Dual Targeting of ErbB2 and MUC1 in Breast Cancer Using Chimeric Antigen Receptors Engineered to Provide Complementary Signaling

J Clin Immunol. 2012 Oct;32(5):1059-70. doi: 10.1007/s10875-012-9689-9. Epub 2012 Apr 17.

Abstract

Purpose: Chimeric antigen receptor (CAR) engineered T-cells occupy an increasing niche in cancer immunotherapy. In this context, CAR-mediated CD3ζ signaling is sufficient to elicit cytotoxicity and interferon-γ production while the additional provision of CD28-mediated signal 2 promotes T-cell proliferation and interleukin (IL)-2 production. This compartmentalisation of signaling opens the possibility that complementary CARs could be used to focus T-cell activation within the tumor microenvironment.

Methods: Here, we have tested this principle by co-expressing an ErbB2- and MUC1-specific CAR that signal using CD3ζ and CD28 respectively. Stoichiometric co-expression of transgenes was achieved using the SFG retroviral vector containing an intervening Thosea asigna peptide.

Results: We found that "dual-targeted" T-cells kill ErbB2(+) tumor cells efficiently and proliferate in a manner that requires co-expression of MUC1 and ErbB2 by target cells. Notably, however, IL-2 production was modest when compared to control CAR-engineered T-cells in which signaling is delivered by a fused CD28 + CD3ζ endodomain.

Conclusions: These findings demonstrate the principle that dual targeting may be achieved using genetically targeted T-cells and pave the way for testing of this strategy in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / immunology*
  • Breast Neoplasms / immunology*
  • Cell Line, Tumor
  • Cell Proliferation
  • Cells, Cultured
  • Female
  • Humans
  • Immunotherapy, Adoptive*
  • Interferon-gamma / immunology
  • Interleukin-2 / immunology
  • Mucin-1 / immunology*
  • Receptors, Antigen, T-Cell / immunology*
  • Signal Transduction / immunology
  • T-Lymphocytes / immunology*

Substances

  • Adaptor Proteins, Signal Transducing
  • ERBIN protein, human
  • Interleukin-2
  • MUC1 protein, human
  • Mucin-1
  • Receptors, Antigen, T-Cell
  • Interferon-gamma