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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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 6

Impact of exogenous rhIL-2, 4-1BB signaling, and host lymphodepletion on tumor treatment effect of DC101-CAR–transduced T cells.

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Impact of exogenous rhIL-2, 4-1BB signaling, and host lymphodepletion on...
(A) DC101-CAR–transduced T cells required exogenous rhIL-2 but not 4-1BB signaling for effective tumor treatment. C57BL/6 mice bearing subcutaneous B16-F10 tumors received 2 × 107 syngeneic T cells transduced with DC101-CAR containing the 4-1BB signaling domain (DC101-CD828BBZ, blue diamonds) or lacking 4-1BB (DC101-CD828Z; red triangles) or an empty vector (green squares). Control groups received no T cells (black circles). Groups represented by filled symbols received exogenous rhIL-2 administration, and those represented by open symbols did not receive rhIL-2. (B) 4-1BB signaling enhanced persistence of DC101-CAR–modified T cells in vivo. Tumor samples from 2 mice treated with DC101-CAR–transduced T cells plus rhIL-2, shown in Figure 4A, were harvested on day 30 after T cell transfer. The low-density cell fraction was prepared from tumor samples, and cell surface expression of DC101-CAR was determined by FACS. The percentage of CD3+ T cells expressing DC101-CAR in the lymphocyte-gated region of the forward and side scatter profiles is shown in top right quadrants, and the percentage of CD3+ T cells negative for DC101-CAR expression is shown in the bottom right quadrants. (C) Impact of host lymphodepletion on in vivo tumor therapeutic effect of DC101-CAR–engineered T cells. C57BL/6 mice bearing subcutaneous B16-F10 tumors received 2 × 107 syngeneic T cells transduced with DC101-CAR (red triangles), SP6-CAR (blue diamonds), or an empty vector (green squares) plus rhIL-2 or were not treated with T cells (black circles). Mice in groups represented by filled symbols received 5 Gy TBI prior to T cell transfer, and mice in groups represented by open symbols did not receive 5 Gy TBI. (A and C) Serial, blinded tumor measurements were obtained, and the products of perpendicular diameters were plotted ± SEM.