CRISPR knock out of programmed cell death protein 1 enhances anti-tumor activity of cytotoxic T lymphocytes

Zhao, Zhihui; Shi, Long; Zhang, Wei; Han, Jinsheng; Zhang, Shaohui; Fu, Zexian; Cai, Jianhui

doi:10.18632/oncotarget.23730

Cited by 81 publications

(59 citation statements)

References 19 publications

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“…However, these therapies face multiple challenges, including tumor accessibility, tumor-dependent inhibitory signals, and a microenvironment that is hostile to T cells. To overcome these obstacles, CAR T cells have been engineered to remove inhibitory receptors [1][2][3] , to express costimulatory molecules 3 and chemokine receptors 4,5 , or to secrete immunostimulatory factors such as checkpoint inhibitors 6 or cytokines 7 . In particular, cytokine secretion by so called Armored or Truck CAR T cells 7,8 is a highly promising strategy for improving CAR T cell function.…”

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confidence: 99%

Repurposing endogenous immune pathways to tailor and control chimeric antigen receptor T cell functionality

et al. 2019

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Endowing chimeric antigen receptor (CAR) T cells with additional potent functionalities holds strong potential for improving their antitumor activity. However, because potency could be deleterious without control, these additional features need to be tightly regulated. Immune pathways offer a wide array of tightly regulated genes that can be repurposed to express potent functionalities in a highly controlled manner. Here, we explore this concept by repurposing TCR, CD25 and PD1, three major players of the T cell activation pathway. We insert the CAR into the TCRα gene (TRAC CAR), and IL-12P70 into either IL2Rα or PDCD1 genes. This process results in transient, antigen concentration-dependent IL-12P70 secretion, increases TRAC CAR T cell cytotoxicity and extends survival of tumor-bearing mice. This gene network repurposing strategy can be extended to other cellular pathways, thus paving the way for generating smart CAR T cells able to integrate biological inputs and to translate them into therapeutic outputs in a highly regulated manner.

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confidence: 99%

Repurposing endogenous immune pathways to tailor and control chimeric antigen receptor T cell functionality

et al. 2019

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“…Improved effector functions have also been reported following PDCD1 gene editing in virus-specific cytotoxic T lymphocytes (CTL) 23 24 and in myeloma-specific CTL. 25 In melanoma, the superior antitumor efficacy of PDCD1-edited human-specific CTL has not been formally documented so far. In this study, we demonstrated the feasibility of PDCD1 gene editing in high avidity effector T cells, specific for the Melan-A antigen, using electroporation of ribonucleic complexes.…”

Section: Introductionmentioning

confidence: 99%

Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytes

Marotte

Simon

Vignard

et al. 2020

J Immunother Cancer

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BackgroundGenome editing offers unique perspectives for optimizing the functional properties of T cells for adoptive cell transfer purposes. So far,PDCD1editing has been successfully tested mainly in chimeric antigen receptor T (CAR-T) cells and human primary T cells. Nonetheless, for patients with solid tumors, the adoptive transfer of effector memory T cells specific for tumor antigens remains a relevant option, and the use of high avidity T cells deficient for programmed cell death-1 (PD-1) expression is susceptible to improve the therapeutic benefit of these treatments.MethodsHere we used the transfection of CAS9/sgRNA ribonucleoproteic complexes to editPDCD1gene in human effector memory CD8+T cells specific for the melanoma antigen Melan-A. We cloned edited T cell populations and validatedPDCD1editing through sequencing and cytometry in each T cell clone, together with T-cell receptor (TCR) chain’s sequencing. We also performed whole transcriptomic analyses on wild-type (WT) and edited T cell clones. Finally, we documented in vitro and in vivo through adoptive transfer in NOD scid gamma (NSG) mice, the antitumor properties of WT and PD-1KO T cell clones, expressing the same TCR.ResultsHere we demonstrated the feasibility to editPDCD1gene in human effector memory melanoma-specific T lymphocytes. We showed that PD-1 expression was dramatically reduced or totally absent onPDCD1-edited T cell clones. Extensive characterization of a panel of T cell clones expressing the same TCR and exhibiting similar functional avidity demonstrated superior antitumor reactivity against a PD-L1 expressing melanoma cell line. Transcriptomic analysis revealed a downregulation of genes involved in proliferation and DNA replication in PD-1-deficient T cell clones, whereas genes involved in metabolism and cell signaling were upregulated. Finally, we documented the superior ability of PD-1-deficient T cells to significantly delay the growth of a PD-L1 expressing human melanoma tumor in an NSG mouse model.ConclusionThe use of such lymphocytes for adoptive cell transfer purposes, associated with other approaches modulating the tumor microenvironment, would be a promising alternative to improve immunotherapy efficacy in solid tumors.

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“…All described genome editing technologies have been employed for the modification of either hematopoietic stem cells or T cells. As summarized in Figure 3, genome editing has been exploited with several purposes in the ACT field: to completely redirect T cell specificity (42, 117, 124, 158-160), to avoid the risk of GvHD or fratricide effects mediated by CAR-T cells (161)(162)(163)(164)(165), to make adoptively transferred T cells resistant to the immunosuppressive environment (42, [166][167][168][169][170][171][172][173] or to lymphodepleting drugs (174,175). The high efficiency of gene editing and the overall flexibility of the CRISPR/Cas9 system makes it the most relevant tool to precisely and rapidly edit high numbers of T cells to be used in ACT.…”

Section: Genome Editing In the Service Of Actmentioning

confidence: 99%

TCR Redirected T Cells for Cancer Treatment: Achievements, Hurdles, and Goals

et al. 2020

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Adoptive T cell therapy (ACT) is a rapidly evolving therapeutic approach designed to harness T cell specificity and function to fight diseases. Based on the evidence that T lymphocytes can mediate a potent anti-tumor response, initially ACT solely relied on the isolation, in vitro expansion, and infusion of tumor-infiltrating or circulating tumor-specific T cells. Although effective in a subset of cases, in the first ACT clinical trials several patients experienced disease progression, in some cases after temporary disease control. This evidence prompted researchers to improve ACT products by taking advantage of the continuously evolving gene engineering field and by improving manufacturing protocols, to enable the generation of effective and long-term persisting tumor-specific T cell products. Despite recent advances, several challenges, including prioritization of antigen targets, identification, and optimization of tumor-specific T cell receptors, in the development of tools enabling T cells to counteract the immunosuppressive tumor microenvironment, still need to be faced. This review aims at summarizing the major achievements, hurdles and possible solutions designed to improve the ACT efficacy and safety profile in the context of liquid and solid tumors.

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CRISPR knock out of programmed cell death protein 1 enhances anti-tumor activity of cytotoxic T lymphocytes

Cited by 81 publications

References 19 publications

Repurposing endogenous immune pathways to tailor and control chimeric antigen receptor T cell functionality

Repurposing endogenous immune pathways to tailor and control chimeric antigen receptor T cell functionality

Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytes

TCR Redirected T Cells for Cancer Treatment: Achievements, Hurdles, and Goals

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