Engineered T cells targeting E7 mediate regression of human papillomavirus cancers in a murine model
Benjamin Y. Jin, Tracy E. Campbell, Lindsey M. Draper, Sanja Stevanović, Bianca Weissbrich, Zhiya Yu, Nicholas P. Restifo, Steven A. Rosenberg, Cornelia L. Trimble, Christian S. Hinrichs
Benjamin Y. Jin, Tracy E. Campbell, Lindsey M. Draper, Sanja Stevanović, Bianca Weissbrich, Zhiya Yu, Nicholas P. Restifo, Steven A. Rosenberg, Cornelia L. Trimble, Christian S. Hinrichs
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Research Article Immunology

Engineered T cells targeting E7 mediate regression of human papillomavirus cancers in a murine model

Abstract

T cell receptor (TCR) T cell therapy is a promising cancer treatment modality. However, its successful development for epithelial cancers may depend on the identification of high-avidity TCRs directed against tumor-restricted target antigens. The human papillomavirus (HPV) E7 antigen is an attractive therapeutic target that is constitutively expressed by HPV+ cancers but not by healthy tissues. It is unknown if genetically engineered TCR T cells that target E7 can mediate regression of HPV+ cancers. We identified an HPV-16 E7-specific, HLA-A*02:01-restricted TCR from a uterine cervix biopsy from a woman with cervical intraepithelial neoplasia. This TCR demonstrated high functional avidity, with CD8 coreceptor–independent tumor targeting. Human T cells transduced to express the TCR specifically recognized and killed HPV-16+ cervical and oropharyngeal cancer cell lines and mediated regression of established HPV-16+ human cervical cancer tumors in a mouse model. These findings support the therapeutic potential of this approach and established the basis for an E7 TCR gene therapy clinical trial in patients with metastatic HPV+ cancers (NCT02858310).

Authors

Benjamin Y. Jin, Tracy E. Campbell, Lindsey M. Draper, Sanja Stevanović, Bianca Weissbrich, Zhiya Yu, Nicholas P. Restifo, Steven A. Rosenberg, Cornelia L. Trimble, Christian S. Hinrichs

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Figure 4

The avidity of E7 TCR T cells for cognate antigen and tumor cell lines.

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The avidity of E7 TCR T cells for cognate antigen and tumor cell lines. ...
(A) A functional avidity assay is shown with the quantity of IFN-γ produced in an overnight coculture graphed. The target cells are T2 cells pulsed with E629–38 or E711–19 peptide at the concentrations indicated on the x axis. The TCR/target antigen combinations are shown in the key. Error bars represent the SEM of 3 technical replicates. ***P < 0.001. (B) Koff rate assay evaluating the E6 and E7 TCRs. E7 TCR– or E6 TCR–transduced T cells were labeled with reversible, fluorescence-labeled MHC streptamers. The MHC fluorescence intensity of the transduced T cells after the addition of biotin was monitored by flow cytometry. Mean MHC fluorescence intensities of E6 TCR (triangles) and E7 TCR (squares) were extracted for every 8 seconds, normalized, and plotted to calculate the t1/2 time after fitting an exponential decay curve (7 independent dissociations per TCR, mean with SEM). (C) t1/2 time from 7 independent peptide-MHC dissociation experiments of the E6 TCR (triangles) and the E7 TCR (squares) are plotted with the median t1/2 time. ***P < 0.001. (D and E) Cytokine production assay testing the recognition of tumor cell lines by E6 and E7 TCR T cells. (D) CD4 and (E) CD8 T cells were isolated, transduced, and tested separately in functional assays. The quantity of IFN-γ produced in an overnight coculture of TCR-transduced T cells and target cells is shown. The target cells are CaSki, 4050, SCC152, SCC90, 624-E6/E7, and 624 cells. Error bars represent the SEM of 2 technical replicates. *P < 0.05, ***P < 0.001, ****P < 0.0001. The result shown is representative of 2 independent experiments. (F) CD4 or CD8 T cells were transduced to express the E6 or E7 TCR and were cocultured with 4050, CaSki, or 624 tumor cell lines. T cell–mediated cytolysis was monitored using the ACEA xCELLigence Real-Time Cell Analyzer. Cell indices plotted in the graphs represent the mean of duplicate samples, and error bars represent the SEM. The result shown is representative of 2 independent experiments. UT, untransduced T cells; E7 TCR, E7 TCR–transduced T cells; E6 TCR, E6 TCR–transduced T cells.