Apolipoprotein E Binding Drives Structural and Compositional Rearrangement of mRNA-Containing Lipid Nanoparticles
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Emerging therapeutic treatments based on the production of proteins by delivering mRNA have become increasingly important in recent times. While lipid nanoparticles (LNPs) are approved vehicles for small interfering RNA delivery, there are still challenges to use this formulation for mRNA delivery. LNPs are typically a mixture of a cationic lipid, distearoylphosphatidylcholine (DSPC), cholesterol, and a PEG-lipid. The structural characterization of mRNA-containing LNPs (mRNA-LNPs) is crucial for a full understanding of the way in which they function, but this information alone is not enough to predict their fate upon entering the bloodstream. The biodistribution and cellular uptake of LNPs are affected by their surface composition as well as by the extracellular proteins present at the site of LNP administration, e.g., apolipoproteinE (ApoE). ApoE, being responsible for fat transport in the body, plays a key role in the LNP's plasma circulation time. In this work, we use small-angle neutron scattering, together with selective lipid, cholesterol, and solvent deuteration, to elucidate the structure of the LNP and the distribution of the lipid components in the absence and the presence of ApoE. While DSPC and cholesterol are found to be enriched at the surface of the LNPs in buffer, binding of ApoE induces a redistribution of the lipids at the shell and the core, which also impacts the LNP internal structure, causing release of mRNA. The rearrangement of LNP components upon ApoE incubation is discussed in terms of potential relevance to LNP endosomal escape.
| Original language | English |
|---|---|
| Journal | ACS Nano |
| Volume | 15 |
| Issue number | 4 |
| Pages (from-to) | 6709-6722 |
| Number of pages | 14 |
| ISSN | 1936-0851 |
| DOIs | |
| Publication status | Published - 2021 |
| Externally published | Yes |
Bibliographical note
Funding Information:
F.S. acknowledges support from the Knowledge Foundation (Sweden) with a ProSpekt grant (20180101). M.C. thanks the Swedish Research Council for financial support (2014-3981, 2018-03990, and 2018-0483). The authors thank the ILL (Grenoble, France) for allocations of beam time on D22 with a corresponding DOI number: 10.5291/ILL-DATA.9-13-866. This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement no. 731019 (EUSMI) to access beamtime at the KWS-2 instrument operated by JCNS at Heinz Maier-Leibnitz Zentrum, Garching, Germany. We thank Aurel Radulescu for help with data reduction and instrument configuration at KWS-2 and Christopher Garvey for discussions during the beamtime. V.T.F. acknowledges the UK Engineering and Physical Sciences Research Council (EPSRC) for grants EP/C015452/1 and GR/R99393/01 under which the Deuteration Laboratory within ILL’s Life Sciences Group was created. We thank Gernot A. Strohmeier for purifying the deuterated cholesterol (average 89% D). We also thank Linda Thunberg for purifying the deuterated MC3. Yeast strain RH6829 for deuterated cholesterol was kindly provided by Howard Riezman (University of Geneva, Switzerland). The National Deuteration Facility in Australia is partly funded by The National Collaborative Research Infrastructure Strategy (NCRIS), an Australian Government initiative. This work benefited from the use of the SasView application, originally developed under NSF award DMR-0520547. SasView contains code developed with funding from the European Union’s Horizon 2020 research and innovation program under the SINE2020 project, grant agreement no. 654000.
Publisher Copyright:
© 2021 American Chemical Society.
- ApoE, lipid nanoparticles, mRNA delivery, protein corona, small-angle scattering
Research areas
ID: 348241667