Targeting Amphotericin B Delivery to Yeast with ApoA1 Lipid Nanodiscs Coupled to Dectin-1 Using a Modular SpyCatcher–SpyTag System

Lipid nanodiscs are synthetic nanoparticles capable of solubilizing lipophilic drugs and have been shown to improve the potency of the antifungal Amphotericin B (AmphB) against various fungal pathogens. In this study, the SpyCatcher–SpyTag covalent labeling system was used to couple AmphB-loaded Apolipoprotein A1 (ApoA1) lipid nanodiscs to the receptor domain of Dectin-1, which binds to β-1,3/1,6 glucans present in many fungal cell walls. Denaturing protein gel electrophoresis demonstrated that ApoA1-SpyTag003 lipid nanodiscs could be covalently labeled with SpyCatcher003-Dectin-1-superfolder GFP (sfGFP). In microtiter growth assays with Saccharomyces cerevisiae, Dectin-1 AmphB nanodiscs displayed an IC50 1.5-fold lower than uncoupled AmphB nanodiscs and 2.8-fold lower than AmphB-only controls. Nanodiscs without AmphB and SpyCatcher003-Dectin-1-sfGFP themselves did not inhibit yeast growth. Fluorescence microscopy showed that SpyCatcher003-Dectin-1-sfGFP binds to yeast cell walls and accumulated at hot spots, matching the budding scar enrichment pattern previously described for other Dectin-1 fusion proteins. Together these results indicate that Dectin-1 fusions can target AmphB-loaded lipid nanodiscs to fungal cell walls and improve drug delivery. The results here establish the use of a modular SpyCatcher–SpyTag coupling system for targeting drug-loaded lipid nanodiscs to different cells or tissues, thereby increasing drug retention at infection sites, increasing drug potency, and reducing harmful side-effects.