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Oncogenes, Fusion Genes and Tumor Suppressor Genes

Ligand-independent FLT3 activation does not cooperate with MLL-AF4 to immortalize/transform cord blood CD34+ cells

Leukemia volume 28, pages 666–674 (2014)Cite this article

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Abstract

MLL-AF4 fusion is hallmark in high-risk infant pro-B-acute lymphoblastic leukemia (pro-B-ALL). Our limited understanding of MLL-AF4-mediated transformation reflects the absence of human models reproducing this leukemia. Hematopoietic stem/progenitor cells (HSPCs) constitute likely targets for transformation. We previously reported that MLL-AF4 enhanced hematopoietic engraftment and clonogenic potential in cord blood (CB)-derived CD34+ HSPCs but was not sufficient for leukemogenesis, suggesting that additional oncogenic lesions are required for MLL-AF4-mediated transformation. MLL-AF4+ pro-B-ALL display enormous levels of FLT3, and occasionally FLT3-activating mutations, thus representing a candidate cooperating event in MLL-AF4+ pro-B-ALL. We have explored whether FLT3.TKD (tyrosine kinase domain) mutation or increased expression of FLT3.WT (wild type) cooperates with MLL-AF4 to immortalize/transform CB-CD34+ HSPCs. In vivo, FLT3.TKD/FLT3.WT alone, or in combination with MLL-AF4, enhances hematopoietic repopulating function of CB-CD34+ HSPCs without impairing migration or hematopoietic differentiation. None of the animals transplanted with MLL-AF4+FLT3.TKD/WT-CD34+ HSPCs showed any sign of disease after 16 weeks. In vitro, enforced expression of FLT3.TKD/FLT3.WT conveys a transient overexpansion of MLL-AF4-expressing CD34+ HSPCs associated to higher proportion of cycling cells coupled to lower apoptotic levels, but does not augment clonogenic potential nor confer stable replating. Together, FLT3 activation does not suffice to immortalize/transform MLL-AF4-expressing CB-CD34+ HSPCs, suggesting the need of alternative (epi)-genetic cooperating oncogenic lesions.

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Acknowledgements

This work was funded by FIS/FEDER (PI10/00449 to PM and PI11/00119 to CB), The Spanish Association Against Cancer to PM. CB is supported by a ‘Miguel Servet’ Fellowship (CP07/0059). RMo is supported by The ISCIII (CA10/01332). DR-M and CPri are supported by PFIS scholarships. PM is an ICREA investigator. His research is supported by ISCIII Red de Terapia Celular (Tercel; RD12/0019/0006).

Author contributions

RMo, VA, CB and PM designed the research; RMo, VA, CB, AB, RMa, CC, PJR, DR-M, OM-N and CP performed the research and analyzed the data; and RMo, CB and PM wrote the paper. The manuscript has been seen and approved by all authors.

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Authors and Affiliations

  1. GENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government, Granada, Spain

    R Montes, V Ayllón, C Prieto, D Romero-Moya, P J Real, O Navarro-Montero, C Bueno & P Menendez

  2. Department of Stem Cells, Faculty of Medicine, Development and Cancer, Cell Therapy Program of the University of Barcelona, Josep Carreras Leukemia Research Institute, Barcelona, Spain

    C Prieto, D Romero-Moya, C Bueno & P Menendez

  3. Institute of Pharmaceutical Biology/ZAFES/DCAL, Goethe-University of Frankfurt, Biocenter, Frankfurt, Germany

    A Bursen, C Prelle & R Marschalek

  4. Hospital Universitario de Salamanca, Servicio de Hematología, Salamanca, Spain

    C Chillón

  5. Instituciò Catalana de Reserca i Estudis Avançats (ICREA),

    P Menendez

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  1. R Montes
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Correspondence to C Bueno or P Menendez.

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Montes, R., Ayllón, V., Prieto, C. et al. Ligand-independent FLT3 activation does not cooperate with MLL-AF4 to immortalize/transform cord blood CD34+ cells. Leukemia 28, 666–674 (2014). https://doi.org/10.1038/leu.2013.346

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