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Immunotherapy

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

Leukemia volume 34, pages 2688–2703 (2020)Cite this article

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Abstract

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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Acknowledgements

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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  1. These authors contributed equally: Renier Myburgh, Jonathan D. Kiefer, Norman F. Russkamp

Authors and Affiliations

  1. Department of Medical Oncology and Hematology, University Hospital Zurich and University of Zurich, Comprehensive Cancer Center Zurich (CCCZ), Zurich, Switzerland

    Renier Myburgh, Jonathan D. Kiefer, Norman F. Russkamp, Chiara F. Magnani, Alexander Simonis, Surema Pfister, C. Matthias Wilk, Antonia M. Müller & Markus G. Manz

  2. Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zurich, Comprehensive Cancer Center Zurich (CCCZ), Zurich, Switzerland

    Jonathan D. Kiefer & Dario Neri

  3. Institute of Experimental Immunology, University of Zurich, Comprehensive Cancer Center Zurich (CCCZ), Zurich, Switzerland

    Nicolás Nuñez, Donal McHugh, Burkhard Becher, Christian Münz & Maries van den Broek

  4. Institute of Pathology and Molecular Pathology, University and University Hospital Zurich, Comprehensive Cancer Center Zurich (CCCZ), Zurich, Switzerland

    Juliane Friemel

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  1. Renier Myburgh
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Contributions

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). https://doi.org/10.1038/s41375-020-0818-9

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