Protein Corona Engineering of Lipid Nanoparticles Enables Efficient Functional Gene Silencing in the Liver

ABSTRACT Lipid nanoparticles (LNPs) preferentially accumulate in the liver, but hepatic tropism alone does not ensure productive RNA interference. Here, we screened a library of LNPs in which lipid composition was varied to generate distinct protein corona identities and identify formulations capable of combining liver accumulation with functional siRNA delivery. LNP1 emerged as the lead formulation, showing favorable physicochemical properties, high transfection efficiency, and enhanced hepatic accumulation. Proteomic analysis associated this behavior with a distinct multicomponent corona signature. When loaded with siRNA targeting tissue inhibitor of metalloproteinases 1 (TIMP1), a key regulator of extracellular matrix turnover and fibrosis, LNP1 was efficiently internalized by primary Kupffer cells (KCs) and induced robust target knockdown. Consistent with this cellular preference, quartz crystal microbalance measurements showed that corona formation increased LNP1 binding to MARCO, a scavenger receptor expressed by KCs. In a mouse model of primary biliary cholangitis, repeated administration of siTIMP1-loaded LNP1 reduced TIMP1 expression in KCs, reactivated extracellular-matrix-degradative pathways, suppressed pro-inflammatory signaling and macrophage infiltration, alleviated cholestasis and attenuated liver fibrosis. These findings connect systematic variation in lipid composition with corona identity, hepatic tropism, and functional gene silencing, supporting protein corona engineering as a strategy to bridge liver accumulation with productive siRNA delivery.