Chimeric antigen receptor (CAR) T cell therapy is a powerful adoptive immunotherapy against blood cancers, but the therapeutic effect was not efficient enough on solid tumors. B cells have been reported to play a critical role in regulating memory T differentiation and cytotoxic T development. However, as of yet the influence of such B cells on CAR T cells has not been discussed. In this study, using ephrin type-A receptor 2 (EphA2) specific CAR T cells, we cultured B cells successfully to stimulate CAR T cells in vitro, and investigated the cell differentiation and anti-tumor efficiency. We observed that EphA2-CAR T cells stimulated by B cells performed increased interferon γ (IFN γ) production and upregulated OX40 expression, as well as the enhanced anti-tumor activity and reduced PD-1 expression. The persistence of CAR T cells was enhanced after B cells stimulation for more than 7 days with the increased subset of central memory T cells (T_CM). In addition, next generation sequencing was performed to explore the underlying mechanisms. The up-regulated genes clustered in, immune response activation, chemokine signaling pathway, calcium signaling pathway, cGMP-PKG signaling pathway and et al. which contributed to the upregulated anti-glioblastoma (GBM) activity of CAR T cells stimulated by B cell. Furthermore, MEF2C, CD40, SYK and TNFRSF13B were upregulated in CAR T cells after co-culturing with B cells. These genes functionally enriched in promoting lymphocytes proliferation and may contribute to the enhanced persistence of CAR T cells. In conclusion, these results indicated the critical role of B cells in prolonging CAR T cells longevity and enhancing anti-tumor activity, which paves the way for the therapeutic exploitation of EphA2-CAR T cells against GBM in the future.

2021-11-01·International Journal of Radiation Oncology*Biology*Physics

Development of Radioresistant CAR T Cells for Solid Tumor Therapeutics

Article

作者: Colgan, C ; DeSelm, C J ; Andruska, N

PURPOSE/OBJECTIVE(S)Chimeric antigen receptor T-cells (CAR-T) targeting CD19 have shown remarkable antitumor activity in relapsed/refractory hematological malignancies, but limited efficacy in solid tumors. Ionizing radiation can debulk tumors, thereby increasing CAR T cell penetration, and sensitize tumor cells to CAR-mediated killing. Thus, implementing CAR therapy within current RT regimens may further increase local and systemic tumor responses in patients. However, lymphocytes are highly radiosensitive, which can inhibit the use of radiation in conjunction with CAR T cell therapy. To circumvent this problem, we designed CAR T cells that when activated are more highly radioresistant.MATERIALS/METHODSEphA2-specitifc human CAR T cells were developed using a EphA2 scFv and CD28.ζ signaling domains (EphA2 CAR). These CAR T cells were further modified to include CAR signaling response elements upstream of the superoxide dismutase 2 gene (EphA2-SOD2 CAR), which neutralizes superoxide radicals. When EphA2-SOD2 CAR T cells are activated, they overexpress SOD2. CAR T cells were generated from healthy donors, cocultured with EphA2⁺ tumor cells for 3 days, treated with increasing doses of ionizing radiation (0-8 Gy), and compared in functional assay to measure the radiation sensitivity, specificity, and effector function of EphA2- and EphA2-SOD2 CAR T cells in vitro.RESULTSAt each dose of ionizing radiation, the surviving fraction of T lymphocytes was higher in the EphA2-SOD2 CAR T cells relative to the EphA2 CAR T cells. We performed cytotoxic T lymphocyte (CTL) assays in which EphA2⁺ H&N tumor cells that constitutively express firefly luciferase were cocultured for 24 hours with varying ratios of each CAR T cell. In the presence of survival cytokines, EphA2 CAR T cells and EphA2-SOD2 CAR T displayed similar killing capacity at different effector CAR T to tumor (E:T) ratios. However, in the absence of added survival cytokines, EphA2-SOD2 CAR T cells produced higher levels of tumor cell killing at each E:T ratio. EPhA2-SOD2 CAR T were unable to kill EphA2 knockout H&N tumor cells, demonstrating that EphA2-SOD2 CAR T cells specifically target EphA2⁺ tumor.CONCLUSIONEphA2-SOD2 CAR T are more radioresistant and exhibit higher killing capacity relative to standard CAR T cells. Radioresistant CAR T cells have promise and may further enhance the clinical utility of CAR T cells.

100 项与 EPHA2靶向CAR-T(泛恩生物) 相关的药物交易

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