Targeting and cytotoxicity of chimeric antigen receptor T cells grafted with PD1 extramembrane domain

Experimental Hematology & Oncology

Research

Hu Chen¹  Lian Duan¹  Ang Zhang²  Shenyu Wang³  Lintao Shi⁴  Yikun Zhang⁴  Lei Liu⁴  Mingyi He⁴  Quanjun Wang⁵  Jie Du⁵  Lei Xu⁶  Yao Sun⁶  Liangding Hu⁶  Bin Zhang⁶  Yijian Zhang⁶  Yangyang Lei⁶  Yang Yang⁶  Long Zhao⁶ 

[1] Academy of Military Medical Sciences, Academy of Military Sciences, 100850, Beijing, PR China;Academy of Military Medical Sciences, Academy of Military Sciences, 100850, Beijing, PR China;Department of Hematology, Strategic Support Force Medical Center, Beijing, China;Academy of Military Medical Sciences, Academy of Military Sciences, 100850, Beijing, PR China;Senior Department of Hematology, the Fifth Medical Center of Chinese PLA General Hospital, 100071, Beijing, PR China;Department of Hematology, Strategic Support Force Medical Center, Beijing, China;SAFE Pharmaceutical Research Institute Co., Ltd, Beijing, China;Senior Department of Hematology, the Fifth Medical Center of Chinese PLA General Hospital, 100071, Beijing, PR China;

关键词: Chimeric antigen receptor T cell;    PD1;    Off-target toxicity;    Hinge domain;    Tumor microenvironment;   

DOI  :  10.1186/s40164-023-00438-7

 received in 2023-04-02, accepted in 2023-08-22,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundImmunosuppression induced by programmed cell death protein 1 (PD1) presents a significant constraint on the effectiveness of chimeric antigen receptor (CAR)-T therapy. The potential of combining PD1/PDL1 (Programmed cell death 1 ligand 1) axis blockade with CAR-T cell therapy is promising. However, developing a highly efficient and minimally toxic approach requires further exploration. Our attempt to devise a novel CAR structure capable of recognizing both tumor antigens and PDL1 encountered challenges since direct targeting of PDL1 resulted in systemic adverse effects.MethodsIn this research, we innovatively engineered novel CARs by grafting the PD1 domain into a conventional second-generation (2G) CAR specifically targeting CD19. These CARs exist in two distinct forms: one with PD1 extramembrane domain (EMD) directly linked to a transmembrane domain (TMD), referred to as PE CAR, and the other with PD1 EMD connected to a TMD via a CD8 hinge domain (HD), known as PE8HT CAR. To evaluate their efficacy, we conducted comprehensive assessments of their cytotoxicity, cytokine release, and potential off-target effects both in vitro and in vivo using tumor models that overexpress CD19/PDL1.ResultsThe findings of our study indicate that PE CAR demonstrates enhanced cytotoxicity and reduced cytokine release specifically towards CD19 + PDL1 + tumor cells, without off-target effects to CD19-PDL1 + tumor cells, in contrast to 2G CAR-T cells. Additionally, PE CAR showed ameliorative differentiation, exhaustion, and apoptosis phenotypes as assessed by flow cytometry, RNA-sequencing, and metabolic parameter analysis, after encountering CD19 + PDL1 + tumor cells.ConclusionOur results revealed that CAR grafted with PD1 exhibits enhanced antitumor activity with lower cytokine release and no PD1-related off-target toxicity in tumor models that overexpress CD19 and PDL1. These findings suggest that our CAR design holds the potential for effectively addressing the PD1 signal.

【 授权许可】

CC BY   
© YUMED Inc. and BioMed Central Ltd. 2023

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