Avoidance of On-Target Off-Tumor Activation Using a Co-stimulation-Only Chimeric Antigen Receptor

Mol Ther. 2017 May 3;25(5):1234-1247. doi: 10.1016/j.ymthe.2017.03.002. Epub 2017 Mar 22.

Abstract

Chimeric antigen receptors (CARs) combine T cell activation with antibody-mediated tumor antigen specificity, bypassing the need for T cell receptor (TCR) ligation. A limitation of CAR technology is on-target off-tumor toxicity caused by target antigen expression on normal cells. Using GD2 as a model cancer antigen, we hypothesized that this could be minimized by using T cells expressing Vγ9Vδ2 TCR, which recognizes transformed cells in a major histocompatibility complex (MHC)-unrestricted manner, in combination with a co-stimulatory CAR that would function independently of the TCR. An anti-GD2 CAR containing a solitary endodomain derived from the NKG2D adaptor DAP10 was expressed in Vγ9Vδ2+ T cells. Differential ligation of the CAR and/or TCR using antibody-coated beads showed that pro-inflammatory cytokine response depended on activation of both receptors. Moreover, in killing assays, GD2-expressing neuroblastoma cells that engaged the Vγ9Vδ2 TCR were efficiently lysed, whereas cells that expressed GD2 equivalently but did not engage the Vγ9Vδ2 TCR were untouched. Differentiation between X-on tumor and X-off tumor offers potential for safer immunotherapy and broader target selection.

Keywords: chimeric antigen receptor; toxicity; γδT cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Neoplasm / genetics*
  • Antigens, Neoplasm / immunology
  • CD28 Antigens / genetics
  • CD28 Antigens / immunology
  • CD3 Complex / genetics
  • CD3 Complex / immunology
  • Cell Line, Tumor
  • Coculture Techniques
  • Cytotoxicity, Immunologic
  • Gangliosides / chemistry*
  • Gangliosides / immunology
  • Gene Expression
  • Humans
  • Immunotherapy / methods
  • Lymphocyte Activation
  • Mutant Chimeric Proteins / genetics*
  • Mutant Chimeric Proteins / immunology
  • Neurons / immunology
  • Neurons / pathology
  • Protein Engineering / methods
  • Receptors, Antigen, T-Cell / genetics*
  • Receptors, Antigen, T-Cell / immunology
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology*

Substances

  • Antigens, Neoplasm
  • CD28 Antigens
  • CD3 Complex
  • CD3 antigen, zeta chain
  • Gangliosides
  • Mutant Chimeric Proteins
  • Receptors, Antigen, T-Cell
  • ganglioside, GD2