Selective HLA knockdown and PD-L1 expression prevent allogeneic CAR-NK cell rejection and enhance safety and anti-tumor responses in xenograft mice
Allogeneic cellular immunotherapy exhibits promising efficacy for cancer treatment, but donor cell rejection remains a major barrier. Here, we systematically evaluate human leukocyte antigens (HLA) and immune checkpoints PD-L1, HLA-E, and CD47 in the rejection of allogeneic NK cells and identify CD8<sup>+</sup> T cells as the dominant cell type mediating allorejection. We demonstrate that a single gene construct that combines an shRNA that selectively interferes with HLA class I but not HLA-E expression, a chimeric antigen receptor (CAR), and PD-L1 or single-chain HLA-E (SCE) enables the one-step construction of allogeneic CAR-NK cells that evade host-mediated rejection both in vitro and in a xenograft mouse model. Furthermore, CAR-NK cells overexpressing PD-L1 or SCE effectively kill tumor cells through the upregulation of cytotoxic genes and reduced exhaustion and exhibit a favorable safety profile due to the decreased production of inflammatory cytokines involved in cytokine release syndrome. Thus, our approach represents a promising strategy in enabling "off-the-shelf" allogeneic cellular immunotherapies.
Authors
- Jerome Ritz (ORCID: https://orcid.org/0000-0001-5526-4669)
- Yuhao Qiang
- Minh Nguyen (ORCID: https://orcid.org/0009-0004-8020-9767)
- Khanhlinh Dinh (ORCID: https://orcid.org/0009-0007-6108-4147)
- Alaa Kassim Ali (ORCID: https://orcid.org/0000-0002-7490-0592)
- Xingyu Deng
- Jianzhu Chen (ORCID: https://orcid.org/0000-0002-5687-6154)
- Toni K. Choueiri (ORCID: https://orcid.org/0000-0002-9201-3217)
- Fuguo Liu (ORCID: https://orcid.org/0000-0002-1645-0976)
- Rizwan Romee (ORCID: https://orcid.org/0000-0001-9077-4736)
- Kewen Lei (ORCID: https://orcid.org/0000-0002-3586-682X)
- Yiming J. Zhang (ORCID: https://orcid.org/0000-0002-8102-3815)
- Mubin Tarannum (ORCID: https://orcid.org/0000-0002-4440-1565)
- James Dongjoo Ham (ORCID: https://orcid.org/0000-0003-0971-7940)
- Shaobo Yang (ORCID: https://orcid.org/0009-0006-5257-6775)
- Yingjie Zhao (ORCID: https://orcid.org/0000-0001-7170-2403)
Institutions
- Dana-Farber Cancer Institute (US)
- Dana-Farber/Harvard Cancer Center (US)
- Massachusetts Institute of Technology (US)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-10-08
- DOI
- https://doi.org/10.1038/s41467-025-63863-8
- Citations
- 8
- Primary Topic
- Immune Cell Function and Interaction
- Type
- article
- Field-Weighted Citation Impact
- 6.38
Funders
- Leukemia and Lymphoma Society
- Barr Foundation
- Parker Institute for Cancer Immunotherapy
- Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology
- National Institutes of Health
- National Cancer Institute