Branched ligand-functionalized immuno-evasive PEGylation (BLIP) platform: Breaking through accelerated blood clearance and chemoresistance barriers with dual-targeting RNAi-chemotherapy synergy

PEGylation enhances nanoparticle stability and biocompatibility, but anti-PEG antibodies limit the clinical translation of linear PEGylation. A novel branched ligand-functionalized immuno-evasive PEGylation (BLIP) platform based on lipid nanoparticles (LNP) is developed in this study to break through the immunogenicity barrier of linear PEGylation and evaluated in a chemoresistant non-small cell lung cancer (NSCLC) model through an RNA interference (RNAi)-chemotherapy synergy strategy. Branched ligand-functionalized PEG derivative (AEAA-PEG 2,2k -DSPE) is synthesized to construct the BLIP platform by modifying LNPs, which subsequently co-load Gli1-siRNA (siGli1) and paclitaxel (PTX) to form APL B -siGli1/PTX. Compared to the linear counterpart APL L , the branched PEG modification achieves prolonged blood circulation and stable pharmacokinetics upon repeated injection (AUC APLB = 3.6 × AUC APLL ), avoiding the accelerated blood clearance (ABC) phenomenon. The introduction of targeting ligand enables dual targeting of PTX-resistant NSCLC cells (A549/PTX) and cancer-associated fibroblasts (CAF), enhancing nanoparticle internalization, endosomal escape, and ATP-binding cassette (ABC) transporters inhibition. This integrated strategy overcomes PTX resistance, depletes CAF, and reduces stromal proteins, doubling the tumor inhibition rate compared to PL L -siGli1/PTX (88 % vs. 44 %, P < 0.0001). As the first exploration of branched ligand-functionalized PEGylation, this study demonstrates that BLIP platform (termed APL B ) offers a versatile, low-immunogenicity strategy for synergistic RNAi and chemotherapy, emerging as a promising “blip” solution to overcome the PEGylation-related immune challenges and chemoresistance in NSCLC. • BLIP , the first branched ligand-functionalized PEGylation avoids ABC phenomenon. • BLIP integrates dual targeting of NSCLC/CAF and RNAi-chemotherapy synergy . • BLIP inhibits ABC transporters, enhances cytotoxicity, depletes CAF stroma. • BLIP boosts tumor inhibition rate by 88 % , overcoming PTX resistance in NSCLC.