Sequential Precise Targeted Hepatitis B Virus Therapy: Hepatitis B Virus-Specific Small Interfering RNA Followed by Aggregation-Induced Emission-Photothermal Ablation Based on Macrophage Membrane-Coated Nanoplatform.

Chronic hepatitis B (CHB) afflicts approximately 254 million people worldwide and is responsible for more than one million deaths annually. Intrahepatic persistence of covalently closed circular DNA (cccDNA), the episomal transcriptional template of hepatitis B virus (HBV), sustains chronic replication, blunts the efficacy of antiviral therapy, and precipitates virological relapse once treatment is discontinued. While emerging cccDNA-targeted strategies include CRISPR/Cas9-mediated cleavage and epigenetic silencing, their clinical translation is impeded by delivery inefficiency, safety concerns, and limited durability. Herein, we present a sequential nano-therapeutic platform that combines small interfering RNA (siRNA) delivery using HBV-pre-activated macrophage membranes (HMs) with photothermal therapy utilizing aggregation-induced emission (AIE) agents. In the first step, HM-coated lipid nanoparticles encapsulating siRNA (siR@HM NPs) enable virus to neutralize and hepatocyte targeted delivery via surface viral receptors, achieving efficient gene silencing. At 72 h post-administration, HM-coated PLGA nanoparticles loaded with an AIE photothermal agent (TPAT@HM NPs) are introduced, facilitating near-infrared IIb fluorescence-guided 808 nm laser ablation of residual infected cells. In a cell model of chronic HBV infection, both siR@HM NPs and TPAT@HM NPs achieved targeted delivery to infected hepatocytes. Critically, sequential administration yielded superior antiviral efficacy compared with concurrent administration or either monotherapy, leading to substantial reduction in HBsAg, HBeAg, HBV DNA, cccDNA, and pgRNA levels. Biosafety assessments confirmed negligible cytotoxicity, hemolysis, and systemic toxicity. This bionic platform integrates time-programmed virus targeting, RNA interference, and photothermal ablation. The virus-preactivated membrane coating enables dual recognition of both viral particles and infected cells, offering a precise and scalable theranostic strategy for HBV and other refractory viral infections.