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

Yan-Ruide Li; Yang Zhou; Jiaji Yu; Yichen Zhu; Derek Lee; Enbo Zhu; Zhe Li; Yu Jeong Kim; Kuangyi Zhou; Ying Fang; Zibai Lyu; Yuning Chen; Yanxin Tian; Jie Huang; Xinjian Cen; Tiffany Husman; Jae Min Cho; Tzung Hsiai; Jin J Zhou; Pin Wang; Benjamin R Puliafito; Sarah M Larson; Lili Yang

doi:10.1016/j.ymthe.2024.04.005

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

Mol Ther. 2024 Apr 6:S1525-0016(24)00221-1. doi: 10.1016/j.ymthe.2024.04.005. Online ahead of print.

Authors

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
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.