Highly efficient PD-1-targeted CRISPR-Cas9 for tumor-infiltrating lymphocyte-based adoptive T cell therapy

Christopher Aled Chamberlain*, Eric Paul Bennett, Anders Handrup Kverneland, Inge Marie Svane, Marco Donia, Özcan Met

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Adoptive T cell therapy (ACT) with expanded tumor-infiltrating lymphocytes (TIL) can induce durable responses in cancer patients from multiple histologies, with response rates of up to 50%. Antibodies blocking the engagement of the inhibitory receptor programmed cell death protein 1 (PD-1) have been successful across a variety of cancer diagnoses. We hypothesized that these approaches could be combined by using CRISPR-Cas9 gene editing to knock out PD-1 in TILs from metastatic melanoma and head-and-neck, thyroid, and colorectal cancer. Non-viral, non-plasmid-based PD-1 knockout was carried out immediately prior to the traditional 14-day TIL-based ACT rapid-expansion protocol. A median 87.53% reduction in cell surface PD-1 expression was observed post-expansion and confirmed at the genomic level. No off-target editing was detected, and PD-1 knockout had no effect on final fold expansion. Edited cells exhibited few phenotypic differences and matched control functionality. Pre-clinical-scale results were confirmed at a clinical scale by generating a PD-1-deficient TIL product using the good manufacturing practice facilities, equipment, procedures, and starting material used for standard patient treatment. Our results demonstrate that simple, non-viral, non-plasmid-based CRISPR-Cas9 methods can be feasibly adopted into a TIL-based ACT protocol to produce treatment products deficient in molecules such as PD-1, without any evident negative effects.

Original languageEnglish
JournalMolecular Therapy - Oncolytics
Volume24
Pages (from-to)417-428
Number of pages12
DOIs
Publication statusPublished - 2022

Bibliographical note

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© 2022 The Authors

Keywords

  • adoptive cell therapy
  • CRISPR-Cas9
  • gene editing
  • immunotherapy
  • PD-1
  • TIL therapy
  • tumor-infiltrating lymphocytes

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