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The vector of anti-CD38 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human CD38. The T cells are genetically modified through transduction with a retroviral vector expressing scFv of anti-CD38 antibody linked to 41BB and CD3ζ signaling domains. And the vector product was designed for the treatment of Multiple myeloma.
CAR Construction : 028-41BB-CD3ζ Fig.1 Efficacy of CD38-CART cells at lysing MM cell lines. In 24 h cytotoxicity assays, three different types of CD38-CART cells were tested against two MM cell lines with different CD38 expression levels: (Upper) U266, a CD38− cell line, (Bottom) UM9, a CD38+ cell line. Drent, E., Groen, R. W., Noort, W. A., Themeli, M., van Bueren, J. J. L., Parren, P. W., ... & Mutis, T. (2016). Pre-clinical evaluation of CD38 chimeric antigen receptor engineered T cells for the treatment of multiple myeloma. Haematologica, 101(5), 616. |
CAR Construction : 028-41BB-CD3ζ Fig.2 Cytokine release assay. 24 hr after co-incubation with the CD38+ target cell line UM9 or CD38 target U266, E:T ratio 1:1, cytokine secretion by mock or CD38-CAR028, A1, A4, B1, or B3 T cells was measured with a flow-cytometry-based assay in the cell-free supernatants. Drent, E., Themeli, M., Poels, R., de Jong-Korlaar, R., Yuan, H., de Bruijn, J., ... & Mutis, T. (2017). A rational strategy for reducing on-target off-tumor effects of CD38-chimeric antigen receptors by affinity optimization. Molecular therapy, 25(8), 1946-1958. |
CAR Construction : 028-41BB-CD3ζ Fig.3 CART cell phenotyping assay. Phenotypic profile of each CD38-CAR T cell type was determined before (week 0) and after (week 1) expansion with markers CD45RA and CD62L. Percentage of total cells is depicted for naive (CD45RA+/CD62L+), central memory (CM) (CD45RA/CD62L+), effector memory (EM) (CD45RA/CD62L), and effector(CD45RA+/CD62L). Drent, E., Themeli, M., Poels, R., de Jong-Korlaar, R., Yuan, H., de Bruijn, J., ... & Mutis, T. (2017). A rational strategy for reducing on-target off-tumor effects of CD38-chimeric antigen receptors by affinity optimization. Molecular therapy, 25(8), 1946-1958. |
CAR Construction : 028-41BB-CD3ζ Fig.4 In vivo efficacy of CD38-CAR-T cells. Mice were i.v. injected with 10* 10^6 cells of tumor cell line UM9 and treated 1 week after with i.v. injections of 5 * 10^6 mock, high-affinity CD38-CAR028, or low-affinity CD38-CARA4 T cells. Drent, E., Themeli, M., Poels, R., de Jong-Korlaar, R., Yuan, H., de Bruijn, J., ... & Mutis, T. (2017). A rational strategy for reducing on-target off-tumor effects of CD38-chimeric antigen receptors by affinity optimization. Molecular therapy, 25(8), 1946-1958. |
CAR Construction : 028-41BB-CD3ζ, 028-CD28-CD3ζ, 028-CD28-CD3ζ-41BBL Fig.5 The lytic capacity of CD38-CART cells. The series of high and low affinity CAR T cells were incubated with Firefly-Luciferase-transduced human MM cell line UM9. Drent, E., Poels, R., Ruiter, R., van de Donk, N. W., Zweegman, S., Yuan, H., ... & Themeli, M. (2019). Combined CD28 and 4-1BB Costimulation Potentiates Affinity-tuned Chimeric Antigen Receptor–engineered T CellsDual Costimulation Empowers Very Low–affinity CAR-T Cells. Clinical Cancer Research, 25(13), 4014-4025. |
CAR Construction : 028-41BB-CD3ζ, 028-CD28-CD3ζ, 028-CD28-CD3ζ-41BBL Fig.6 Cytokine release assay. 24 hours after co-incubation with UM9 (E:T ratio 1:1), cell supernatants were harvested to measure cytokine secretion with a flow cytometry-based assay. Graph shows the secretion of IFN-γ, TNF and IL-2. Drent, E., Poels, R., Ruiter, R., van de Donk, N. W., Zweegman, S., Yuan, H., ... & Themeli, M. (2019). Combined CD28 and 4-1BB Costimulation Potentiates Affinity-tuned Chimeric Antigen Receptor–engineered T CellsDual Costimulation Empowers Very Low–affinity CAR-T Cells. Clinical Cancer Research, 25(13), 4014-4025. |
CAR Construction : 028-41BB-CD3ζ, 028-CD28-CD3ζ, 028-CD28-CD3ζ-41BBL Fig.7 Exhaustion of CD38-CAR T cells. Pie charts illustrating the % of cells expressing either 0, 1 (PD-1+, Lag3+ or TIM3+), 2 (PD-1+/Lag3+ or PD-1+/TIM3+ or Lag3+/TIM3+) or 3 (PD-1+, Lag3+ and TIM3+) exhaustion markers gated on live CD3+CAR+ cells. Drent, E., Poels, R., Ruiter, R., van de Donk, N. W., Zweegman, S., Yuan, H., ... & Themeli, M. (2019). Combined CD28 and 4-1BB Costimulation Potentiates Affinity-tuned Chimeric Antigen Receptor–engineered T CellsDual Costimulation Empowers Very Low–affinity CAR-T Cells. Clinical Cancer Research, 25(13), 4014-4025. |
CAR Construction : Fig.8 Binding affinity of anti-CD38 scFv antibodies. Binding kinetics of anti-CD38 028 scFv antibodies against purified hCD38-ECD were investigated by biolayer interferometry. Ma, P., Ren, P., Zhang, C., Tang, J., Yu, Z., Zhu, X., ... & Lerner, R. A. (2021). Avidity‐Based Selection of Tissue‐Specific CAR‐T Cells from a Combinatorial Cellular Library of CARs. Advanced Science, 8(6), 2003091. |
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