Synergistic innate-adaptive immunity by NKG2D-specific CAR-macrophages drives durable remission in hepatocellular carcinoma

作者：Zihao Zhao, Wenjing Zheng, Yang He, Han Zhang, Lingling Zhang, Yi Huo, Junwei Jiang, Chen Zhang, Haohan Lyu, Weiwei Qin, Chen Liu, Feng Chang, Lequn Shan, Tao Wang, Wenjie Song · 发表于：Molecular Cancer · 年份：2025 · DOI：10.1186/s12943-025-02538-w · 被引用次数：5 · 研究领域：Immune Cell Function and Interaction、Immune cells in cancer、CAR-T cell therapy research

BACKGROUND: Hepatocellular carcinoma (HCC) immunotherapy is limited by antigenic heterogeneity and an immunosuppressive microenvironment. This study engineered chimeric antigen receptor macrophages (CAR-Ms) targeting stress-inducible NKG2D ligands (NKG2DLs), broadly overexpressed in HCC, to enhance phagocytic clearance and remodel immunity. METHODS: NKG2DL expression in HCC and association with survival were analyzed. CAR-Ms were constructed by fusing the NKG2D extracellular domain to FcγRI signaling. In vitro assays assessed phagocytosis, cytokine secretion, signaling, and T cell interactions. Therapeutic efficacy was evaluated in immunocompetent mice bearing subcutaneous, orthotopic, or metastatic HCC models, with or without anti-PD-L1. Tumor progression, immunity, and survival were analyzed via bioluminescence imaging, flow cytometry, histopathology, and serum biochemistry. Statistics analyses were performed using t-tests, ANOVA, and log-rank tests. RESULTS: NKG2DLs were significantly upregulated in human HCC and correlated with poor prognosis. CAR-Ms selectively engulfed NKG2DL⁺ tumor cells, polarized to an M1 phenotype, and activated PI3K-AKT and cGAS-STING pathways, driving phagocytosis and pro-inflammatory cytokines secretion. They enhanced T cell chemokines (Cxcl10, Ccl5) and antigen presentation, boosting T cell recruitment and activation in vitro. In subcutaneous models, CAR-Ms suppressed tumor growth, reprogrammed tumor-associated myeloid cells toward M1, and induc...