Gene therapy using genetically modified lymphocytes targeting VEGFR-2 inhibits the growth of vascularized syngenic tumors in mice
Dhanalakshmi Chinnasamy, Zhiya Yu, Marc R. Theoret, Yangbing Zhao, Rajeev K. Shrimali, Richard A. Morgan, Steven A. Feldman, Nicholas P. Restifo, Steven A. Rosenberg
Dhanalakshmi Chinnasamy, Zhiya Yu, Marc R. Theoret, Yangbing Zhao, Rajeev K. Shrimali, Richard A. Morgan, Steven A. Feldman, Nicholas P. Restifo, Steven A. Rosenberg
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Research Article Genetics

Gene therapy using genetically modified lymphocytes targeting VEGFR-2 inhibits the growth of vascularized syngenic tumors in mice

Abstract

Immunotherapies based on adoptive cell transfer are highly effective in the treatment of metastatic melanoma, but the use of this approach in other cancer histologies has been hampered by the identification of appropriate target molecules. Immunologic approaches targeting tumor vasculature provide a means for the therapy of multiple solid tumor types. We developed a method to target tumor vasculature, using genetically redirected syngeneic or autologous T cells. Mouse and human T cells were engineered to express a chimeric antigen receptor (CAR) targeted against VEGFR-2, which is overexpressed in tumor vasculature and is responsible for VEGF-mediated tumor progression and metastasis. Mouse and human T cells expressing the relevant VEGFR-2 CARs mediated specific immune responses against VEGFR-2 protein as well as VEGFR-2–expressing cells in vitro. A single dose of VEGFR-2 CAR-engineered mouse T cells plus exogenous IL-2 significantly inhibited the growth of 5 different types of established, vascularized syngeneic tumors in 2 different strains of mice and prolonged the survival of mice. T cells transduced with VEGFR-2 CAR showed durable and increased tumor infiltration, correlating with their antitumor effect. This approach provides a potential method for the gene therapy of a variety of human cancers.

Authors

Dhanalakshmi Chinnasamy, Zhiya Yu, Marc R. Theoret, Yangbing Zhao, Rajeev K. Shrimali, Richard A. Morgan, Steven A. Feldman, Nicholas P. Restifo, Steven A. Rosenberg

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Figure 2

DC101-CAR–transduced mouse T cells are functionally competent in generating immune responses against VEGFR-2–expressing mouse cell lines.

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DC101-CAR–transduced mouse T cells are functionally competent in generat...
(A) Expression of VEGFR-2 on mouse endothelial cells and tumor cells. Indicated mouse cell lines were incubated sequentially with recombinant DC101 antibody (a rat IgG antibody specific to mouse VEGFR-2) or isotype control rat IgG1, soluble mouse VEGFR-2–hIgG-Fc protein, and PE-labeled anti-hIgG-Fc (α-hIgG.Fc) and analyzed by FACS. Filled histogram, VEGFR-2–specific staining; open histogram, staining with rat IgG1. Results shown are representative of 2 experiments. (B) Primary mouse T cells were transduced with various retroviral vector constructs shown in Figure 1A. Four days later, cells were cocultured with the indicated mouse cell lines. Culture supernatants harvested at 24 hours after coculture were assayed for IFN-γ by ELISA. Results are presented as the mean of triplicate wells. Correlation between the IFN-γ secretion of DC101-CAR–engineered effector T cells and the MFI of VEGFR-2 expression on the target cells is shown (inset).