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.
View: Text | PDF
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

×

Figure 7

Enhanced tumor infiltration of adoptively transferred DC101-CAR–transduced T cells in mice bearing established B16-F10 tumor.

Options: View larger image (or click on image) Download as PowerPoint
Enhanced tumor infiltration of adoptively transferred DC101-CAR–transduc...
C57BL/6 mice bearing B16-F10 tumor were treated with 2 × 107 DC101-CD828BBZ CAR- or empty vector–transduced Thy1.1+ syngeneic T cells plus rhIL-2. Tumors and spleens from individual mice from each group were excised and processed to obtain single cell suspensions and analyzed by flow cytometry. (A) Representative FACS data from 3 experiments. (B) Pooled data obtained from 3 different mice from independent experiments (mean ± SEM). (C–E) Tumor samples were obtained from C57BL/6 mice bearing B16-F10 tumors treated with DC101-CD828BBZ CAR- or empty vector–transduced T cells and rhIL-2 on day 4 after ACT. Tumor sections were stained for Thy1.1 antigen expressed on transferred T cells (green) or the endothelial cell marker CD31 (red) together with DAPI (blue) to show the nucleus and analyzed using fluorescence microscopy. (C) Enhanced infiltration of adoptively transferred DC101-CD828BBZ CAR–transduced Thy1.1+ T cells into tumor (original magnification, ×10). (D) Different area of the same tumor section (original magnification, ×40) from a mouse treated with DC101-CAR–transduced T cells presented in C, showing infused Thy1.1+ T cells localized in and around the tumor vessels on day 4 after ACT. Yellow represents areas of colocalization of Thy1.1+ T cell (green) and endothelial cells (red). (E) CD31 (red) staining of tumor vessels in a representative section, showing reduction in vessels in the tumor of a mouse treated with DC101-CD828BBZ CAR-transduced T cells at day 6 after ACT (original magnification, ×10).