Phase-Separated Lipid-Based Nanoparticles: Selective Behavior at the Nano-Bio Interface

The membrane-protein interface on lipid-based nanoparticles influences their in vivo behavior. Better understanding may evolve current drug delivery methods toward effective targeted nanomedicine. Previously, the cell-selective accumulation of a liposome formulation in vivo is demonstrated, through the recognition of lipid phase-separation by triglyceride lipases. This exemplified how liposome morphology and composition can determine nanoparticle-protein interactions. Here, the lipase-induced compositional and morphological changes of phase-separated liposomes-which bear a lipid droplet in their bilayer- are investigated, and the mechanism upon which lipases recognize and bind to the particles is unravelled. The selective lipolytic degradation of the phase-separated lipid droplet is observed, while nanoparticle integrity remains intact. Next, the Tryptophan-rich loop of the lipase is identified as the region with which the enzymes bind to the particles. This preferential binding is due to lipid packing defects induced on the liposome surface by phase separation. In parallel, the existing knowledge that phase separation leads to in vivo selectivity, is utilized to generate phase-separated mRNA-LNPs that target cell-subsets in zebrafish embryos, with subsequent mRNA delivery and protein expression. Together, these findings can expand the current knowledge on selective nanoparticle-protein communications and in vivo behavior, aspects that will assist to gain control of lipid-based nanoparticles. The synthetic diacylglycerol analogue lipid DOaG induces phase separation in lipid-based nanoparticles, generating distinct lipid droplets. In liposomes, DOaG-induced lipid packing defects facilitate recognition and recruitment of lipoprotein lipase (LPL), with subsequent selective hydrolysis of the DOaG-rich lipid droplet. In mRNA-LNPs DOaG induces phase separation, leading to selective cell targeting, mRNA delivery, and concomitant protein expression in vivo.image