Clinical Scale Zinc Finger Nuclease-mediated Gene Editing of PD-1 in Tumor Infiltrating Lymphocytes for the Treatment of Metastatic Melanoma |
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Authors: | Joal D Beane Gary Lee Zhili Zheng Matthew Mendel Daniel Abate-Daga Mini Bharathan Mary Black Nimisha Gandhi Zhiya Yu Smita Chandran Martin Giedlin Dale Ando Jeff Miller David Paschon Dmitry Guschin Edward J Rebar Andreas Reik Michael C Holmes Philip D Gregory Nicholas P Restifo Steven A Rosenberg Richard A Morgan Steven A Feldman |
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Institution: | 1.National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA;2.Indiana University School of Medicine, Department of Surgery, Indianapolis, Indiana, USA;3.Sangamo BioSciences, Richmond, California, USA;4.Present Address: Moffitt Cancer Center, Tampa, FL;5.Present Address: Bluebird Bio, Cambridge, Massachusetts, USA |
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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. |
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