Reprogramming Tumor-Associated Neutrophils to Enhance Radio-Immunotherapy

As central effector cells in innate immunity, neutrophils are rapidly recruited to tumors following radiotherapy (RT). However, their frequent polarization toward the pro-tumor N2 phenotype and the formation of metastasis-promoting neutrophil extracellular traps (NETs) significantly limit treatment outcomes. To reprogram these RT-recruited tumor-associated neutrophils (TANs), we develop injectable hydrogel microspheres (HMPs) co-loaded with a TLR4 agonist (lipopolysaccharide) and a PAD4 inhibitor (GSK484). The resulting L/G@HMPs effectively redirect TAN polarization toward the anti-tumor N1 phenotype, inhibit NET formation, and extend neutrophil lifespan beyond 72 h. In murine tumor models, the combination of RT and L/G@HMPs triggers robust innate and adaptive immune responses, marked by substantial accumulation of N1-polarized TANs and CD8⁺ T cells within tumors, leading to potent tumor eradication. This study presents a hydrogel-based strategy that concurrently modulates neutrophil lifespan, phenotype, and NETs, thereby transforming tumor-promoting neutrophils into anti-tumor allies for enhanced radio-immunotherapy.