ISIS Neutron and Muon Source Data Journal

Characterising nanoscale structural determinants of effective intranasal mRNA delivery

Abstract: Intramuscular mRNA vaccines fail to elicit immunity in the respiratory tract, underscoring the need for delivery systems capable of local transfection and immunogenicity in mucosal tissues. Positively charged polymer- and lipid-based nanoparticles demonstrate superior intranasal mRNA delivery, yet the nanoscale structural determinants governing this efficacy remain poorly defined. We propose to use small-angle neutron scattering (SANS) to directly probe how helper lipid composition and cationic charge influence the multiscale structure of lipid nanoparticles (LNPs) and polylipidoid nanoparticles (pLNPs). Preliminary SAXS, SANS, and cryo-EM analyses reveal that the cationic lipid DOTAP tightens RNA packaging in pLNPs and LNPs but with opposing impacts on particle size and mesoscale ordering. We hypothesize that mesoscale cationic lipid ordering affects glycosaminoglycan binding and the mucosal transfection of these formulations. Leveraging deuterated helper lipids and solvent contrast matching, we will use SANS to quantify lipid ordering, mRNA-lipid phase separation, and the structural remodeling of nanoparticles upon interaction with mucosal glycosaminoglycans. SANS uniquely enables in situ measurements at therapeutically relevant concentrations. This first SANS characterization of pLNPs, will define structure-function relationships underlying mucosal transfection and guide design of intranasal mRNA vaccines and therapeutics.

Principal Investigator: Mr Vincent Kruit
Experimenter: Dr Xinyu Lu
Experimenter: Dr Massimo Francesco Cau
Experimenter: Professor Molly Stevens
Local Contact: Dr James Doutch
Experimenter: Dr David Peeler
Experimenter: Dr Anna Constantinou
Experimenter: Dr Shahd Qassem
Experimenter: Dr Patrick Peschke
Experimenter: Mr Kevin Alejandro Giraldo Rodriguez

DOI: 10.5286/ISIS.E.RB2610658

ISIS Experiment Number: RB2610658

Publisher: STFC ISIS Neutron and Muon Source

Data format: RAW/Nexus
Select the data format above to find out more about it.

Data Citation

The recommended format for citing this dataset in a research publication is as:
[author], [date], [title], [publisher], [doi]

For Example:
Mr Vincent Kruit et al; (2026): Characterising nanoscale structural determinants of effective intranasal mRNA delivery, STFC ISIS Neutron and Muon Source, https://doi.org/10.5286/ISIS.E.RB2610658

Data is released under the CC-BY-4.0 license.