Generation of allogeneic CAR-NKT cells from hematopoietic stem and progenitor cells using a clinically guided culture method

Yan-Ruide Li; Yang Zhou; Jiaji Yu; Yu Jeong Kim; Miao Li; Derek Lee; Kuangyi Zhou; Yuning Chen; Yichen Zhu; Yu-Chen Wang; Zhe Li; Yanqi Yu; Zachary Spencer Dunn; Wenbin Guo; Xinjian Cen; Tiffany Husman; Aarushi Bajpai; Adam Kramer; Matthew Wilson; Ying Fang; Jie Huang; Shuo Li; Yonggang Zhou; Yuchong Zhang; Zoe Hahn; Enbo Zhu; Feiyang Ma; Calvin Pan; Aldons J Lusis; Jin J Zhou; Christopher S Seet; Donald B Kohn; Pin Wang; Xianghong Jasmine Zhou; Matteo Pellegrini; Benjamin R Puliafito; Sarah M Larson; Lili Yang

doi:10.1038/s41587-024-02226-y

Generation of allogeneic CAR-NKT cells from hematopoietic stem and progenitor cells using a clinically guided culture method

Nat Biotechnol. 2024 May 14. doi: 10.1038/s41587-024-02226-y. Online ahead of print.

Authors

Yan-Ruide Li^#¹, Yang Zhou^#¹, Jiaji Yu^#¹, Yu Jeong Kim¹, Miao Li¹, Derek Lee¹, Kuangyi Zhou¹, Yuning Chen¹, Yichen Zhu¹, Yu-Chen Wang², Zhe Li¹, Yanqi Yu¹, Zachary Spencer Dunn³, Wenbin Guo⁴, Xinjian Cen¹, Tiffany Husman¹, Aarushi Bajpai¹, Adam Kramer¹, Matthew Wilson¹, Ying Fang¹, Jie Huang¹, Shuo Li⁵, Yonggang Zhou⁵, Yuchong Zhang¹, Zoe Hahn¹, Enbo Zhu¹, Feiyang Ma⁶, Calvin Pan², Aldons J Lusis^{1

2

7}, Jin J Zhou⁸, Christopher S Seet^{9

10

11

12}, Donald B Kohn^{1

9

13}, Pin Wang³, Xianghong Jasmine Zhou⁵, Matteo Pellegrini^{4

14

15}, Benjamin R Puliafito¹⁶, Sarah M Larson^{10

17}, Lili Yang^{18

19

20

21}

Affiliations

¹ Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA, USA.
² Department of Medicine, Division of Cardiology, University of California, Los Angeles, Los Angeles, CA, USA.
³ Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, CA, USA.
⁴ Bioinformatics Interdepartmental Program, University of California, Los Angeles, Los Angeles, CA, USA.
⁵ Department of Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
⁶ Department of Cell and Developmental Biology, Feinberg School of Medicine, Northwestern University, Chicago, IL, USA.
⁷ Human Genetics, University of California, Los Angeles, Los Angeles, CA, USA.
⁸ Department of Biostatistics, Fielding School of Public Health, University of California, Los Angeles, Los Angeles, CA, USA.
⁹ Eli and Edythe Broad Centre of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA, USA.
¹⁰ Jonsson Comprehensive Cancer Centre, University of California, Los Angeles, Los Angeles, CA, USA.
¹¹ Department of Medicine, Division of Hematology/Oncology, University of California, Los Angeles, Los Angeles, CA, USA.
¹² Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, USA.
¹³ Department of Pediatrics, Division of Hematology/Oncology, University of California, Los Angeles, Los Angeles, CA, USA.
¹⁴ Institute for Quantitative and Computational Biosciences-The Collaboratory, University of California, Los Angeles, Los Angeles, CA, USA.
¹⁵ Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, Los Angeles, CA, USA.
¹⁶ Department of Hematology and Oncology, University of California, Los Angeles, Los Angeles, CA, USA.
¹⁷ Department of Internal Medicine, University of California, Los Angeles, Los Angeles, CA, USA.
¹⁸ Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, Los Angeles, CA, USA. [email protected].
¹⁹ Eli and Edythe Broad Centre of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA, USA. [email protected].
²⁰ Jonsson Comprehensive Cancer Centre, University of California, Los Angeles, Los Angeles, CA, USA. [email protected].
²¹ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, USA. [email protected].

^# Contributed equally.

PMID: 38744947
DOI: 10.1038/s41587-024-02226-y

Abstract

Cancer immunotherapy with autologous chimeric antigen receptor (CAR) T cells faces challenges in manufacturing and patient selection that could be avoided by using 'off-the-shelf' products, such as allogeneic CAR natural killer T (^AlloCAR-NKT) cells. Previously, we reported a system for differentiating human hematopoietic stem and progenitor cells into ^AlloCAR-NKT cells, but the use of three-dimensional culture and xenogeneic feeders precluded its clinical application. Here we describe a clinically guided method to differentiate and expand IL-15-enhanced ^AlloCAR-NKT cells with high yield and purity. We generated ^AlloCAR-NKT cells targeting seven cancers and, in a multiple myeloma model, demonstrated their antitumor efficacy, expansion and persistence. The cells also selectively depleted immunosuppressive cells in the tumor microenviroment and antagonized tumor immune evasion via triple targeting of CAR, TCR and NK receptors. They exhibited a stable hypoimmunogenic phenotype associated with epigenetic and signaling regulation and did not induce detectable graft versus host disease or cytokine release syndrome. These properties of ^AlloCAR-NKT cells support their potential for clinical translation.

Abstract

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