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Transformation from committed progenitor to leukaemia stem cell initiated by MLL–AF9

Abstract

Leukaemias and other cancers possess a rare population of cells capable of the limitless self-renewal necessary for cancer initiation and maintenance1,2,3,4,5,6,7. Eradication of these cancer stem cells is probably a critical part of any successful anti-cancer therapy, and may explain why conventional cancer therapies are often effective in reducing tumour burden, but are only rarely curative. Given that both normal and cancer stem cells are capable of self-renewal, the extent to which cancer stem cells resemble normal tissue stem cells is a critical issue if targeted therapies are to be developed. However, it remains unclear whether cancer stem cells must be phenotypically similar to normal tissue stem cells or whether they can retain the identity of committed progenitors. Here we show that leukaemia stem cells (LSC) can maintain the global identity of the progenitor from which they arose while activating a limited stem-cell- or self-renewal-associated programme. We isolated LSC from leukaemias initiated in committed granulocyte macrophage progenitors through introduction of the MLL–AF9 fusion protein encoded by the t(9;11)(p22;q23). The LSC were capable of transferring leukaemia to secondary recipient mice when only four cells were transferred, and possessed an immunophenotype and global gene expression profile very similar to that of normal granulocyte macrophage progenitors. However, a subset of genes highly expressed in normal haematopoietic stem cells was re-activated in LSC. LSC can thus be generated from committed progenitors without widespread reprogramming of gene expression, and a leukaemia self-renewal-associated signature is activated in the process. Our findings define progression from normal progenitor to cancer stem cell, and suggest that targeting a self-renewal programme expressed in an abnormal context may be possible.

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Figure 1: Leukaemic-GMP are enriched for leukaemia stem cells.
Figure 2: Progenitor-derived leukaemia stem cells maintain progenitor identity and re-activate a self-renewal-associated programme.
Figure 3: The self-renewal-associated signature is found in human MLL -rearranged AML and activated as a hierarchy of gene expression.
Figure 4: Hox genes and Mef2c play a role in L-GMP.

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Acknowledgements

We thank B. Huntly and A. Kung for discussions and ViaCell Inc. for the use of a high-speed cell sorter in the early stages of the project. We thank members of the Akashi laboratory for instruction regarding initial progenitor sorting experiments. We also thank E. Schindler and E. Smith for assistance. This work was funded by the National Institutes of Health, the Leukemia Lymphoma Society, the Richard and Susan Smith foundation, and the Damon Runyon Cancer Research Foundation (to S.A.A.). S.A.A. is a Damon Runyon Clinical Investigator. We dedicate this work to the late Stanley J. Korsmeyer, whose scientific vision inspired the planning of this project.

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Correspondence to Scott A. Armstrong.

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The microarray data discussed here can be found in the Gene Expression Omnibus (GEO) of NCBI at http://www.ncbi.nlm.nih.gov/geo/ through accession numbers GSE3725 and GSE4416 or at http://www.broad.mit.edu/cgi-bin/cancer/publications/pub_menu.cgi. Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Figures 1–10, Supplementary Table 1 and Supplementary Methods

Supplementary Figure 1 details the isolation of normal myeloid progenitor populations and transduction of GMP. Supplementary Figure 2 details the characterization of primary murine leukemias induced by transduction of GMP with an MLL-AF9 retrovirus. Supplementary Figure 3 shows a comparison of colony-forming activity between L-GMP, Lin- Kit-, and lin+ cells. Supplementary Figure 4 details the characterization of leukemias generated by introduction of 20 L-GMP into secondary recipients. Supplementary Figure 5 shows an immunophenotypic analysis of L-GMP propagated in liquid culture (AKLG cells) supplemented with IL3. Supplementary Figure 6 shows the top 50 probe sets for genes that show decreased expression in the HSC and L-GMP as compared to normal progenitors. Supplementary Figure 7 shows the self-renewal associated signature as a subset of the HSC signature: a comparison with a previously published HSC signature. Supplementary Figure 8 shows that The self-renewal associated signature is not merely a prominent portion of the HSC signature. Supplementary Figure 9 shows Mef2c RNA levels assessed by real-time PCR. Supplementary Figure 10 details a model for the development of leukemia stem cells from committed progenitors. Supplementary Table 1 gives details of the mouse transplantation data. (PDF 2175 kb)

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Krivtsov, A., Twomey, D., Feng, Z. et al. Transformation from committed progenitor to leukaemia stem cell initiated by MLL–AF9. Nature 442, 818–822 (2006). https://doi.org/10.1038/nature04980

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