRNA therapeutics. To improve their efficacy, a better understanding is required regarding how LNPs are trapped and processed at the anionic endosomal membrane prior to mRNA release. We used surface-sensitive fluorescence microscopy with single LNP resolution to investigate the pH dependency of the binding kinetics of ionizable lipid-containing LNPs to a supported endosomal model membrane. A sharp increase of LNP binding was observed when the pH was lowered from 6 to 5, accompanied by stepwise large-scale LNP disintegration. For LNPs preincubated in serum, protein corona formation shifted the onset of LNP binding and subsequent disintegration to lower pH, an effect that was less pronounced for lipoprotein-depleted serum. The LNP binding to the endosomal membrane mimic was observed to eventually become severely limited by suppression of the driving force for the formation of multivalent bonds during LNP attachment or, more specifically, by charge neutralization of anionic lipids in the model membrane due to their association with cationic lipids from earlier attached LNPs upon their disintegration. Cell uptake experiments demonstrated marginal differences in LNP uptake in untreated and lipoprotein-depleted serum, whereas lipoprotein-depleted serum increased mRNA-controlled protein (eGFP) production substantially. This complies with model membrane data and suggests that protein corona formation on the surface of the LNPs influences the nature of the interaction between LNPs and endosomal membranes.

mRNA delivery

protein corona

ionizable lipid nanoparticle

lipoprotein

endosomal membrane

Author

Nima Aliakbarinodehi

Chalmers, Physics, Nano and Biophysics

Audrey Gallud

Chalmers, Biology and Biological Engineering, Chemical Biology

AstraZeneca AB

Mokhtar Mapar

Chalmers, Physics, Nano and Biophysics

Emelie Vilhelmsson Wesén

Chalmers, Biology and Biological Engineering, Chemical Biology

Sahar Heydari

Chalmers, Biology and Biological Engineering, Chemical Biology

Yujia Jing

AstraZeneca AB

Gustav Emilsson

AstraZeneca AB

Kai Liu

AstraZeneca AB

Alan Sabirsh

AstraZeneca AB

Vladimir Zhdanov

Russian Academy of Sciences

Chalmers, Physics

Lennart Lindfors

AstraZeneca AB

Elin Esbjörner Winters

Chalmers, Biology and Biological Engineering, Chemical Biology

Fredrik Höök

Chalmers, Physics, Nano and Biophysics

ACS Nano

1936-0851 (ISSN) 1936-086X (eISSN)

Vol. 16 12 20163-20173

Subject Categories

Physical Chemistry

Biochemistry and Molecular Biology

Biophysics

DOI

10.1021/acsnano.2c04829

PubMed

36511601

More information

Latest update

3/7/2024 9