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Animal models

Unexpected variation in leukemia stem cell frequency and genetic heterogeneity in two murine leukemia models initiated by AML1/ETO9a and CALM/AF10

Leukemia volume 34, pages 1706–1710 (2020)Cite this article

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Acute myeloid leukemia (AML) is an extremely aggressive, genetically heterogeneous disorder requiring the acquisition of multiple (two to eight) genetic aberrations which act in concert and result in the malignant transformation of a hematopoietic cell [1]. Advances in molecular techniques, particularly the advent of next generation sequencing, has led to the identification of a plethora of genetic lesions and greatly increased our understanding of the molecular heterogeneity of AML [2]. However, one of the major challenges is to determine which of the identified mutations contribute to the disease process as so-called driver mutation. Further, to add to the complex genomic landscape of AML, it is now well known that heterogeneity is found not only between patients (intertumor heterogeneity) but also within individual leukemias (intratumor heterogeneity) [3]. The leukemia stem cell model provides an explanation for the functional and phenotypic heterogeneity observed within a tumor. The model posits that within the leukemic bulk, only a small subset of cells, the leukemia stem cells (LSC) possess the ability to propagate the leukemia phenotype, and are largely responsible for therapy failure and relapse in leukemia patients [4, 5]. Although a number of animal models of AML have been established, there remains a lack of clinically relevant models that recapitulate this intra and intertumor heterogeneity observed in human AML.

In this study, we established two retroviral transduction, murine bone marrow transplantation leukemia models (MBMTM), in order to mimic the genetic, functional, and phenotypic heterogeneity of AML. The first model used the AML1/ETO9a (A/E9a) fusion, an alternatively spliced isoform of AML1/ETO (A/E), also known as RUNX-RUNX1T1, as a basal driver oncogene (Fig. S1a) [6]. The second model used the CALM/AF10-minimal fusion (C/A-MF), which contains the leukemogenic regions of CALM/AF10 (C/A) also known as PICALM/MLLT10 (Fig. S1b), a rare but recurring fusion gene observed in AML, acute lymphoblastic leukemia, and malignant lymphoma [7,8,9].

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Fig. 1: Identifying potential cooperating mutations in A/E9a and C/A-MF murine leukemias.
Fig. 2: The murine leukemias recapitulate the GEP of patient samples and show varied leukemia stem cell frequencies.

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Acknowledgements

This work is supported by Leukemia and Blood Cancer New Zealand, the family of Marijana Kumerich and Cancer Research Trust New Zealand.

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  1. These authors contributed equally: Rhea Desai, Sarvenaz Taghavi

Authors and Affiliations

  1. Leukaemia & Blood Cancer Research Unit, Department of Molecular Medicine and Pathology, Faculty of Medical and Health Sciences, The University of Auckland, Auckland, New Zealand

    Rhea Desai, Jessica C. Chase, Martin Chopra, Jenny Chien, Peter J. Browett, Purvi M. Kakadia & Stefan K. Bohlander

  2. Melbourne Brain Centre, Flow Cytometry Facility, The University of Melbourne, Melbourne, VIC, Australia

    Sarvenaz Taghavi

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  1. Rhea Desai
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Contributions

RD performed the experiments, analyzed the data, and wrote the paper. ST performed the experiments and analyzed the data. JCC analyzed the RNA sequencing data. MC helped in the design and set up of the in vivo experiments. JC provided technical help with sequencing of the whole exome and transcriptome libraries. PJB provided helpful discussion during the course of the study. PMK designed the experiments, sequenced the transcriptome, and whole exome samples, analyzed the WES sequencing data, and cosupervised the study. SKB designed the experiments, supervised the study, analyzed the data, reviewed, and approved the paper.

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Correspondence to Stefan K. Bohlander.

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Desai, R., Taghavi, S., Chase, J.C. et al. Unexpected variation in leukemia stem cell frequency and genetic heterogeneity in two murine leukemia models initiated by AML1/ETO9a and CALM/AF10. Leukemia 34, 1706–1710 (2020). https://doi.org/10.1038/s41375-019-0657-8

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