Engineered T cells targeting E7 mediate regression of human papillomavirus cancers in a murine model
Benjamin Y. Jin, … , Cornelia L. Trimble, Christian S. Hinrichs
Benjamin Y. Jin, … , Cornelia L. Trimble, Christian S. Hinrichs
Published April 19, 2018
Citation Information: JCI Insight. 2018;3(8):e99488. https://doi.org/10.1172/jci.insight.99488.
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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.