Gene therapy using genetically modified lymphocytes targeting VEGFR-2 inhibits the growth of vascularized syngenic tumors in mice
Dhanalakshmi Chinnasamy, … , Nicholas P. Restifo, Steven A. Rosenberg
Dhanalakshmi Chinnasamy, … , Nicholas P. Restifo, Steven A. Rosenberg
Published October 11, 2010
Citation Information: J Clin Invest. 2010;120(11):3953-3968. https://doi.org/10.1172/JCI43490.
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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 8

Construction and characterization of retroviral vectors encoding a CAR targeted against human VEGFR-2.

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Construction and characterization of retroviral vectors encoding a CAR t...
(A) KDR1121, a ScFv comprising the VL and VH of fully human IgG specific to human KDR antigen, fused by a 218-linker sequence (L). CD28, 4-1BB, and CD3-z intracellular T cell signaling domains derived from human CD28, TNFRSF9, and CD3 genes, respectively. LS, mouse immunoglobulin κ chain leader sequence; CD8, hinge and transmembrane regions from human CD8α. (B) Representative FACS data from 3 different donors transduced similarly with KDR1121-CD828Z or KDR1121-hCD28BBZ CAR-encoding retroviral vectors, showing the percentage of CD3+ T cells expressing KDR-CAR in the top right quadrants and the percentage of CD3+ T cells negative for KDR-CAR expression in the bottom right quadrants. (C and D) Primary human T cells were transduced with the indicated retroviral vectors. Seven days later, cells were cultured on antigen-coated 96-well microtiter plates for 3 days. (C) Cell proliferation was measured as [3H]thymidine incorporation during the last 16 hours. (D) Culture supernatants were assayed for IFN-γ by ELISA. Results are presented as the mean ± SEM of triplicates. (E) Primary human T cells from 3 different donors were mock transduced or transduced with KDR1121-hCD828BBZ CAR-expressing retroviral vector and, 8 days later, cocultured with the indicated cell lines for 24 hours. Culture supernatants were assayed for IFN-γ by ELISA.