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Impaired hematopoietic differentiation of RUNX1-mutated induced pluripotent stem cells derived from FPD/AML patients

Leukemia volume 28, pages 2344–2354 (2014)Cite this article

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Abstract

Somatic mutation of RUNX1 is implicated in various hematological malignancies, including myelodysplastic syndrome and acute myeloid leukemia (AML), and previous studies using mouse models disclosed its critical roles in hematopoiesis. However, the role of RUNX1 in human hematopoiesis has never been tested in experimental settings. Familial platelet disorder (FPD)/AML is an autosomal dominant disorder caused by germline mutation of RUNX1, marked by thrombocytopenia and propensity to acute leukemia. To investigate the physiological function of RUNX1 in human hematopoiesis and pathophysiology of FPD/AML, we derived induced pluripotent stem cells (iPSCs) from three distinct FPD/AML pedigrees (FPD-iPSCs) and examined their defects in hematopoietic differentiation. By in vitro differentiation assays, FPD-iPSCs were clearly defective in the emergence of hematopoietic progenitors and differentiation of megakaryocytes, and overexpression of wild-type (WT)-RUNX1 reversed most of these phenotypes. We further demonstrated that overexpression of mutant-RUNX1 in WT-iPSCs did not recapitulate the phenotype of FPD-iPSCs, showing that the mutations were of loss-of-function type. Taken together, this study demonstrated that haploinsufficient RUNX1 allele imposed cell-intrinsic defects on hematopoietic differentiation in human experimental settings and revealed differential impacts of RUNX1 dosage on human and murine megakaryopoiesis. FPD-iPSCs will be a useful tool to investigate mutant RUNX1-mediated molecular processes in hematopoiesis and leukemogenesis.

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Acknowledgements

We thank excellent technical assistance by J Kawakita. We also thank S Suzuki (FACS Core Laboratory, Keio University School of Medicine) for FACS sorting, W Akamatsu and M Sato (Department of Physiology, Keio University School of Medicine) and H Nakata (Division of Cardiology, Keio University School of Medicine) for cell culture or establishment of iPSCs. This work was supported in part by a grant from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Author notes

  1. Y Harada & H Harada

    Present address: 12Present address: Department of Hematology, Juntendo University School of Medicine, Tokyo, Japan.,

Authors and Affiliations

  1. Division of Hematology, Department of Internal Medicine, Keio University School of Medicine, Tokyo, Japan

    M Sakurai, H Kunimoto, Y Fukuchi, K Sadahira, S Okamoto & H Nakajima

  2. Department of Transfusion Medicine and Cell Therapy, Keio University School of Medicine, Tokyo, Japan

    N Watanabe

  3. Division of Cardiology, Department of Internal Medicine, Keio University School of Medicine, Tokyo, Japan

    S Yuasa & K Fukuda

  4. Japan Science and Technology Agency, ERATO, Tokyo, Japan

    S Yamazaki & H Nakauchi

  5. Division of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    T Nishimura & H Nakauchi

  6. Department of Physiology, Keio University School of Medicine, Tokyo, Japan

    H Okano

  7. Division of Hematology and Rheumatology, Department of Internal Medicine, Kinki University Faculty of Medicine, Osaka, Japan

    Y Morita & I Matsumura

  8. Department of Pediatrics, Hirosaki University Graduate School of Medicine, Aomori, Japan

    K Kudo & E Ito

  9. Department of Pediatric Hematology/Oncology, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    Y Ebihara & K Tsuji

  10. Division of Stem Cell Processing, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    K Tsuji

  11. Department of Hematology and Oncology, Division of Clinical Research, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan

    Y Harada & H Harada

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Correspondence to H Nakajima.

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Competing interests

HO is the scientific consultant of San Bio, Inc., Eisai Co Ltd. and Daiichi Sankyo Co Ltd. HN is a founder, a member of scientific advisory board and shareholder of ReproCELL and is a scientific advisor of Megakaryon, iCELL and Shionogi & Co. The remaining authors declare no conflict of interest.

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Sakurai, M., Kunimoto, H., Watanabe, N. et al. Impaired hematopoietic differentiation of RUNX1-mutated induced pluripotent stem cells derived from FPD/AML patients. Leukemia 28, 2344–2354 (2014). https://doi.org/10.1038/leu.2014.136

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