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Mol Ther. 2015 Aug;23(8):1380-1390. doi: 10.1038/mt.2015.71. Epub 2015 May 5.

Clinical Scale Zinc Finger Nuclease-mediated Gene Editing of PD-1 in Tumor Infiltrating Lymphocytes for the Treatment of Metastatic Melanoma.

Author information

1
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA; Indiana University School of Medicine, Department of Surgery, Indianapolis, Indiana, USA.
2
Sangamo BioSciences, Richmond, California, USA.
3
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA.
4
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA; Present Address: Moffitt Cancer Center, Tampa, FL.
5
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA; Present Address: Bluebird Bio, Cambridge, Massachusetts, USA.
6
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA. Electronic address: feldmanst@mail.nih.gov.

Abstract

Programmed cell death-1 (PD-1) is expressed on activated T cells and represents an attractive target for gene-editing of tumor targeted T cells prior to adoptive cell transfer (ACT). We used zinc finger nucleases (ZFNs) directed against the gene encoding human PD-1 (PDCD-1) to gene-edit melanoma tumor infiltrating lymphocytes (TIL). We show that our clinical scale TIL production process yielded efficient modification of the PD-1 gene locus, with an average modification frequency of 74.8% (n = 3, range 69.9-84.1%) of the alleles in a bulk TIL population, which resulted in a 76% reduction in PD-1 surface-expression. Forty to 48% of PD-1 gene-edited cells had biallelic PD-1 modification. Importantly, the PD-1 gene-edited TIL product showed improved in vitro effector function and a significantly increased polyfunctional cytokine profile (TNFα, GM-CSF, and IFNγ) compared to unmodified TIL in two of the three donors tested. In addition, all donor cells displayed an effector memory phenotype and expanded approximately 500-2,000-fold in vitro. Thus, further study to determine the efficiency and safety of adoptive cell transfer using PD-1 gene-edited TIL for the treatment of metastatic melanoma is warranted.

PMID:
25939491
PMCID:
PMC4817870
DOI:
10.1038/mt.2015.71
[Indexed for MEDLINE]
Free PMC Article

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