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Chronic myeloproliferative neoplasms

Azacitidine is effective for targeting leukemia-initiating cells in juvenile myelomonocytic leukemia

Leukemia volume 33, pages 1805–1810 (2019)Cite this article

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Juvenile myelomonocytic leukemia (JMML) is a life-threatening myeloproliferative neoplasm of early childhood [1] originating from multipotent hematopoietic stem/progenitor cells [2] that requires allogeneic hematopoietic stem cell transplantation (HSCT) in the majority of cases [3]. We and others have linked the clinical picture and prognosis of JMML to differential DNA methylation patterns in leukemic cells [4,5,6,7]. A retrospective case series documented that treatment with the DNA methyltransferase-inhibiting agent azacitidine achieved complete clinical and molecular remissions in children diagnosed with JMML, suggesting superior therapeutic potential of this drug [8]. To generate a preclinical research model for JMML, we have previously established a xenotransplantation system of this leukemia in Rag2–/–γc–/– mice [9]. Transplantation of primary JMML cells resulted in stable xenologous engraftment, reproduced a characteristic JMML phenotype, and sustained serial transplantations for up to 1.5 years [9]. Here we used the xenotransplantation system to study if leukemia-initiating cells determine the aberrant DNA methylation profiles in JMML. We then investigated the antileukemic activity of azacitidine in JMML xenografts in comparison to the cytostatic agent, cytosine arabinoside (cytarabine). The latter is used for cytoreduction in JMML but lacks the ability to induce complete remissions [3].

Xenotransplantations were prepared by irradiating neonatal Rag2–/–γc–/– BALB/c mice sublethally with 2.5 Gy. Six hours later, 1 × 106 CD3-depleted JMML mononuclear cells suspended in 30 µl of phosphate-buffered saline were injected intrahepatically. Histopathological examination of experimental animals, isolation of human cells from murine organs, and evaluation of cell populations by flow cytometry were performed as previously described [9]. The experiments were approved by local authorities and complied with the German law for animal protection (Tierversuchsgesetz).

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Acknowledgements

This work was supported by the German Research Foundation (FOR2036 ER599/3-2 to M.E., CRC992-C05 and SPP1463 FL345/4-2 to C.F.).

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    Authors and Affiliations

    1. Division of Pediatric Hematology and Oncology, Department of Pediatrics and Adolescent Medicine, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany

      Christopher F. Krombholz, Lorena Gallego-Villar, Sushree S. Sahoo, Pritam K. Panda, Marcin W. Wlodarski, Charlotte M. Niemeyer, Miriam Erlacher & Christian Flotho

    2. German Cancer Consortium, Heidelberg, Germany

      Marcin W. Wlodarski, Christoph Plass, Charlotte M. Niemeyer, Miriam Erlacher & Christian Flotho

    3. Department of Pathology, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany

      Konrad Aumann

    4. Division of Epigenomics and Cancer Risk Factors, German Cancer Research Center, Heidelberg, Germany

      Mark Hartmann, Daniel B. Lipka, Michael Daskalakis & Christoph Plass

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    1. Christopher F. Krombholz
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    Correspondence to Christian Flotho.

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    These authors contributed equally: Miriam Erlacher, Christian Flotho

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    Krombholz, C.F., Gallego-Villar, L., Sahoo, S.S. et al. Azacitidine is effective for targeting leukemia-initiating cells in juvenile myelomonocytic leukemia. Leukemia 33, 1805–1810 (2019). https://doi.org/10.1038/s41375-018-0343-2

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