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Skewed megakaryopoiesis in human induced pluripotent stem cell‐derived haematopoietic progenitor cells harbouring calreticulin mutations
Author(s) -
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
Publication year - 2018
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/bjh.15266
Subject(s) - haematopoiesis , biology , stem cell , calreticulin , progenitor cell , cancer research , induced pluripotent stem cell , cellular differentiation , microbiology and biotechnology , genetics , gene , embryonic stem cell , endoplasmic reticulum
Summary Somatic mutations in the calreticulin ( CALR ) gene have been found in most patients with JAK 2 ‐ and MPL ‐unmutated Philadelphia chromosome‐negative myeloproliferative neoplasms ( MPN s). It has recently been shown that mutant CALR constitutively activates the thrombopoietin receptor MPL and, thus, plays a causal role in the development of MPN s. 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‐ HPC s) than in WT ‐ HPC s, implying that the system recapitulates megakaryocytosis observed in the bone marrow of CALR ‐mutant ET patients. Ins5‐ HPC s exhibited elevated expression levels of GATA 1 and GATA 2, 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.