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AML-1 is required for megakaryocytic maturation and lymphocytic differentiation, but not for maintenance of hematopoietic stem cells in adult hematopoiesis

Nature Medicine volume 10, pages 299–304 (2004)Cite this article

An Erratum to this article was published on 01 January 2005

Abstract

Embryonic development of multilineage hematopoiesis requires the precisely regulated expression of lineage-specific transcription factors, including AML-1 (encoded by Runx1; also known as CBFA-2 or PEBP-2αB)1,2,3,4,5. In vitro studies and findings in human diseases, including leukemias6,7, myelodysplastic syndromes8 and familial platelet disorder with predisposition to acute myeloid leukemia (AML)9, suggest that AML-1 has a pivotal role in adult hematopoiesis. However, this role has not been fully uncovered in vivo because of the embryonic lethality of Runx1 knockout in mice. Here we assess the requirement of AML-1/Runx1 in adult hematopoiesis using an inducible gene-targeting method10. In the absence of AML-1, hematopoietic progenitors were fully maintained with normal myeloid cell development. However, AML-1-deficient bone marrow showed inhibition of megakaryocytic maturation, increased hematopoietic progenitor cells and defective T- and B-lymphocyte development. AML-1 is thus required for maturation of megakaryocytes and differentiation of T and B cells, but not for maintenance of hematopoietic stem cells (HSCs) in adult hematopoiesis.

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Figure 1: Inducible AML-1 knockout mice.
Figure 2: Thrombocytopenia and megakaryocytic maturation arrest of induced AML-1 knockout mice.
Figure 3: Hematopoietic progenitor cells are expanded in floxed bone marrow.
Figure 4: Competitive repopulation assay.

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References

  1. Okuda, T., van Deursen, J., Hiebert, S.W., Grosveld, G. & Downing, J.R. AML1, the target of multiple chromosomal translocations in human leukemia, is essential for normal fetal liver hematopoiesis. Cell 84, 321–330 (1996).

    Article  CAS  Google Scholar 

  2. Wang, Q. et al. Disruption of the Cbfa2 gene causes necrosis and hemorrhaging in the central nervous system and blocks definitive hematopoiesis. Proc. Natl. Acad. Sci. USA 93, 3444–3449 (1996).

    Article  CAS  Google Scholar 

  3. Mukouyama, Y. et al. The AML1 transcription factor functions to develop and maintain hematogenic precursor cells in the embryonic aorta-gonad-mesonephros region. Dev. Biol. 220, 27–36 (2000).

    Article  CAS  Google Scholar 

  4. Yokomizo, T. et al. Requirement of Runx1/AML1/PEBP2αB for the generation of haematopoietic cells from endothelial cells. Genes Cells 6, 13–23 (2001).

    Article  CAS  Google Scholar 

  5. North, T.E. et al. Runx1 expression marks long-term repopulating hematopoietic stem cells in the midgestation mouse embryo. Immunity 16, 661–672 (2002).

    Article  CAS  Google Scholar 

  6. Tenen, D.G., Hromas, R., Licht, J.D. & Zhang, D.E. Transcription factors, normal myeloid development, and leukemia. Blood 90, 489–519 (1997).

    CAS  Google Scholar 

  7. Lutterbach, B. & Hiebert, S.W. Role of the transcription factor AML-1 in acute leukemia and hematopoietic differentiation. Gene 245, 223–235 (2000).

    Article  CAS  Google Scholar 

  8. Imai, Y. et al. Mutations of the AML1 gene in myelodysplastic syndrome and their functional implications in leukemogenesis. Blood 96, 3154–3160 (2000).

    CAS  Google Scholar 

  9. Song, W.J. et al. Haploinsufficiency of CBFA2 causes familial thrombocytopenia with propensity to develop acute myelogenous leukaemia. Nat. Genet. 23, 166–175 (1999).

    Article  CAS  Google Scholar 

  10. Kuhn, R., Schwenk, F., Aguet, M. & Rajewsky, K. Inducible gene targeting in mice. Science 269, 1427–1429 (1995).

    Article  CAS  Google Scholar 

  11. Jackson, C.W. Cholinesterase as a possible marker for early cells of the megakaryocytic series. Blood 42, 413–421 (1973).

    CAS  Google Scholar 

  12. Kaushansky, K. The enigmatic megakaryocyte gradually reveals its secrets. Bioessays 21, 353–360 (1999).

    Article  CAS  Google Scholar 

  13. Elagib, K.E. et al. RUNX1 and GATA-1 coexpression and cooperation in megakaryocytic differentiation. Blood 101, 4333–4341 (2003).

    Article  CAS  Google Scholar 

  14. Shivdasani, R.A., Fujiwara, Y., McDevitt, M.A. & Orkin, S.H. A lineage-selective knockout establishes the critical role of transcription factor GATA-1 in megakaryocyte growth and platelet development. EMBO J. 16, 3965–3973 (1997).

    Article  CAS  Google Scholar 

  15. Takahashi, S. et al. Role of GATA-1 in proliferation and differentiation of definitive erythroid and megakaryocytic cells in vivo. Blood 92, 434–442 (1998).

    CAS  Google Scholar 

  16. Hodohara, K., Fujii, N., Yamamoto, N. & Kaushansky, K. Stromal cell-derived factor-1 (SDF-1) acts together with thrombopoietin to enhance the development of megakaryocytic progenitor cells (CFU-MK). Blood 95, 769–775 (2000).

    CAS  Google Scholar 

  17. Osawa, M., Hanada, K., Hamada, H. & Nakauchi, H. Long-term lymphohematopoietic reconstitution by a single CD34-low/negative hematopoietic stem cell. Science 273, 242–245 (1996).

    Article  CAS  Google Scholar 

  18. Kondo, M., Weissman, I.L. & Akashi, K. Identification of clonogenic common lymphoid progenitors in mouse bone marrow. Cell 91, 661–672 (1997).

    Article  CAS  Google Scholar 

  19. Rhoades, K.L. et al. Analysis of the role of AML1-ETO in leukemogenesis, using an inducible transgenic mouse model. Blood 96, 2108–2115 (2000).

    CAS  Google Scholar 

  20. Higuchi, M. et al. Expression of a conditional AML1-ETO oncogene bypasses embryonic lethality and establishes a murine model of human t(8;21) acute myeloid leukemia. Cancer Cell 1, 63–74 (2002).

    Article  CAS  Google Scholar 

  21. de Guzman, C.G. et al. Hematopoietic stem cell expansion and distinct myeloid developmental abnormalities in a murine model of the AML1-ETO translocation. Mol. Cell. Biol. 22, 5506–5517 (2002).

    Article  CAS  Google Scholar 

  22. Szilvassy, S.J., Humphries, R.K., Lansdorp, P.M., Eaves, A.C. & Eaves, C.J. Quantitative assay for totipotent reconstituting hematopoietic stem cells by a competitive repopulation strategy. Proc. Natl. Acad. Sci. USA 87, 8736–8740 (1990).

    Article  CAS  Google Scholar 

  23. Lagasse, E. & Weissman, I.L. Flow cytometric identification of murine neutrophils and monocytes. J. Immunol. Methods 197, 139–150 (1996).

    Article  CAS  Google Scholar 

  24. Taniuchi, I. et al. Differential requirements for Runx proteins in CD4 repression and epigenetic silencing during T lymphocyte development. Cell 111, 621–633 (2002).

    Article  CAS  Google Scholar 

  25. Mikkola, H.K. et al. Haematopoietic stem cells retain long-term repopulating activity and multipotency in the absence of stem-cell leukaemia SCL/tal-1 gene. Nature 421, 547–551 (2003).

    Article  CAS  Google Scholar 

  26. Yagi, T. et al. A novel ES cell line, TT2, with high germline-differentiating potency. Anal. Biochem. 214, 70–76 (1993).

    Article  CAS  Google Scholar 

  27. Breton-Gorius, J., Reyes, F., Duhamel, G., Najman, A. & Gorin, N.C. Megakaryoblastic acute leukemia: identification by the ultrastructural demonstration of platelet peroxidase. Blood 51, 45–60 (1978).

    CAS  Google Scholar 

  28. Jackson, C.W., Brown, L.K., Somerville, B.C., Lyles, S.A. & Look, A.T. Two-color flow cytometric measurement of DNA distributions of rat megakaryocytes in unfixed, unfractionated marrow cell suspensions. Blood 63, 768–778 (1984).

    CAS  Google Scholar 

  29. Sherwood, S.W. & Schimke, R.T. Cell cycle analysis of apoptosis using flow cytometry. Methods Cell Biol. 46, 77–97 (1995).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank S. Aizawa for providing us with TT2 ES cells; T. Komori for Runx1 genomic fragments; J. Takeda for Mx-cre-transgenic mice; and thank K. Kumano, A. Kunisato and other members of H.H.'s lab for critical discussion. This work was supported in part by a Grant-in-Aid for Scientific Research (KAKENHI 13307029, 15689015) from the Japan Society for the Promotion of Science, and by Health and Labour Sciences Research grants from the Ministry of Health, Labour and Welfare.

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Author notes

  1. Tetsuya Yamagata

    Present address: Immunology section, Joslin Diabetes Center, Harvard Medical School, One Joslin Place, Boston, Massachusetts, 02215, USA

  2. Hisamaru Hirai: Deceased.

  3. Motoshi Ichikawa and Takashi Asai: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Hematology and Oncology, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan

    Motoshi Ichikawa, Takashi Asai, Toshiki Saito, Go Yamamoto, Sachiko Seo, Tetsuya Yamagata, Shigeru Chiba, Hisamaru Hirai, Seishi Ogawa & Mineo Kurokawa

  2. Department of Regeneration Medicine for Hematopoiesis, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan

    Seishi Ogawa

  3. Department of Clinical Laboratory and Pathology, Inoue Memorial Hospital, 1-16 Shindencho, Chuo-ku, 260-0027, Chiba, Japan

    Ieharu Yamazaki

  4. Department of Hematology, Dokkyo University School of Medicine, 800 Kitakobayashi, Mibu, 321-0293, Tochigi, Japan

    Kinuko Mitani

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Correspondence to Seishi Ogawa.

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Ichikawa, M., Asai, T., Saito, T. et al. AML-1 is required for megakaryocytic maturation and lymphocytic differentiation, but not for maintenance of hematopoietic stem cells in adult hematopoiesis. Nat Med 10, 299–304 (2004). https://doi.org/10.1038/nm997

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