Frontiers in Immunology (Nov 2024)

Hypoimmunogenic CD19 CAR-NK cells derived from embryonic stem cells suppress the progression of human B-cell malignancies in xenograft animals

  • Qi Zhang,
  • Chengxiang Xia,
  • Qitong Weng,
  • Leqiang Zhang,
  • Yao Wang,
  • Yanhong Liu,
  • Xiujuan Zheng,
  • Yunqing Lin,
  • Yi Chen,
  • Yiyuan Shen,
  • Hanmeng Qi,
  • Lijuan Liu,
  • Yanping Zhu,
  • Min Zhang,
  • Dehao Huang,
  • Fangxiao Hu,
  • Mengyun Zhang,
  • Mengyun Zhang,
  • Hui Zeng,
  • Jinyong Wang,
  • Jinyong Wang,
  • Tongjie Wang,
  • Tongjie Wang

DOI
https://doi.org/10.3389/fimmu.2024.1504459
Journal volume & issue
Vol. 15

Abstract

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BackgroundChimeric antigen receptor (CAR) engineered natural killer (NK) cells exhibit advantages such as MHC-independent recognition and strong anti-tumor functions. However, allogeneic CAR-NK cells derived from human tissues are heterogeneous and susceptible to clearance by hosts.MethodsWe generated a B2M knockout, HLA-E and CD19 CAR ectopic expressing embryonic stem cell (ESC) line, which differentiated normally and gave rise to homogeneous CD19 CAR-NK (CD19 CAR-UiNK) cells using an organoid aggregate induction method. The CD19 CAR-UiNK were co-cultured with T cells or NK cells derived from peripheral blood mononuclear cells (PBMC) with the mismatched HLA to evaluate the immunogenicity of CD19 CAR-UiNK cells. We further assessed the therapeutic effects of CD19 CAR-UiNK cells on CD19+ tumor cells through in vitro cytotoxicity assays and in vivo animal models.ResultsThe CD19 CAR-UiNK cells exhibited typical expression patterns of activating and inhibitory receptors, and crucial effector molecules of NK cells, similar to those of unmodified NK cells. In co-culture assays, the CD19 CAR-UiNK cells evaded allogeneic T cell response and suppressed allogeneic NK cell response. Functionally, the CD19 CAR-UiNK cells robustly secreted IFN-γ and TNF-α, and upregulated CD107a upon stimulation with Nalm-6 tumor cells. The CD19 CAR-UiNK cells effectively eliminated CD19+ tumor cells in vitro, including B-cell cancer cell lines and primary tumor cells from human B-cell leukemia and lymphoma. Further, the CD19 CAR-UiNK cells exhibited strong anti-tumor activity in xenograft animals.ConclusionWe offer a strategy for deriving homogeneous and hypoimmunogenic CD19 CAR-iNK cells with robust anti-tumor effects from ESCs. Our study has significant implications for developing hypoimmunogenic CD19 CAR-NK cell therapy using human ESC as an unlimited cell source.

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