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

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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.

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Acknowledgements

This work was supported by a Breast Cancer Campaign Project Grant (2006NovPR18), the Guy’s and St Thomas’ Charity, Experimental Cancer Medicine Centre (King’s College London) and by Guy’s and the Department of Health via the National Institute for Health Research (NIHR) comprehensive Biomedical Research Centre (BRC) award to Guy’s & St Thomas’ NHS Foundation Trust in partnership with King’s College London and King’s College Hospital NHS Foundation Trust. CB is funded by the Oracle Cancer Trust at the Royal Marsden Hospital. SAE’s research is funded by the Institute of Cancer Research (ICR) and Cancer Research UK grant CA309/A8274 and funding provided via the NIHR specialist BRC award to the Royal Marsden NHS Foundation Trust in partnership with the ICR. We thank colleagues who have generously provided materials that facilitated this work, as indicated in the text.

Conflict of Interest Statement

The authors declare that they have no conflict of interest.

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Correspondence to John Maher.

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Fig S1

Western blotting analysis of ErbB expression. The indicated tumor cell lines were analyzed for expression of ErbB1-4 expression by western blotting under reducing conditions, using rabbit anti-ErbB antiserum. A cross-reactive band in the ErbB4 blot serves as a loading/transfer control. (JPEG 40 kb)

High resolution image (TIFF 655 kb)

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Wilkie, S., van Schalkwyk, M.C.I., Hobbs, S. et al. Dual Targeting of ErbB2 and MUC1 in Breast Cancer Using Chimeric Antigen Receptors Engineered to Provide Complementary Signaling. J Clin Immunol 32, 1059–1070 (2012). https://doi.org/10.1007/s10875-012-9689-9

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  • DOI: https://doi.org/10.1007/s10875-012-9689-9

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