CAR-T cells dual-target CD123 and NKG2DLs to eradicate AML cells and selectively target immunosuppressive cells

Xin Jin; Danni Xie; Rui Sun; Wenyi Lu; Xia Xiao; Yibing Yu; Juanxia Meng; Mingfeng Zhao

doi:10.1080/2162402X.2023.2248826

CAR-T cells dual-target CD123 and NKG2DLs to eradicate AML cells and selectively target immunosuppressive cells

Oncoimmunology. 2023 Aug 26;12(1):2248826. doi: 10.1080/2162402X.2023.2248826. eCollection 2023.

Authors

Xin Jin^{1

2

3}, Danni Xie⁴, Rui Sun¹, Wenyi Lu³, Xia Xiao³, Yibing Yu⁴, Juanxia Meng³, Mingfeng Zhao³

Affiliations

¹ School of Medicine, Nankai University, Tianjin, China.
² Liangzhu Laboratory, Zhejiang University Medical Center, Hangzhou, China.
³ Department of Hematology, Tianjin First Central Hospital, School of Medicine, Nankai University, Tianjin, China.
⁴ First Central Clinical College, Tianjin Medical University, Tianjin, China.

Abstract

Chimeric antigen receptor (CAR)-T cells have not made significant progress in the treatment of acute myeloid leukemia (AML) in earlyclinical studies. This lack of progress could be attributed in part to the immunosuppressive microenvironment of AML, such as monocyte-like myeloid-derived suppressor cells (M-MDSCs) and alternatively activated macrophages (M2 cells), which can inhibit the antitumor activity of CAR-T cells. Furthermore, AML cells are usually heterogeneous, and single-target CAR-T cells may not be able to eliminate all AML cells, leading to disease relapse. CD123 and NKG2D ligands (NKG2DLs) are commonly used targets for CAR-T therapy of AML, and M-MDSCs and M2 cells express both antigens. We developed dual-targeted CAR-T (123NL CAR-T) cells targeting CD123 and NKG2DL by various structural optimization screens. Our study reveals that 123NL CAR-T cells eradicate AML cells and selectively target immunosuppressive cells. A highly compact marker/suicide gene, RQR8, which binds targeting epitopes of CD34 and CD20 antigens, was also incorporated in front of the CAR structure. The binding of Rituximab to RQR8 leads to the elimination of 123NL CAR-T cells and cessation of their cytotoxicity. In conclusion, we successfully developed dual effects of 123NL CAR-T cells against tumor cells and immunosuppressive cells, which can avoid target escape and resist the effects of immunosuppressive microenvironment.

Keywords: AML; CAR-T; CD123; NKG2DL; immunotherapy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Humans
Interleukin-3 Receptor alpha Subunit
Leukemia, Myeloid, Acute* / therapy
Ligands
Receptors, Chimeric Antigen* / genetics
T-Lymphocytes
Tumor Microenvironment

Substances

Interleukin-3 Receptor alpha Subunit
Ligands
Receptors, Chimeric Antigen

Grants and funding

This work was supported by grants from the General Project of the National Natural Science Foundation of China (81970180 to MZ), the Science and Technology Project of Tianjin Municipal Health Committee (TJWJ2022QN030 to MZ), Key projects of Tianjin Applied Basic Research and Multi-Investment Fund (21JCZDJC01240), Science and Technology Project of Tianjin Municipal Health Committee (TJWJ2022XK018 to MZ), the Key Science and Technology Support Project of Tianjin Science and Technology Bureau (20YFZCSY00800 to MZ), Tianjin Health Bureau Project (ZC20074 to XX), and Tianjin Key Medical Discipline (Specialty) Construction Project (TJYXZDXK-056B), as well as China Postdoctoral Science Foundation (2022M722748).