Interaction of non-lamellar liquid-crystalline lipid carpets with model endosomal vesicles.

Nucleic acid (NA) therapeutics hold great potential, but their success relies on delivery systems, able to promote effective endosomal
escape. Emerging evidence suggests that non-lamellar lipid nanoparticles (NLLNPs), with highly curved bilayers, may enhance
endosomal escape. We have developed a microfluidic technique to produce NLLNPs using glycerol monooleate and ionizable lipids,
which combine topological activity with pH sensitivity. Preliminary studies show these NLLNPs effectively bind to and destabilize
endosomal membranes under acidic conditions, but the mechanism remains unclear. We propose to use Neutron Reflectometry (NR) to
investigate the mechanism by which NLLNPs destabilize endosomal membranes as a function of pH. To this end, we will employ
innovative solid-supported nanometric “carpets” of NLLNPs and study their interactions with model endosomal vesicles in real-time
using Time-Resolved NR. Leveraging selective deuteration and contrast variation, we aim to resolve structural changes and lipid
exchange processes occurring during vesicle binding and fusion with NLLNPs. These insights will inform the design of more efficient
lipid nanoparticles for NA delivery.