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Induction of cell cycle entry eliminates human leukemia stem cells in a mouse model of AML

Nature Biotechnology volume 28, pages 275–280 (2010)Cite this article

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Abstract

Cancer stem cells have been proposed to be important for initiation, maintenance and recurrence of various malignancies, including acute myeloid leukemia (AML)1,2,3. We have previously reported4 that CD34+CD38− human primary AML stem cells residing in the endosteal region of the bone marrow are relatively chemotherapy resistant. Using a NOD/SCID/IL2rγnull mouse model of human AML, we now show that the AML stem cells in the endosteal region are cell cycle quiescent and that these stem cells can be induced to enter the cell cycle by treatment with granulocyte colony-stimulating factor (G-CSF). In combination with cell cycle-dependent chemotherapy, G-CSF treatment significantly enhances induction of apoptosis and elimination of human primary AML stem cells in vivo. The combination therapy leads to significantly increased survival of secondary recipients after transplantation of leukemia cells compared with chemotherapy alone.

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Figure 1: Quiescent LSCs enter cell cycle after in vivo G-CSF treatment.
Figure 2: Quiescent human AML cells within the bone marrow endosteal region enter the cell cycle after in vivo G-CSF treatment.
Figure 3: Cell cycle entry potentiates chemotherapy-induced apoptosis of LSCs in vivo, reduces LSC frequency and leads to superior survival of secondary recipients.

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Acknowledgements

We thank T. Tanaka and M. Yamaguchi for assistance with confocal microscopy. We thank T. Kanabayashi for assistance with histological studies. We thank M. Narita for assistance with patient data collection. This work was supported through grants from the Ministry of Education, Culture, Sports, Science and Technology-Japan, National Institute of Biomedical Innovation, Takeda Science Foundation and Uehara Memorial Foundation to F.I. and National Institutes of Health grant CA20408 to L.D.S.

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Authors and Affiliations

  1. Research Unit for Human Disease Models, RIKEN Research Center for Allergy and Immunology, Yokohama, Japan

    Yoriko Saito, Nahoko Suzuki, Mariko Tomizawa-Murasawa, Akiko Sone, Yuho Najima, Shinsuke Takagi, Yuki Aoki & Fumihiko Ishikawa

  2. Department of Hematology, Toranomon Hospital, Tokyo, Japan

    Naoyuki Uchida, Shinsuke Takagi, Atsushi Wake & Shuichi Taniguchi

  3. Nippon Becton Dickinson Company, Tokyo, Japan

    Satoshi Tanaka

  4. The Jackson Laboratory, Bar Harbor, Maine, USA

    Leonard D Shultz

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  1. Yoriko Saito
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Contributions

Y.S., L.D.S. and F.I. designed the study, analyzed the data and wrote the manuscript. N.U., A.W. and S. Taniguchi provided clinical samples, information and discussion. Y.S., S. Tanaka, M.T.-M., N.S., A.S. and F.I. performed the experiments. S. Takagi and Y.A. analyzed the data. Y.N. analyzed the data and wrote the manuscript.

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Correspondence to Fumihiko Ishikawa.

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Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–5 and Supplementary Tables 1–5 (PDF 9230 kb)

Supplementary Movie 1

This movie shows a cross-sectional view through a three-dimensional reconstruction of the recipient femur shown in Fig. 2j. The bone section was labeled for human CD45 (red), Ki67 (green) and DAPI (blue). The serial cross-sectional images along the long axis of the bone demonstrate that there is little to no Ki67 expression by hCD45+DAPI+ AML cells in the BM endosteal region adjacent to the bone at baseline. (MOV 8927 kb)

Supplementary Movie 2

This movie shows a cross-sectional view through a three-dimensional reconstruction of the recipient femur shown in Fig. 2k. The bone section was labeled for human CD45 (red), Ki67 (green) and DAPI (blue). The serial cross-sectional images along the long axis of the bone demonstrate the appearance of Ki67+hCD45+DAPI+ cells within the BM endosteal region following G-CSF treatment. (MOV 7826 kb)

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Saito, Y., Uchida, N., Tanaka, S. et al. Induction of cell cycle entry eliminates human leukemia stem cells in a mouse model of AML. Nat Biotechnol 28, 275–280 (2010). https://doi.org/10.1038/nbt.1607

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