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Sleeping Beauty (SB) transposon, a type of nonviral integrative vectors, provides an alternative to modify primary T cells. Creative biolabs has developed SB transposon CAR vector pSBCAR1 EphA2 (4H5) h(ICOSBBζ), which is constructed for the engineering of T cells to target human EphA2. The T cells are genetically modified through transduction with a nonviral vector expressing scFv of anti-EphA2 antibody linked to ICOS (CD278) signaling domain and CD137 (4-1BB), CD3-zeta signaling domains. And the vector product was designed for the treatment of Breast cancer.
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CAR Construction : 4H5 scFv-41BB-CD28ζ
Fig.1 Erythropoietin-producing hepatocellular carcinoma A2 (EphA2)-specific T cells recognize and kill EphA2-positive gliomas. Nontransduced (NT) or EphA2-specific T cells were cocultured with target cells at a 1:5 ratio, and after 24 hours, the production of interferon-γ (IFN-γ) and interleukin- 2 (IL-2) by T cells was determined by ELISA. Chow, K. K., Naik, S., Kakarla, S., Brawley, V. S., Shaffer, D. R., Yi, Z., Rainusso, N., Wu, M. F., Liu, H., Kew, Y., Grossman, R. G., Powell, S., Lee, D., Ahmed, N., & Gottschalk, S. (2013). T cells redirected to EphA2 for the immunotherapy of glioblastoma. Molecular therapy : the journal of the American Society of Gene Therapy, 21(3), 629–637. |
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CAR Construction : 4H5 scFv-41BB-CD28ζ
Fig.2 Cytotoxicity assays. Cytotoxicity assays with EphA2-specific T cells and NT-T cells as effectors and five primary EphA2-positive glioblastoma cell lines as targets. Chow, K. K., Naik, S., Kakarla, S., Brawley, V. S., Shaffer, D. R., Yi, Z., Rainusso, N., Wu, M. F., Liu, H., Kew, Y., Grossman, R. G., Powell, S., Lee, D., Ahmed, N., & Gottschalk, S. (2013). T cells redirected to EphA2 for the immunotherapy of glioblastoma. Molecular therapy : the journal of the American Society of Gene Therapy, 21(3), 629–637. |
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CAR Construction : 4H5 scFv-41BB-CD28ζ
Fig.3 Bioluminescence imaging. Bioluminescence imaging was used to follow tumor progression. All mice had detectable tumors just prior to treatment (day 7). By day 40, all mice in the control groups had progressive tumors (9/9 for untreated, 8/8 for NT-T cells). The tumor signal decreased in all mice treated with EphA2-specific T cells, and 50% (6/12) were long-term survivors. Chow, K. K., Naik, S., Kakarla, S., Brawley, V. S., Shaffer, D. R., Yi, Z., Rainusso, N., Wu, M. F., Liu, H., Kew, Y., Grossman, R. G., Powell, S., Lee, D., Ahmed, N., & Gottschalk, S. (2013). T cells redirected to EphA2 for the immunotherapy of glioblastoma. Molecular therapy : the journal of the American Society of Gene Therapy, 21(3), 629–637. |
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