Yun Chang, Kunming Shao, Huiyang Li, Gyuhyung Jin, R. Timothy Bentley, Robyn McCain, Christa J. Crain, Jingqiao Shen, Yan Tan, Po-Yu Liang, Haley A. Harper, Sandra Torregrosa-Allen, Bennett D. Elzey, Isabelle F. Vanhaezebrouck, Aaron A. Cohen-Gadol, Cheng Dong, Yu Zhu, Yuewei Wang, Jie Luo, Xiaojun Lian, Xiaoping Bao
Despite their notable abundance in circulation and biological significance in the tumour microenvironment, direct genetic programming of neutrophils has been challenging. Here we develop a neutrophil-specific modified RNA translation platform, termed NeuSMRT, that enables the expression of chimeric antigen receptors (CARs) in primary neutrophils. NeuSMRT combines engineered extracellular vesicles or lipid nanoparticles for modified RNA delivery with a microRNA-responsive L7Ae:k-turn switch to restrict protein translation to neutrophils. In a syngeneic glioma model, CAR-neutrophils produced in vivo significantly inhibit tumour growth and prolong survival, accompanied by enhanced T cell recruitment and activation, and reduced immunosuppression of myeloid cells in the tumour microenvironment. CAR-neutrophils further enhance the efficacy of chemotherapy and CAR-T therapy. Furthermore, CAR-neutrophils display antitumour activities in a humanized glioblastoma mouse model. The feasibility and safety of NeuSMRT are also demonstrated in experimental dogs. These findings establish a programmable neutrophil platform for cancer immunotherapy.