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 9

Recognition of KDR-expressing primary human cells by KDR-CAR–transduced PBLs.

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Recognition of KDR-expressing primary human cells by KDR-CAR–transduced ...
(A) KDR expression on various normal primary human cells. The indicated human cell types were stained with PE-labeled mouse anti-human KDR antibody or isotype control antibody and analyzed by FACS. The filled histograms indicate KDR-specific staining; the open histograms indicate staining with isotype control antibody. (B) Primary human T cells were mock transduced or retrovirally engineered to express KDR1121-hCD828BBZ CAR and, 8 days later, cocultured with indicated human cells for 24 hours. Culture supernatants were assayed for IFN-γ by ELISA. Results are presented as mean ± SEM of triplicates. (C) The cytotoxicity of genetically engineered primary human T cells expressing KDR1121-hCD828BBZ or KDR1121-hCD28Z CAR against indicated target cells was determined by using a standard Cr51 release assay. Each data point reflects the mean of triplicates.