Preclinical Strategies to Identify Off-Target Toxicity of High-Affinity TCRs

Helena M. Bijen, Dirk M. van der Steen, Renate S. Hagedoorn, Anne K. Wouters, Linda Wooldridge, J. H.Frederik Falkenburg, Mirjam H.M. Heemskerk*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

10 Citations (Scopus)
203 Downloads (Pure)

Abstract

Adoptive transfer of T cells engineered with a cancer-specific T cell receptor (TCR) has demonstrated clinical benefit. However, the risk for off-target toxicity of TCRs remains a concern. Here, we examined the cross-reactive profile of T cell clone (7B5) with a high functional sensitivity for the hematopoietic-restricted minor histocompatibility antigen HA-2 in the context of HLA-A*02:01. HA-2pos Epstein-Barr virus-transformed B lymphoblastic cell lines (EBV-LCLs) and primary acute myeloid leukemia samples, but not hematopoietic HA-2neg samples, are effectively recognized. However, we found unexpected off-target recognition of human fibroblasts and keratinocytes not expressing the HA-2 antigen. To uncover the origin of this off-target recognition, we performed an alanine scanning approach, identifying six out of nine positions to be important for peptide recognition. This indicates a low risk for broad cross-reactivity. However, using a combinatorial peptide library scanning approach, we identified a CDH13-derived peptide activating the 7B5 T cell clone. This was confirmed by recognition of CDH13-transduced EBV-LCLs and cell subsets endogenously expressing CDH13, such as proximal tubular epithelial cells. As such, we recommend the use of a combinatorial peptide library scan followed by screening against additional cell subsets to validate TCR specificity and detect off-target toxicity due to cross-reactivity directed against unrelated peptides before selecting candidate TCRs for clinical testing. Adoptive transfer of T cells engineered with a cancer-specific T cell receptor (TCR) has demonstrated clinical benefit. To validate TCR specificity and detect off-target toxicity before selecting candidate TCRs for clinical testing, Bijen et al. recommend the use of a combinatorial peptide library scan followed by screening against additional cell subsets.

Original languageEnglish
Pages (from-to)1206-1214
Number of pages9
JournalMolecular Therapy
Volume26
Issue number5
Early online date22 Feb 2018
DOIs
Publication statusPublished - 2 May 2018

Keywords

  • adoptive T cell therapy
  • off-target toxicity
  • preclinical screening
  • SCT
  • TCR

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