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Molecular targets for therapy

Modeling human MLL-AF9 translocated acute myeloid leukemia from single donors reveals RET as a potential therapeutic target

Leukemia volume 31, pages 1166–1176 (2017)Cite this article

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Abstract

Acute myeloid leukemias (AMLs) result from a series of genetic events occurring in a stem or progenitor hematopoietic cell that gives rise to their clonal expansion and an impaired capacity to differentiate. To circumvent the genetic heterogeneity of AML patient cohorts, we have developed a model system, driven by the MLL-AF9 (MA9) oncogene, to generate multiple human leukemias using progenitor cells from a single healthy donor. Through stepwise RNA-sequencing data generated using this model and AML patients, we have identified consistent changes associated with MA9-driven leukemogenesis and demonstrate that no recurrent secondary mutations are required. We identify 39 biomarkers whose high expression level is specific to this genetic subtype of AML and validate that many of these have diagnostic utility. We further examined one biomarker, the receptor tyrosine kinase (RTK) RET, and show through shRNA knockdowns that its expression is essential for in vivo and in vitro growth of MA9-AML. These results highlight the value of novel human models of AML derived from single donors using specific oncogenic fusions to understand their biology and to uncover potential therapeutic targets.

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Acknowledgements

This work was supported by grants from the Cole Foundation (to BTW, JH, MC—fellowship), the Fonds de Recherche du Québec en Santé (to BTW), Cancer Research Society (to BTW) and the Leukemia & Lymphoma Society of Canada (to FB) and CIHR (to FB). Human leukemia specimens were collected and analyzed by the Banque de cellules leucémiques du Québec (BCLQ), supported by the Cancer Research Network of the Fonds de Recherche du Québec en Santé (FRQS). We would like to thank the clinicians and nurses at CHU de Québec and Hotel Dieu de Lévis for cord blood collection and we also wish to acknowledge the contribution of all of the courageous patients who provided samples used in this study. We would like to acknowledge Josette-Renee Landry for critical comments on the manuscript and Marianne Arteau, Jennifer Huber, Raphaëlle Lambert and Danièle Gagné for expert technical assistance with experimental work.

Author contributions

BTW and FB designed the study, analyzed data and wrote the manuscript with help from LG, MC and AB. LG and AB engineered human leukemias in vivo from cord blood CD34+ cells. VL, ÉR, KL and AF were responsible for molecular biology work, cloning and cell culture work and RJ performed FACS analysis. ABP and EG generated the dual shRNA vector and tested these in cell lines with help from JP. JH and SC were responsible for banking of pediatric AML samples and JH analyzed cytogenetic studies of the human AML cells while JS, LP and SC isolated normal blood cell populations for RNA-seq. All authors discussed the results and participated in the revision of the manuscript.

Data deposition

All sequencing data has been deposited to GEO and is accessible through the following accession numbers: GSE71686, GSE71689, GSE70755, GSE71800, GSE71840, GSE72041 and GSE72061 within the GEO SuperSeries GSE71691. All whole-exome sequencing raw data were submitted to SRA database (SRA (http://trace.ncbi.nlm.nih.gov/Traces/sra/) accession number SRP062510).

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Authors and Affiliations

  1. Centre de recherche en infectiologie du CHUL, Centre de recherche du CHU de Québec, Quebec City, Quebec, Canada

    F Barabé, L Gil, A Bergeron & J Simard

  2. CHU de Québec Hôpital Enfant-Jésus, Quebec City, Quebec, Canada

    F Barabé

  3. Department of Medicine, Université Laval, Quebec City, Quebec, Canada

    F Barabé

  4. Laboratory for High throughput biology, Ste-Justine Hospital, Université de Montréal, Montréal, Quebec, Canada

    M Celton, V Lamontagne, É Roques, K Lagacé, A Forest, R Johnson & B T Wilhelm

  5. Institute for Research in Immunology and Cancer, Ste-Justine Hospital, Université de Montréal, Montréal, Quebec, Canada

    M Celton, V Lamontagne, É Roques, K Lagacé, A Forest, R Johnson, J Pelloux, A Bellemare-Pelletier, E Gagnon & B T Wilhelm

  6. Department of Pediatrics, Division of Hematology, Ste-Justine Hospital, Université de Montréal, Montréal, Quebec, Canada

    L Pécheux & S Cellot

  7. Laboratory for Cancer Immunobiology, Maisonneuve-Rosemont Hospital, Montréal, Quebec, Canada

    J Pelloux, A Bellemare-Pelletier & E Gagnon

  8. Department of Microbiology and Immunology, Maisonneuve-Rosemont Hospital, Montréal, Quebec, Canada

    E Gagnon

  9. Division of Hematology-Oncology and Leukemia Cell Bank of Quebec, Maisonneuve-Rosemont Hospital, Montréal, Quebec, Canada

    J Hébert

  10. Department of Medicine, Université de Montréal, Montréal, Quebec, Canada

    J Hébert & B T Wilhelm

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  1. F Barabé
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  2. L Gil
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Correspondence to F Barabé or B T Wilhelm.

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Barabé, F., Gil, L., Celton, M. et al. Modeling human MLL-AF9 translocated acute myeloid leukemia from single donors reveals RET as a potential therapeutic target. Leukemia 31, 1166–1176 (2017). https://doi.org/10.1038/leu.2016.302

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