Skewed megakaryopoiesis in human induced pluripotent stem cell-derived haematopoietic progenitor cells harbouring calreticulin mutations

Hiraku Takei, Yoko Edahiro, Shuichi Mano, Nami Masubuchi, Yoshihisa Mizukami, Misa Imai, Soji Morishita, Kyohei Misawa, Tomonori Ochiai, Satoshi Tsuneda, Hiroshi Endo, Sou Nakamura, Koji Eto, Akimichi Ohsaka, Marito Araki, Norio Komatsu

    Research output: Contribution to journalArticle

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    Abstract

    Somatic mutations in the calreticulin (CALR) gene have been found in most patients with JAK2- and MPL-unmutated Philadelphia chromosome-negative myeloproliferative neoplasms (MPNs). It has recently been shown that mutant CALR constitutively activates the thrombopoietin receptor MPL and, thus, plays a causal role in the development of MPNs. However, the roles of mutant CALR in human haematopoietic cell differentiation remain predominantly elusive. To examine the impact of the 5-base insertion mutant CALR gene (Ins5) on haematopoietic cell differentiation, we generated induced pluripotent stem cells from an essential thrombocythaemia (ET) patient harbouring a CALR-Ins5 mutation and from a healthy individual (WT). Megakaryopoiesis was more prominent in Ins5-haematopoietic progenitor cells (Ins5-HPCs) than in WT-HPCs, implying that the system recapitulates megakaryocytosis observed in the bone marrow of CALR-mutant ET patients. Ins5-HPCs exhibited elevated expression levels of GATA1 and GATA2, suggesting a premature commitment to megakaryocytic differentiation in progenitor cells. We also demonstrated that 3-hydroxy anagrelide markedly perturbed megakaryopoiesis, but not erythropoiesis. Collectively, we established an in vitro model system that recapitulates megakaryopoiesis caused by mutant CALR. This system can be used to validate therapeutic compounds for MPN patients harbouring CALR mutations and in detailed studies on mutant CALR in human haematological cell differentiation.

    Original languageEnglish
    JournalBritish Journal of Haematology
    DOIs
    Publication statusAccepted/In press - 2018 Jan 1

    Fingerprint

    Calreticulin
    Induced Pluripotent Stem Cells
    Hematopoietic Stem Cells
    Mutation
    Essential Thrombocythemia
    Cell Differentiation
    Thrombopoietin Receptors
    Philadelphia Chromosome
    Neoplasms
    Erythropoiesis
    Genes
    Stem Cells
    Bone Marrow

    Keywords

    • Anagrelide
    • Calreticulin
    • Induced pluripotent stem cells
    • Megakaryopoiesis
    • Myeloproliferative neoplasm

    ASJC Scopus subject areas

    • Hematology

    Cite this

    Skewed megakaryopoiesis in human induced pluripotent stem cell-derived haematopoietic progenitor cells harbouring calreticulin mutations. / Takei, Hiraku; Edahiro, Yoko; Mano, Shuichi; Masubuchi, Nami; Mizukami, Yoshihisa; Imai, Misa; Morishita, Soji; Misawa, Kyohei; Ochiai, Tomonori; Tsuneda, Satoshi; Endo, Hiroshi; Nakamura, Sou; Eto, Koji; Ohsaka, Akimichi; Araki, Marito; Komatsu, Norio.

    In: British Journal of Haematology, 01.01.2018.

    Research output: Contribution to journalArticle

    Takei, H, Edahiro, Y, Mano, S, Masubuchi, N, Mizukami, Y, Imai, M, Morishita, S, Misawa, K, Ochiai, T, Tsuneda, S, Endo, H, Nakamura, S, Eto, K, Ohsaka, A, Araki, M & Komatsu, N 2018, 'Skewed megakaryopoiesis in human induced pluripotent stem cell-derived haematopoietic progenitor cells harbouring calreticulin mutations' British Journal of Haematology. https://doi.org/10.1111/bjh.15266
    Takei, Hiraku ; Edahiro, Yoko ; Mano, Shuichi ; Masubuchi, Nami ; Mizukami, Yoshihisa ; Imai, Misa ; Morishita, Soji ; Misawa, Kyohei ; Ochiai, Tomonori ; Tsuneda, Satoshi ; Endo, Hiroshi ; Nakamura, Sou ; Eto, Koji ; Ohsaka, Akimichi ; Araki, Marito ; Komatsu, Norio. / Skewed megakaryopoiesis in human induced pluripotent stem cell-derived haematopoietic progenitor cells harbouring calreticulin mutations. In: British Journal of Haematology. 2018.
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    abstract = "Somatic mutations in the calreticulin (CALR) gene have been found in most patients with JAK2- and MPL-unmutated Philadelphia chromosome-negative myeloproliferative neoplasms (MPNs). It has recently been shown that mutant CALR constitutively activates the thrombopoietin receptor MPL and, thus, plays a causal role in the development of MPNs. However, the roles of mutant CALR in human haematopoietic cell differentiation remain predominantly elusive. To examine the impact of the 5-base insertion mutant CALR gene (Ins5) on haematopoietic cell differentiation, we generated induced pluripotent stem cells from an essential thrombocythaemia (ET) patient harbouring a CALR-Ins5 mutation and from a healthy individual (WT). Megakaryopoiesis was more prominent in Ins5-haematopoietic progenitor cells (Ins5-HPCs) than in WT-HPCs, implying that the system recapitulates megakaryocytosis observed in the bone marrow of CALR-mutant ET patients. Ins5-HPCs exhibited elevated expression levels of GATA1 and GATA2, suggesting a premature commitment to megakaryocytic differentiation in progenitor cells. We also demonstrated that 3-hydroxy anagrelide markedly perturbed megakaryopoiesis, but not erythropoiesis. Collectively, we established an in vitro model system that recapitulates megakaryopoiesis caused by mutant CALR. This system can be used to validate therapeutic compounds for MPN patients harbouring CALR mutations and in detailed studies on mutant CALR in human haematological cell differentiation.",
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    AU - Edahiro, Yoko

    AU - Mano, Shuichi

    AU - Masubuchi, Nami

    AU - Mizukami, Yoshihisa

    AU - Imai, Misa

    AU - Morishita, Soji

    AU - Misawa, Kyohei

    AU - Ochiai, Tomonori

    AU - Tsuneda, Satoshi

    AU - Endo, Hiroshi

    AU - Nakamura, Sou

    AU - Eto, Koji

    AU - Ohsaka, Akimichi

    AU - Araki, Marito

    AU - Komatsu, Norio

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