Clinical Scale Zinc Finger Nuclease-mediated Gene Editing of PD-1 in Tumor Infiltrating Lymphocytes for the Treatment of Metastatic Melanoma.
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.
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA; Indiana University School of Medicine, Department of Surgery, Indianapolis, Indiana, USA.
Joal D
JD
Beane
Sangamo BioSciences, Richmond, California, USA.
Gary
G
Lee
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA.
Zhili
Z
Zheng
Sangamo BioSciences, Richmond, California, USA.
Matthew
M
Mendel
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA; Present Address: Moffitt Cancer Center, Tampa, FL.
Daniel
D
Abate-Daga
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA.
Mini
M
Bharathan
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA.
Mary
M
Black
Sangamo BioSciences, Richmond, California, USA.
Nimisha
N
Gandhi
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA.
Zhiya
Z
Yu
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA.
Smita
S
Chandran
Sangamo BioSciences, Richmond, California, USA.
Martin
M
Giedlin
Sangamo BioSciences, Richmond, California, USA.
Dale
D
Ando
Sangamo BioSciences, Richmond, California, USA.
Jeff
J
Miller
Sangamo BioSciences, Richmond, California, USA.
David
D
Paschon
Sangamo BioSciences, Richmond, California, USA.
Dmitry
D
Guschin
Sangamo BioSciences, Richmond, California, USA.
Edward J
EJ
Rebar
Sangamo BioSciences, Richmond, California, USA.
Andreas
A
Reik
Sangamo BioSciences, Richmond, California, USA.
Michael C
MC
Holmes
Sangamo BioSciences, Richmond, California, USA.
Philip D
PD
Gregory
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA.
Nicholas P
NP
Restifo
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA.
Steven A
SA
Rosenberg
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA; Present Address: Bluebird Bio, Cambridge, Massachusetts, USA.
Richard A
RA
Morgan
National Cancer Institute, Surgery Branch, Bethesda, Maryland, USA. Electronic address: feldmanst@mail.nih.gov.
Steven A
SA
Feldman
D000483:Alleles; D000818:Animals; D002469:Cell Separation; D016207:Cytokines / Q000378:metabolism; D004722:Endoribonucleases / Q000235:genetics*; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007156:Immunologic Memory; D016219:Immunotherapy, Adoptive; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology*; D008545:Melanoma / Q000628:therapy*; D051379:Mice; D009362:Neoplasm Metastasis; D009368:Neoplasm Transplantation; D010641:Phenotype; D061026:Programmed Cell Death 1 Receptor / Q000235:genetics* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D016335:Zinc Fingers
D016207:Cytokines; C105992:PDCD1 protein, human; D061026:Programmed Cell Death 1 Receptor; D014409:Tumor Necrosis Factor-alpha; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004722:Endoribonucleases