Anti-CD7 (3A1E) h(4-1BB-CD3ζ) CAR, pCDCAR1 (CAR-0120ZP3028)

CAR Construction : 3A1e-CD28-CD3ζ

Fig.1 Expanded CD7KO CD7 CAR T cells eradicate T-ALL and T lymphoma cell lines.

Absolute counts of live tumor cells were measured by flow cytometry using counting beads at the end of coculture with CD7KO CD7 CAR T cells.

Gomes-Silva, D., Srinivasan, M., Sharma, S., Lee, C. M., Wagner, D. L., Davis, T. H., ... & Mamonkin, M. (2017). CD7-edited T cells expressing a CD7-specific CAR for the therapy of T-cell malignancies. Blood, The Journal of the American Society of Hematology, 130(3), 285-296.

CAR Construction : 3A1e-CD28-CD3ζ

Fig.2 CD7KO CD7 CAR T cells eradicate AML Cell Lines.

Absolute counts of live AML cells were measured by flow cytometry using counting beads at the end of coculture with CD7KO CD7 CAR T cells.

Gomes-Silva, D., Atilla, E., Atilla, P. A., Mo, F., Tashiro, H., Srinivasan, M., ... & Mamonkin, M. (2019). CD7 CAR T cells for the therapy of acute myeloid leukemia. Molecular Therapy, 27(1), 272-280.

CAR Construction : 3A1e-CD28-CD3ζ

Fig.3 Determination of cytokines release.

Representative dot plots showing intracellular cytokine staining for TNFα and IFNγ in CD7KO CD7 CAR T cells upon coculture with CCRF cells or media alone.

Gomes-Silva, D., Srinivasan, M., Sharma, S., Lee, C. M., Wagner, D. L., Davis, T. H., ... & Mamonkin, M. (2017). CD7-edited T cells expressing a CD7-specific CAR for the therapy of T-cell malignancies. Blood, The Journal of the American Society of Hematology, 130(3), 285-296.

CAR Construction : 3A1e-CD28-CD3ζ

Fig.4 Determination of IFNγ production.

IFNγ production by CD8+and CD4+CD7 CAR T cells upon 4-hr coculture with indicated AML cell lines by intracellular cytokine staining and flow cytometry.

Gomes-Silva, D., Atilla, E., Atilla, P. A., Mo, F., Tashiro, H., Srinivasan, M., ... & Mamonkin, M. (2019). CD7 CAR T cells for the therapy of acute myeloid leukemia. Molecular Therapy, 27(1), 272-280.

CAR Construction : 3A1e-CD28-CD3ζ

Fig.5 CD7KO CD7 CAR T cells control the progression of systemic T-ALL in the mouse xenograft model.

NSG mice were injected intravenously with GFP-FFluc CCRF cells followed by a single intravenous injection of control or CD7KO CD7 CAR T cells 3 days later.

Gomes-Silva, D., Srinivasan, M., Sharma, S., Lee, C. M., Wagner, D. L., Davis, T. H., ... & Mamonkin, M. (2017). CD7-edited T cells expressing a CD7-specific CAR for the therapy of T-cell malignancies. Blood, The Journal of the American Society of Hematology, 130(3), 285-296.

CAR Construction : 3A1e-CD28-CD3ζ

Fig.6 CD7 CAR T cells control the progression of systemic AML in vivo.

NSG mice received FFluc-expressing KG-1a cells 24 hr after sublethal irradiation with 116 cGy. Eight days later, mice received a single injection of control or CD7 CAR T cells intravenously and were monitored for tumor progression.

Gomes-Silva, D., Atilla, E., Atilla, P. A., Mo, F., Tashiro, H., Srinivasan, M., ... & Mamonkin, M. (2019). CD7 CAR T cells for the therapy of acute myeloid leukemia. Molecular Therapy, 27(1), 272-280.

CAR Construction : 3A1e-41BB-CD3ζ

Fig.7 In vitro cytotoxicity of base edited CD3CAR and CD7CAR cells.

51Cr labelled Jurkat T cells modified to express CD3+CD7+, CD3+CD7−, CD3−CD7+ or CD3−CD7− were co-cultured with either CD3CAR , CD7CAR , mixed CD3CAR/CD7CAR or untransduced cells at an increasing ratio of effectors:targets (E:T).

Georgiadis, C., Rasaiyaah, J., Gkazi, S. A., Preece, R., Etuk, A., Christi, A., & Qasim, W. (2021). Base-edited CAR T cells for combinational therapy against T cell malignancies. Leukemia, 35(12), 3466-3481.

CAR Construction : 3A1e-41BB-CD3ζ

Fig.8 CD3CAR and CD7CAR T cells exhibit potent anti-leukaemic effects in vivo.

CD3CAR or CD7CAR cells edited using either SpCas9 or coBE3 mRNA were infused intravenously on day 4 alongside PBS and untransduced controls and leukaemic progression was monitored by serial bioluminescent imaging on over four weeks.

Georgiadis, C., Rasaiyaah, J., Gkazi, S. A., Preece, R., Etuk, A., Christi, A., & Qasim, W. (2021). Base-edited CAR T cells for combinational therapy against T cell malignancies. Leukemia, 35(12), 3466-3481.

CAR Construction : 3A1e-OX40-CD28-CD3ζ

Fig.9 Cell apoptosis analysis.

Cell apoptosis results revealed that the proportions of both early and late apoptotic cells in the anti-CD7 CAR-T cell groups were higher than those in the mock group and the anti-CD19 CAR-T cell group. Anti-CD7 CAR-T cells may induce fratricide because of T cell expression of CD7.

Ye, J., Jia, Y., Tuhin, I. J., Tan, J., Monty, M. A., Xu, N., ... & Yu, L. (2022). Feasibility study of a novel preparation strategy for anti-CD7 CAR-T cells with a recombinant anti-CD7 blocking antibody. Molecular Therapy-Oncolytics, 24, 719-728.

CAR Construction : 3A1e-OX40-CD28-CD3ζ

Fig.10 Anti-CD7 CAR-T cells cultured with an antibody possess strong cytotoxicity against CD7-positive tumor cells.

The cytotoxicity of nontransduced T cells (mock) and anti-CD7 CAR-T cells cultured with the PUT644 antibody against target Jurkat cells was evaluated.

Ye, J., Jia, Y., Tuhin, I. J., Tan, J., Monty, M. A., Xu, N., ... & Yu, L. (2022). Feasibility study of a novel preparation strategy for anti-CD7 CAR-T cells with a recombinant anti-CD7 blocking antibody. Molecular Therapy-Oncolytics, 24, 719-728.

CAR Construction :

Fig.11 The binding affinity measurement of 3Ale scfv and Fab to CD7-Rg.

Representative sensorgrams depicting the binding of 3Ale sFv (B), 3Ale Fab (D) to CD7-Rg (470 RU) were generated using a BlAcore instrument.

Pauza, M. E., Doumbia, S. O., & Pennell, C. A. (1997). Construction and characterization of human CD7-specific single-chain Fv immunotoxins. The Journal of Immunology, 158(7), 3259-3269.