Abstract
Leukemic stem cells (LSCs) are considered a major cause of relapse in acute myeloid leukemia (AML). Defining pathways that control LSC self-renewal is crucial for a better understanding of underlying mechanisms and for the development of targeted therapies. However, currently available culture conditions do not prevent spontaneous differentiation of LSCs, which greatly limits the feasibility of cell-based assays. To overcome these constraints we conducted a high-throughput chemical screen and identified small molecules that inhibit differentiation and support LSC activity in vitro. Similar to reports with cord blood stem cells, several of these compounds suppressed the aryl-hydrocarbon receptor (AhR) pathway, which we show to be inactive in vivo and rapidly activated ex vivo in AML cells. We also identified a compound, UM729, that collaborates with AhR suppressors in preventing AML cell differentiation. Together, these findings provide newly defined culture conditions for improved ex vivo culture of primary human AML cells.
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Acknowledgements
We acknowledge M. Fréchette and A.E. Mejia Alfaro for assistance with xenotransplantation experiments, N. Mayotte and S. Fortier for technical assistance with mouse analyses, J. Duchaine for assistance in screens, and D. Gagné and G. Dulude for technical assistance with high-throughput flow cytometry. We thank specimen donors and the Leukemia Cell Bank of Québec for providing primary AML samples and B. Lehnertz and J. Yeh for critical reading. This work was financially supported by grants from Genome Quebec/Canada to G.S., J.H., S.L. and A.M. and from the Cancer Research Network of the Fonds de recherche du Québec–Santé to J.H. G.S. is supported as a Canada Research Chair in Molecular Genetics of Stem Cells, and J.H. is supported as a Research Chair in Leukemia, supported by Industrielle-Alliance (Université de Montréal). C.P. was supported by a postdoctoral fellowship from the German Cancer Aid (Deutsche Krebshilfe) and is currently supported by a Cole Foundation fellowship.
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C.P. designed, executed and analyzed experiments, wrote the paper and generated the figures. G.S., J.H. and J.K. contributed to project conception, experimental design and editing of the manuscript. I.F. helped identify the compound UM729. S.L. and G.B. contributed to RNA-seq data analysis and statistical analyses. A.M. contributed to chemical data analysis and manuscript editing. R.R. designed and produced compound UM729.
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Pabst, C., Krosl, J., Fares, I. et al. Identification of small molecules that support human leukemia stem cell activity ex vivo. Nat Methods 11, 436–442 (2014). https://doi.org/10.1038/nmeth.2847
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DOI: https://doi.org/10.1038/nmeth.2847
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