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CRISPR/Cas9 介导的 PD-1 敲除增强了嵌合抗原受体 T 细胞的抗肿瘤疗效。

CRISPR/Cas9-mediated PD-1 disruption enhances anti-tumor efficacy of human chimeric antigen receptor T cells.

机构信息

Department of Cellular & Molecular Pharmacology, San Francisco, CA, USA.

Center for Systems and Synthetic Biology, University of California San Francisco, San Francisco, 94158, CA, USA.

出版信息

Sci Rep. 2017 Apr 7;7(1):737. doi: 10.1038/s41598-017-00462-8.


DOI:10.1038/s41598-017-00462-8
PMID:28389661
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5428439/
Abstract

Immunotherapies with chimeric antigen receptor (CAR) T cells and checkpoint inhibitors (including antibodies that antagonize programmed cell death protein 1 [PD-1]) have both opened new avenues for cancer treatment, but the clinical potential of combined disruption of inhibitory checkpoints and CAR T cell therapy remains incompletely explored. Here we show that programmed death ligand 1 (PD-L1) expression on tumor cells can render human CAR T cells (anti-CD19 4-1BBζ) hypo-functional, resulting in impaired tumor clearance in a sub-cutaneous xenograft model. To overcome this suppressed anti-tumor response, we developed a protocol for combined Cas9 ribonucleoprotein (Cas9 RNP)-mediated gene editing and lentiviral transduction to generate PD-1 deficient anti-CD19 CAR T cells. Pdcd1 (PD-1) disruption augmented CAR T cell mediated killing of tumor cells in vitro and enhanced clearance of PD-L1+ tumor xenografts in vivo. This study demonstrates improved therapeutic efficacy of Cas9-edited CAR T cells and highlights the potential of precision genome engineering to enhance next-generation cell therapies.

摘要

嵌合抗原受体 (CAR) T 细胞和检查点抑制剂(包括拮抗程序性细胞死亡蛋白 1 [PD-1] 的抗体)的免疫疗法都为癌症治疗开辟了新途径,但联合破坏抑制性检查点和 CAR T 细胞疗法的临床潜力仍未得到充分探索。在这里,我们表明肿瘤细胞上的程序性死亡配体 1 (PD-L1) 的表达可以使人类 CAR T 细胞(抗-CD19 4-1BBζ)功能低下,导致皮下异种移植模型中的肿瘤清除受损。为了克服这种受抑制的抗肿瘤反应,我们开发了一种联合 Cas9 核糖核蛋白 (Cas9 RNP) 介导的基因编辑和慢病毒转导的方案,以生成 PD-1 缺失的抗-CD19 CAR T 细胞。Pdcd1(PD-1)的破坏增强了 CAR T 细胞对肿瘤细胞的体外杀伤作用,并增强了体内 PD-L1+肿瘤异种移植物的清除。这项研究证明了 Cas9 编辑的 CAR T 细胞的治疗效果得到了改善,并强调了精确基因组工程增强下一代细胞疗法的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2c0/5428439/225c065a0e39/41598_2017_462_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2c0/5428439/57341e6dc2d7/41598_2017_462_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2c0/5428439/a31c2caabed9/41598_2017_462_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2c0/5428439/225c065a0e39/41598_2017_462_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2c0/5428439/57341e6dc2d7/41598_2017_462_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2c0/5428439/a31c2caabed9/41598_2017_462_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b2c0/5428439/225c065a0e39/41598_2017_462_Fig3_HTML.jpg

相似文献

[1]
CRISPR/Cas9-mediated PD-1 disruption enhances anti-tumor efficacy of human chimeric antigen receptor T cells.

Sci Rep. 2017-4-7

[2]
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[3]
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Hum Gene Ther. 2018-4-27

[4]
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[5]
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Clin Cancer Res. 2021-1-15

[6]
CRISPR/Cas9-mediated PD-1 disruption enhances human mesothelin-targeted CAR T cell effector functions.

Cancer Immunol Immunother. 2018-12-6

[7]
PD-1 silencing improves anti-tumor activities of human mesothelin-targeted CAR T cells.

Hum Immunol. 2021-2

[8]
A versatile system for rapid multiplex genome-edited CAR T cell generation.

Oncotarget. 2017-3-7

[9]
Allogeneic chimeric antigen receptor-T cells with CRISPR-disrupted programmed death-1 checkpoint exhibit enhanced functional fitness.

Cytotherapy. 2023-7

[10]
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Nat Commun. 2024-4-26

引用本文的文献

[1]
Advancing gene editing therapeutics: Clinical trials and innovative delivery systems across diverse diseases.

Mol Ther Nucleic Acids. 2025-8-5

[2]
A Singular Base Editing Platform for Polyfunctional Multiplex Engineering of Immune Cells.

bioRxiv. 2025-7-16

[3]
Resistance Mechanisms to BCMA Targeting Bispecific Antibodies and CAR T-Cell Therapies in Multiple Myeloma.

Cells. 2025-7-15

[4]
CRISPR/Cas9 technology in tumor research and drug development application progress and future prospects.

Front Pharmacol. 2025-7-8

[5]
Sustained and specific multiplexed immune checkpoint modulation in CAR T cells induced by targeted epigenome editing.

Mol Ther Nucleic Acids. 2025-7-1

[6]
CRISPR/Cas9 for achieving postintervention HIV control.

Curr Opin HIV AIDS. 2025-9-1

[7]
Recent advances and challenges of cellular immunotherapies in lung cancer treatment.

Exp Hematol Oncol. 2025-7-7

[8]
CAR-T cell therapy in brain malignancies: obstacles in the face of cellular trafficking and persistence.

Front Immunol. 2025-6-19

[9]
CAR-T cell therapy for cancer: current challenges and future directions.

Signal Transduct Target Ther. 2025-7-4

[10]
Armored human CAR T cells with PD1 promoter-driven IL-10 have enhanced suppressive function.

Sci Adv. 2025-6-13

本文引用的文献

[1]
Early memory phenotypes drive T cell proliferation in patients with pediatric malignancies.

Sci Transl Med. 2016-1-6

[2]
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Nature. 2016-1-28

[3]
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Science. 2016-1-1

[4]
Structural Design of Engineered Costimulation Determines Tumor Rejection Kinetics and Persistence of CAR T Cells.

Cancer Cell. 2015-10-12

[5]
Remote control of therapeutic T cells through a small molecule-gated chimeric receptor.

Science. 2015-10-16

[6]
Melanoma Cell-Intrinsic PD-1 Receptor Functions Promote Tumor Growth.

Cell. 2015-9-10

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Metabolic Competition in the Tumor Microenvironment Is a Driver of Cancer Progression.

Cell. 2015-9-10

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Synthetic biology approaches to engineer T cells.

Curr Opin Immunol. 2015-8

[9]
Generation of knock-in primary human T cells using Cas9 ribonucleoproteins.

Proc Natl Acad Sci U S A. 2015-8-18

[10]
A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks.

Cell. 2015-7-30

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