A ternary nanocomplex for hepatocyte-targeted gene delivery and treatment of metabolic dysfunction-associated steatotic liver disease.

The liver, with its unique physiology and powerful metabolic activity, represents an ideal 'protein factory' for gene therapy. Yet efficient, cell-specific transfection of hepatocytes remains a significant challenge. To address this, we engineered a novel ternary nanocomplex through a modular design. A stable binary core was first formed by electrostatically condensing plasmid DNA (pDNA) with a fluorinated ROS-cleavable (TK) polyethylenimine (FRP), enhancing cellular uptake and facilitating endosomal escape. This core was subsequently coated with a surface layer of galactose-modified hyaluronic acid-polyethylene glycol (GPH), enabling dual-receptor-mediated hepatocyte targeting. Galactose (Gal) engages the asialoglycoprotein receptor (ASGPR) and hyaluronic acid (HA) interacts with CD44 on hepatocytes, while the PEG corona improves complex stability. In vitro studies demonstrated that the FRP/pDNA and GP (GPH-coated FRP/pDNA) complexes achieved highly efficient and specific gene transfection in AML12 hepatocytes, significantly outperforming the commercial reagent Lipofectamine 3000. In a cellular model of metabolic dysfunction-associated steatotic liver disease (MASLD), successful FGF21 gene delivery via FRP or GP effectively alleviated lipid accumulation. This work highlights the potential of the GP nanocomplex as a powerful platform for hepatocyte-targeted gene delivery, offering a promising strategy for treating liver-centric metabolic disorders.