Anti-human CD117 CAR T-cells efficiently eliminate healthy and malignant CD117-expressing hematopoietic cells


Acute myeloid leukemia (AML) initiating and sustaining cells maintain high cell-surface similarity with their cells-of-origin, i.e., hematopoietic stem and progenitor cells (HSPCs), and identification of truly distinguishing leukemia-private antigens has remained elusive to date. To nonetheless utilize surface antigen-directed immunotherapy in AML, we here propose targeting both, healthy and malignant human HSPC, by chimeric antigen receptor (CAR) T-cells with specificity against CD117, the cognate receptor for stem cell factor. This approach should spare most mature hematopoietic cells and would require CAR T termination followed by subsequent transplantation of healthy HSPCs to rescue hematopoiesis. We successfully generated anti-CD117 CAR T-cells from healthy donors and AML patients. Anti-CD117 CAR T-cells efficiently targeted healthy and leukemic CD117-positive cells in vitro. In mice xenografted with healthy human hematopoiesis, they eliminated CD117-expressing, but not CD117-negative human cells. Importantly, in mice xenografted with primary human CD117-positive AML, they eradicated disease in a therapeutic setting. Administration of ATG in combination with rituximab, which binds to the co-expressed CAR T-cell transduction/selection marker RQR8, led to CAR T-cell depletion. Thus, we here provide the first proof of concept for the generation and preclinical efficacy of CAR T-cells directed against CD117-expressing human hematopoietic cells.

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Fig. 1: CD117 expression in human tissues, healthy human bone marrow, and on AML blast cells.
Fig. 2: Generation and characterization of anti-CD117 CAR T-cells.
Fig. 3: In vitro CAR T-cell activation, proliferation and cytotoxicity depend on-target antigen density.
Fig. 4: Anti-CD117 CAR T-cells deplete healthy human CD117+ CD34+ bone marrow cells and CD117+ AML cells in vitro.
Fig. 5: Anti-CD117 CAR T-cells deplete healthy CD117+ HSPCs in vivo.
Fig. 6: Anti-CD117 CAR T-cells deplete autologous CD117+ AML cells in vivo in humanized mice.
Fig. 7: In vivo depletion of anti-CD117 CAR T-cells with rituximab and ATG.


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This work was supported by research grants of the Swiss National Science Foundation (310030B_166673/1), the Clinical Research Priority Program “Human Hemato-Lymphoid Diseases” of the University of Zurich to MGM; the Clinical Research Priority Program “ImmunoCure” of the University of Zurich to MGM and DN; a Hanne Liebermann-Stiftung grant to AS; a Promedica Foundation grant to RM; a University Research Priority Project Translational Cancer Research grant to RM; a Swiss Cancer Research grant to MGM and DN (KFS-3846-02-2016), as well as a Swiss National Science Foundation grant (No. 310030_182003/1) and “the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program” (grant agreement 670603) to DN.

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RM, JDK, and NFR devised, performed and analyzed experiments, and wrote the paper; NN, DM, and JF performed and analyzed experiments; AS, SP, CFM, and CMW performed experiments; AMM, MvdB, BB, and CM devised experiments and discussed data; and DN and MGM directed the study and wrote the paper.

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Correspondence to Markus G. Manz.

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Myburgh, R., Kiefer, J.D., Russkamp, N.F. et al. Anti-human CD117 CAR T-cells efficiently eliminate healthy and malignant CD117-expressing hematopoietic cells. Leukemia 34, 2688–2703 (2020).

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