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

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. RestifoSteven A. Rosenberg, Richard A. Morgan, Steven A. Feldman

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)1380-1390
Number of pages11
JournalMolecular Therapy
Volume23
Issue number8
DOIs
StatePublished - Aug 1 2015

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Pharmacology
  • Drug Discovery

Fingerprint

Dive into the research topics of 'Clinical Scale Zinc Finger Nuclease-mediated Gene Editing of PD-1 in Tumor Infiltrating Lymphocytes for the Treatment of Metastatic Melanoma'. Together they form a unique fingerprint.

Cite this