lncRNA-driven multi-omics reconstitution engineers EVs for macrophage-targeted and anti-inflammatory therapy in MASLD.

Extracellular vesicles (EVs) can encapsulate and deliver diverse bioactive cargos, enabling multimolecular parallel communication and network-level regulation, and are emerging as promising nanotherapeutics. However, existing engineering strategies predominantly focus on single-function enhancement, limiting coordinated improvements in multi-dimensional regulation. As a proof of concept, we propose a lncRNA-driven multi-omics (transcriptomic and proteomic) reconstitution EV engineering strategy that leverages synergistic functional enhancement to generate multifunctional EVs (MF-EVs) with hepatic macrophage targeting and anti-inflammatory activity for the treatment of metabolic dysfunction-associated steatotic liver disease (MASLD). Using HEK-293T as a chassis cells, overexpression of the lncENAF reprogrammed cellular states and increased EV yield by 40%. Proteomic and miRNA sequencing analyses revealed that the resulting MF-EVs are enriched for key molecular networks involved in inflammatory regulation and lipid metabolic homeostasis. Evidence from in vitro and in vivo studies demonstrated that MF-EVs exhibit efficient, selective delivery to hepatic macrophages, exert potent anti-inflammatory effects, and reduce the release of pro-inflammatory mediators. Functionally, MF-EVs significantly ameliorated hepatic inflammation and histopathological injury in high-fat diet-induced MASLD mice. Departing from conventional single-point enhancement, our approach treats lncRNA as a systems-level input to achieve endogenous, multidimensional reconfiguration of producer cells and their EVs, establishing an engineering route and conceptual basis for EV therapies targeting complex network diseases.