Skewed megakaryopoiesis in human induced pluripotent stem cell-derived haematopoietic progenitor cells harbouring calreticulin mutations
Skewed megakaryopoiesis in human induced pluripotent stem cell-derived haematopoietic progenitor cells harbouring calreticulin mutations
复制标题
含有钙网蛋白突变的人诱导多能干细胞衍生的造血祖细胞的巨核细胞生成发生偏差
DOI：
10.1111/bjh.15266
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发表时间：
2018
影响因子：
6.5
通讯作者：
Norio Komatsu
中科院分区：
文献类型：
--
作者：
Hiraku Takei;Yoko Edahiro;Shuichi Mano;Nami Masubuchi;Yoshihsa Mizukami;Misa Imai;Soji Morishita;Kyohei Misawa;Tomonori Ochiai;Satoshi Tsuneda;Hiroshi Endo;Sou Nakamura;Koji Eto;Akimichi Ohsaka;Marito Araki;Norio Komatsu
钙网蛋白（CALR）基因的体细胞突变已在大多数JAK 2和MPL未突变的费城染色体阴性骨髓增生性肿瘤（MPN）患者中发现。最近已经表明，突变CALR组成型激活血小板生成素受体MPL，因此，在MPN的发展中起着因果作用。然而，突变CALR在人类造血细胞分化中的作用仍然主要是难以捉摸的。为了检查5碱基插入突变CALR基因（Ins 5）对造血细胞分化的影响，我们从携带aCALR-Ins 5突变的原发性血小板增多症（ET）患者和健康个体（WT）中产生了诱导多能干细胞。巨核细胞生成在Ins 5-造血祖细胞（Ins 5-HPC）中比在WT-HPC中更突出，这意味着该系统重现了在CALR-突变ET患者骨髓中观察到的巨核细胞增多。Ins 5-HPC表现出GATA 1和GATA 2的表达水平升高，表明祖细胞过早地向巨核细胞分化。我们还证明了3-羟基阿那格雷显著干扰巨核细胞生成，但不干扰红细胞生成。综上所述，我们建立了一个体外模型系统，重现了由MUCALR引起的巨核细胞生成。该系统可用于验证携带CALR突变的MPN患者的治疗化合物，以及对人类血液细胞分化中突变CALR的详细研究。
Somatic mutations in thecalreticulin(CALR) gene have been found in most patients withJAK2‐ andMPL‐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 mutantCALRgene (Ins5) on haematopoietic cell differentiation, we generated induced pluripotent stem cells from an essential thrombocythaemia (ET) patient harbouring aCALR‐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 ofCALR‐mutant ET patients. Ins5‐HPCs exhibited elevated expression levels ofGATA1andGATA2,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 anin vitromodel system that recapitulates megakaryopoiesis caused by mutantCALR. This system can be used to validate therapeutic compounds for MPN patients harbouringCALRmutations and in detailed studies on mutant CALR in human haematological cell differentiation.