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Chemotherapy-resistant human AML stem cells home to and engraft within the bone-marrow endosteal region

Abstract

Acute myelogenous leukemia (AML) is the most common adult leukemia, characterized by the clonal expansion of immature myeloblasts initiating from rare leukemic stem (LS) cells1,2,3. To understand the functional properties of human LS cells, we developed a primary human AML xenotransplantation model using newborn nonobese diabetic/severe combined immunodeficient/interleukin (NOD/SCID/IL)2rγnull mice carrying a complete null mutation of the cytokine γc upon the SCID background4. Using this model, we demonstrated that LS cells exclusively recapitulate AML and retain self-renewal capacity in vivo. They home to and engraft within the osteoblast-rich area of the bone marrow, where AML cells are protected from chemotherapy-induced apoptosis. Quiescence of human LS cells may be a mechanism underlying resistance to cell cycle–dependent cytotoxic therapy. Global transcriptional profiling identified LS cell–specific transcripts that are stable through serial transplantation. These results indicate the potential utility of this AML xenograft model in the development of novel therapeutic strategies targeted at LS cells.

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Figure 1: Self-renewing, long-term engrafting primary human LS cells reside exclusively within the hCD34+hCD38 population.
Figure 2: Primary human LS cells home to and engraft within the bone-marrow osteoblast-rich area, suppressing normal murine hematopoiesis.
Figure 3: Primary hCD34+hCD38 LS cells within the endosteal region exhibit relative resistance to Ara-C induced apoptosis.
Figure 4: Global gene expression profiling of primary human AML and recipient mouse bone-marrow identifies LS cell–specific transcripts.

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Acknowledgements

We thank T. Kanabayashi for the preparation of immunohistochemical staining; N. Aoki for assistance with bone sections; N. Suzuki for technical assistance; N. Kinukawa for assistance with statistical analysis; and F. Ishidate (Carl Zeiss) for assistance with microscopy. This work was supported by the Japan Ministry of Education, Culture, Sports, Science and Technology grant to F.I. and by the US National Institutes of Health grant to L.D.S.

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Contributions

F.I., overall experimental design, transplantation, data analysis, manuscript preparation and discussion; S.Y. transplantation and data analysis; Y.S. overall experimental design, data analysis, statistical analysis, manuscript preparation and discussion; A.H., microarray analysis; H.K., microarray analysis; S.T., flow cytometry; R.N., confocal imaging; T.T., confocal imaging; H.T., flow cytometry; N.S., data analysis; M.F., data analysis; T.M., discussion; B.L., data analysis; K.O., histological analysis; N.U., discussion; S.T., discussion; O.O., microarray analysis and discussion; K.A., discussion; M.H., discussion; L.D.S., discussion.

Corresponding author

Correspondence to Fumihiko Ishikawa.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4 (PDF 4088 kb)

Supplementary Table 1

Serial engraftment of sorted hCD34+hCD38- AML cells. (XLS 53 kb)

Supplementary Table 2

Gene set enrichment analysis identifies genes consistently enriched in hCD34+hCD38 compared with hCD34+hCD38+ cells. (XLS 35 kb)

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Ishikawa, F., Yoshida, S., Saito, Y. et al. Chemotherapy-resistant human AML stem cells home to and engraft within the bone-marrow endosteal region. Nat Biotechnol 25, 1315–1321 (2007). https://doi.org/10.1038/nbt1350

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