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

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

Rupp LJ1,2,3,4, Schumann K5,6, Roybal KT1,2,3, Gate RE7,8, Ye CJ7, Lim WA9,10,11,12, Marson A3,5,6,13,14.

Author information

1
Department of Cellular & Molecular Pharmacology, San Francisco, CA, USA.
2
Center for Systems and Synthetic Biology, University of California San Francisco, San Francisco, 94158, CA, USA.
3
Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, CA, 94158, USA.
4
Howard Hughes Medical Institute, University of California San Francisco, San Francisco, 94158, CA, USA.
5
Diabetes Center, University of California San Francisco, San Francisco, CA, 94143, USA.
6
Department of Microbiology and Immunology, University of California San Francisco, San Francisco, CA, 94143, USA.
7
Department of Epidemiology and Biostatistics, Department of Bioengineering and Therapeutic Sciences, Institute for Human Genetics, University of California, San Francisco, CA, 94143, USA.
8
Biological and Medical Informatics Graduate Program, University of California, San Francisco, CA, 94158, USA.
9
Department of Cellular & Molecular Pharmacology, San Francisco, CA, USA. Wendell.lim@ucsf.edu.
10
Center for Systems and Synthetic Biology, University of California San Francisco, San Francisco, 94158, CA, USA. Wendell.lim@ucsf.edu.
11
Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, CA, 94158, USA. Wendell.lim@ucsf.edu.
12
Howard Hughes Medical Institute, University of California San Francisco, San Francisco, 94158, CA, USA. Wendell.lim@ucsf.edu.
13
Division of Infectious Diseases and Rheumatology, Department of Medicine, University of California San Francisco, San Francisco, CA, 94143, USA.
14
Innovative Genomics Institute, University of California Berkeley, Berkeley, CA, 94720, USA.

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.

PMID:
28389661
PMCID:
PMC5428439
DOI:
10.1038/s41598-017-00462-8
[Indexed for MEDLINE]
Free PMC Article

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