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Thrombopoietin-based CAR-T cells demonstrate in vitro and in vivo cytotoxicity to MPL positive acute myelogenous leukemia and hematopoietic stem cells

Gene Therapy volume 29, pages 1–12 (2022)Cite this article

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Abstract

While targeting CD19+ hematologic malignancies with CAR T cell therapy using single chain variable fragments (scFv) has been highly successful, novel strategies for applying CAR T cell therapy with other tumor types are necessary. In the current study, CAR T cells were designed using a ligand binding domain instead of an scFv to target stem-like leukemia cells. Thrombopoietin (TPO), the natural ligand to the myeloproliferative leukemia protein (MPL) receptor, was used as the antigen binding domain to engage MPL expressed on hematopoietic stem cells (HSC) and erythropoietic and megakaryocytic acute myeloid leukemias (AML). TPO-CAR T cells were tested in vitro against AML cell lines with varied MPL expression to test specificity. TPO-CAR T cells were specifically activating and cytotoxic against MPL+ leukemia cell lines. Though the TPO-CAR T cells did not extend survival in vivo, it successfully cleared the MPL+ fraction of leukemia cells. As expected, we also show the TPO-CAR is cytotoxic against MPL expressing bone marrow compartment in AML xenograft models. The data collected demonstrate preclinical potential of TPO-CAR T cells for stem-like leukemia through assessment of targeted killing of MPL+ cells and may facilitate subsequent HSC transplant under reduced intensity conditioning regimens.

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Fig. 1: Establishing MPL as a target in cancer and stem cell biology.
Fig. 2: Generating a thrombopoietin ligand-based CAR to target MPL.
Fig. 3: Cytotoxicity and specificity of the TPO CAR.
Fig. 4: Blocking TPO-CAR activity with thrombopoietin.
Fig. 5: In vivo specificity of TPO-CAR in leukemia xenografts against MPL.

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Acknowledgements

NHLBI 1K08HL141635-01A1 and Atlanta Pediatric Scholars Program K12 Scholar supported by grant K12HD072245.

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Authors and Affiliations

  1. Aflac Cancer and Blood Disorders Center, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA

    Jaquelyn T. Zoine, Chengyu Prince, Jamie Y. Story, Gianna M. Branella, Allison M. Lytle, Andrew Fedanov, Jordan S. Alexander, Christopher C. Porter, Christopher B. Doering, H. Trent Spencer & Shanmuganathan Chandrakasan

  2. Cancer Biology Program, Graduate Division of Biological and Biomedical Sciences, Emory University School of Medicine, Atlanta, GA, USA

    Jaquelyn T. Zoine & Gianna M. Branella

  3. Molecular and Systems Pharmacology Program, Graduate Division of Biological and Biomedical Sciences, Emory University School of Medicine, Atlanta, GA, USA

    Jamie Y. Story & Allison M. Lytle

  4. Children’s Healthcare of Atlanta, Atlanta, GA, USA

    Christopher C. Porter & Shanmuganathan Chandrakasan

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JTZ designed research, performed research, collected data, analyzed/interpreted data, and wrote the manuscript; CZP, JYS, AML, AF, GMB, JYS, and CCP designed research, performed research, collected data, and analyzed/interpreted data. CBD, HTS, and SC designed research, analyzed/interpreted data and edited the manuscript.

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Correspondence to Shanmuganathan Chandrakasan.

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Zoine, J.T., Prince, C., Story, J.Y. et al. Thrombopoietin-based CAR-T cells demonstrate in vitro and in vivo cytotoxicity to MPL positive acute myelogenous leukemia and hematopoietic stem cells. Gene Ther 29, 1–12 (2022). https://doi.org/10.1038/s41434-021-00283-5

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