Allogeneic CAR-T cells with of HLA-A/B and TRAC disruption exhibit promising antitumor capacity against B cell malignancies

Xinfeng Chen; Binghe Tan; Haizhou Xing; Xuan Zhao; Yu Ping; Zhen Zhang; Jianmin Huang; Xiujuan Shi; Na Zhang; Boxu Lin; Weijie Cao; Xin Li; Xudong Zhang; Ling Li; Zhongxing Jiang; Mingzhi Zhang; Wei Li; Mingyao Liu; Bing Du; Yi Zhang

doi:10.1007/s00262-023-03586-1

Allogeneic CAR-T cells with of HLA-A/B and TRAC disruption exhibit promising antitumor capacity against B cell malignancies

Cancer Immunol Immunother. 2024 Jan 17;73(1):13. doi: 10.1007/s00262-023-03586-1.

Authors

Xinfeng Chen^#^{1

2}, Binghe Tan^#^{3

4}, Haizhou Xing^#⁵, Xuan Zhao^{1

2}, Yu Ping¹, Zhen Zhang¹, Jianmin Huang¹, Xiujuan Shi⁴, Na Zhang³, Boxu Lin³, Weijie Cao⁵, Xin Li², Xudong Zhang², Ling Li², Zhongxing Jiang⁵, Mingzhi Zhang², Wei Li⁴, Mingyao Liu³, Bing Du⁶, Yi Zhang^{7

8

9

10

11}

Affiliations

¹ Biotherapy Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China.
² Cancer Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China.
³ Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, 200241, China.
⁴ BRL Medicine Inc, Shanghai, 201109, China.
⁵ Department of Hematology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China.
⁶ Shanghai Frontiers Science Center of Genome Editing and Cell Therapy, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, 200241, China. bdu@bio.ecnu.edu.cn.
⁷ Biotherapy Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China. yizhang@zzu.edu.cn.
⁸ Cancer Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China. yizhang@zzu.edu.cn.
⁹ State Key Laboratory of Esophageal Cancer Prevention & Treatment, Zhengzhou, 450052, Henan, China. yizhang@zzu.edu.cn.
¹⁰ School of Life Sciences, Zhengzhou University, Zhengzhou, 450052, Henan, China. yizhang@zzu.edu.cn.
¹¹ Engineering Key Laboratory for Cell Therapy of Henan Province, Zhengzhou, 450052, Henan, China. yizhang@zzu.edu.cn.

^# Contributed equally.

Abstract

Background: Although chimeric antigen receptor T (CAR-T) cells have been proven to be an effective way of treating B cell malignancies, a lot of patients could not benefit from it because of failure in CAR-T cell manufacturing, disease progression, and unaffordable price. The study aimed to explore universal CAR-T cell products to extend the clinical accessibility.

Methods: The antitumor activity of CRISPR/Cas9-edited allogeneic anti-CD19 CAR-T (CAR-T19) cells was assessed in vitro, in animal models, and in patients with relapsed/refractory (R/R) acute B cell lymphoblastic leukemia (B-ALL) or diffuse large B cell lymphoma.

Results: B2M^-/TRAC^- universal CAR-T19 (U-CAR-T19) cells exhibited powerful anti-leukemia abilities both in vitro and in animal models, as did primary CD19⁺ leukemia cells from leukemia patients. However, expansion, antitumor efficacy, or graft-versus-host-disease (GvHD) was not observed in six patients with R/R B cell malignancies after U-CAR-T19 cell infusion. Accordingly, significant activation of natural killer (NK) cells by U-CAR-T19 cells was proven both clinically and in vitro. HLA-A^-/B^-/TRAC^- novel CAR-T19 (nU-CAR-T19) cells were constructed with similar tumoricidal capacity but resistance to NK cells in vitro. Surprisingly, robust expansion of nU-CAR-T19 cells, along with rapid eradication of CD19⁺ abnormal B cells, was observed in the peripheral blood and bone marrow of another three patients with R/R B-ALL. The patients achieved complete remission with no detectable minimal residual disease 14 days after the infusion of nU-CAR-T19 cells. Two of the three patients had grade 2 cytokine release syndrome, which were managed using an IL-6 receptor blocker. Most importantly, GvHD was not observed in any patient, suggesting the safety of TRAC-disrupted CAR-T cells generated using the CRISPR/Cas9 method for clinical application.

Conclusions: The nU-CAR-T19 cells showed a strong response in R/R B-ALL. nU-CAR-T19 cells have the potential to be a promising new approach for treating R/R B cell malignancies.

Keywords: B cell malignancies; CRISPR/Cas9; HLA; NK cell; Universal CAR-T cell.

MeSH terms

Animals
Antibodies
Antigens, CD19
Graft vs Host Disease*
HLA-A Antigens
Hematopoietic Stem Cell Transplantation*
Humans
Leukemia*
Leukemia, Lymphocytic, Chronic, B-Cell*
Receptors, Chimeric Antigen* / genetics
T-Lymphocytes

Substances

Receptors, Chimeric Antigen
Antibodies
Antigens, CD19
HLA-A Antigens

Abstract

MeSH terms

Substances

Grants and funding