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

• Published in: Journal of clinical immunology Vol. 32; no. 5; pp. 1059 - 1070
• Main Authors: Wilkie, Scott, van Schalkwyk, May C. I., Hobbs, Steve, Davies, David M., van der Stegen, Sjoukje J. C., Pereira, Ana C. Parente, Burbridge, Sophie E., Box, Carol, Eccles, Suzanne A., Maher, John
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.10.2012; Springer Nature B.V
• Subjects: Adaptor Proteins, Signal Transducing - immunology; Biomedical and Life Sciences; Biomedicine; Breast cancer; Breast Neoplasms - immunology; CD28 antigen; Cell Line, Tumor; Cell Proliferation; Cells, Cultured; Cytotoxicity; ErbB-2 protein; Female; Humans; Immunology; Immunotherapy; Immunotherapy, Adoptive; Infectious Diseases; Interferon-gamma - immunology; Interleukin 2; Interleukin-2 - immunology; Internal Medicine; Lymphocytes T; Medical Microbiology; Microenvironments; Mucin-1 - immunology; Niches; Receptors, Antigen, T-Cell - immunology; Signal Transduction - immunology; T-Lymphocytes - immunology; Transgenes; Tumor cells; targeting; MUC1; dual; ErbB2; Adoptive immunotherapy; specificity; chimeric antigen receptor
• ISSN: 0271-9142; 1573-2592
• DOI: 10.1007/s10875-012-9689-9

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.