Engineering allorejection-resistant CAR-NKT cells from hematopoietic stem cells for off-the-shelf cancer immunotherapy

doi:10.1016/j.ymthe.2024.04.005

. 2024 Jun 5;32(6):1849-1874.

doi: 10.1016/j.ymthe.2024.04.005. Epub 2024 Apr 6.

Engineering allorejection-resistant CAR-NKT cells from hematopoietic stem cells for off-the-shelf cancer immunotherapy

Yan-Ruide Li¹, Yang Zhou¹, Jiaji Yu¹, Yichen Zhu¹, Derek Lee¹, Enbo Zhu², Zhe Li¹, Yu Jeong Kim¹, Kuangyi Zhou¹, Ying Fang¹, Zibai Lyu¹, Yuning Chen¹, Yanxin Tian¹, Jie Huang¹, Xinjian Cen¹, Tiffany Husman¹, Jae Min Cho³, Tzung Hsiai³, Jin J Zhou⁴, Pin Wang⁵, Benjamin R Puliafito⁶, Sarah M Larson⁷, Lili Yang⁸

Affiliations

¹ Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA; Division of Cardiology, Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ Division of Cardiology, Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁴ Department of Biostatistics, Fielding School of Public Health, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁵ Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, CA 90089, USA.
⁶ Department of Hematology and Oncology, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁷ Department of Internal Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Centre, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁸ Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Centre, University of California, Los Angeles, Los Angeles, CA 90095, USA; Eli and Edythe Broad Centre of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA 90095, USA; Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: liliyang@ucla.edu.

PMID: 38584391
PMCID: PMC11184334 (available on 2025-06-05)
DOI: 10.1016/j.ymthe.2024.04.005

Engineering allorejection-resistant CAR-NKT cells from hematopoietic stem cells for off-the-shelf cancer immunotherapy

Yan-Ruide Li et al. Mol Ther. 2024.

. 2024 Jun 5;32(6):1849-1874.

doi: 10.1016/j.ymthe.2024.04.005. Epub 2024 Apr 6.

Authors

Affiliations

¹ Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA; Division of Cardiology, Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ Division of Cardiology, Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁴ Department of Biostatistics, Fielding School of Public Health, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁵ Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, CA 90089, USA.
⁶ Department of Hematology and Oncology, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁷ Department of Internal Medicine, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Centre, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁸ Department of Microbiology, Immunology & Molecular Genetics, University of California, Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Centre, University of California, Los Angeles, Los Angeles, CA 90095, USA; Eli and Edythe Broad Centre of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA 90095, USA; Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: liliyang@ucla.edu.

PMID: 38584391
PMCID: PMC11184334 (available on 2025-06-05)
DOI: 10.1016/j.ymthe.2024.04.005

Abstract

The clinical potential of current FDA-approved chimeric antigen receptor (CAR)-engineered T (CAR-T) cell therapy is encumbered by its autologous nature, which presents notable challenges related to manufacturing complexities, heightened costs, and limitations in patient selection. Therefore, there is a growing demand for off-the-shelf universal cell therapies. In this study, we have generated universal CAR-engineered NKT (^UCAR-NKT) cells by integrating iNKT TCR engineering and HLA gene editing on hematopoietic stem cells (HSCs), along with an ex vivo, feeder-free HSC differentiation culture. The ^UCAR-NKT cells are produced with high yield, purity, and robustness, and they display a stable HLA-ablated phenotype that enables resistance to host cell-mediated allorejection. These ^UCAR-NKT cells exhibit potent antitumor efficacy to blood cancers and solid tumors, both in vitro and in vivo, employing a multifaceted array of tumor-targeting mechanisms. These cells are further capable of altering the tumor microenvironment by selectively depleting immunosuppressive tumor-associated macrophages and myeloid-derived suppressor cells. In addition, ^UCAR-NKT cells demonstrate a favorable safety profile with low risks of graft-versus-host disease and cytokine release syndrome. Collectively, these preclinical studies underscore the feasibility and significant therapeutic potential of ^UCAR-NKT cell products and lay a foundation for their translational and clinical development.

Keywords: CRISPR-Cas9 gene editing; T cell receptor gene engineering; allogeneic cell therapy; allorejection resistance; chimeric antigen receptor engineering; hematopoietic stem cell engineering; invariant natural killer T cell; off-the-shelf cancer immunotherapy; tumor microenvironment; universal CAR-engineered NKT cell.

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Conflict of interest statement

Declaration of interests Y.-R.L., Y. Zhou, J.Y., D.L., Z.L., Y.J.K., P.W., and L.Y. are inventors on patents relating to this study filed by UCLA. Y. Zhou is currently an employee of Amberstone Biosciences. J.Y. is currently an employee of Appia Bio. Z.L. is currently an employee of Allogene. Y.J.K. is currently an employee of Nkarta. P.W. is a co-founder, stockholder, and advisory board member of Simnova Bio, TCRCure Biopharma, Appia Bio, and is a scientific advisor to Grit Biotechnolgoy. L.Y. is a scientific advisor to AlzChem and Amberstone Biosciences, and a co-founder, stockholder, and advisory board member of Appia Bio. Appia Bio licensed some patents relating to this study from UCLA. None of the declared companies contributed to or directed any of the research reported in this article. The remaining authors declare no competing interests.

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References

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1. Finck A.V., Blanchard T., Roselle C.P., Golinelli G., June C.H. Engineered cellular immunotherapies in cancer and beyond. Nat. Med. 2022;28:678–689. doi: 10.1038/s41591-022-01765-8. - DOI - PMC - PubMed
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[1] June C.H., O’Connor R.S., Kawalekar O.U., Ghassemi S., Milone M.C. CAR T cell immunotherapy for human cancer. Science. 2018;359:1361–1365. doi: 10.1126/science.aar6711. - DOI - PubMed

[2] June C.H., O’Connor R.S., Kawalekar O.U., Ghassemi S., Milone M.C. CAR T cell immunotherapy for human cancer. Science. 2018;359:1361–1365. doi: 10.1126/science.aar6711. - DOI - PubMed

[3] Sterner R.C., Sterner R.M. CAR-T cell therapy: current limitations and potential strategies. Blood Cancer J. 2021;11:69. doi: 10.1038/s41408-021-00459-7. - DOI - PMC - PubMed

[4] Sterner R.C., Sterner R.M. CAR-T cell therapy: current limitations and potential strategies. Blood Cancer J. 2021;11:69. doi: 10.1038/s41408-021-00459-7. - DOI - PMC - PubMed

[5] Labanieh L., Mackall C.L. CAR immune cells: design principles, resistance and the next generation. Nature. 2023;614:635–648. doi: 10.1038/s41586-023-05707-3. - DOI - PubMed

[6] Labanieh L., Mackall C.L. CAR immune cells: design principles, resistance and the next generation. Nature. 2023;614:635–648. doi: 10.1038/s41586-023-05707-3. - DOI - PubMed

[7] Finck A.V., Blanchard T., Roselle C.P., Golinelli G., June C.H. Engineered cellular immunotherapies in cancer and beyond. Nat. Med. 2022;28:678–689. doi: 10.1038/s41591-022-01765-8. - DOI - PMC - PubMed

[8] Finck A.V., Blanchard T., Roselle C.P., Golinelli G., June C.H. Engineered cellular immunotherapies in cancer and beyond. Nat. Med. 2022;28:678–689. doi: 10.1038/s41591-022-01765-8. - DOI - PMC - PubMed

[9] Li Y.-R., Dunn Z.S., Yu Y., Li M., Wang P., Yang L. Advancing cell-based cancer immunotherapy through stem cell engineering. Cell Stem Cell. 2023;30:592–610. doi: 10.1016/j.stem.2023.02.009. - DOI - PMC - PubMed

[10] Li Y.-R., Dunn Z.S., Yu Y., Li M., Wang P., Yang L. Advancing cell-based cancer immunotherapy through stem cell engineering. Cell Stem Cell. 2023;30:592–610. doi: 10.1016/j.stem.2023.02.009. - DOI - PMC - PubMed

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Engineering allorejection-resistant CAR-NKT cells from hematopoietic stem cells for off-the-shelf cancer immunotherapy

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Engineering allorejection-resistant CAR-NKT cells from hematopoietic stem cells for off-the-shelf cancer immunotherapy

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