- Li, Yan-Ruide;
- Fang, Ying;
- Niu, Siyue;
- Zhu, Yichen;
- Chen, Yuning;
- Lyu, Zibai;
- Zhu, Enbo;
- Tian, Yanxin;
- Huang, Jie;
- Rezek, Valerie;
- Kitchen, Scott;
- Hsiai, Tzung;
- Zhou, Jin;
- Wang, Pin;
- Chai-Ho, Wanxing;
- Park, Sunmin;
- Seet, Christopher;
- Oliai, Caspian;
- Yang, Lili
Chimeric antigen receptor (CAR)-engineered T cell therapy holds promise for treating myeloid malignancies, but challenges remain in bone marrow (BM) infiltration and targeting BM-resident malignant cells. Current autologous CAR-T therapies also face manufacturing and patient selection issues, underscoring the need for off-the-shelf products. In this study, we characterize primary patient samples and identify a unique therapeutic opportunity for CAR-engineered invariant natural killer T (CAR-NKT) cells. Using stem cell gene engineering and a clinically guided culture method, we generate allogeneic CD33-directed CAR-NKT cells with high yield, purity, and robustness. In preclinical mouse models, CAR-NKT cells exhibit strong BM homing and effectively target BM-resident malignant blast cells, including CD33-low/negative leukemia stem and progenitor cells. Furthermore, CAR-NKT cells synergize with hypomethylating agents, enhancing tumor-killing efficacy. These cells also show minimal off-tumor toxicity, reduced graft-versus-host disease and cytokine release syndrome risks, and resistance to allorejection, highlighting their substantial therapeutic potential for treating myeloid malignancies.