Impaired hematopoietic differentiation of RUNX1-mutated induced pluripotent stem cells derived from FPD/AML patients
Impaired hematopoietic differentiation of RUNX1-mutated induced pluripotent stem cells derived from FPD/AML patients
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DOI：
10.1038/leu.2014.136
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发表时间：
2014-12-01
期刊：
影响因子：
11.4
通讯作者：
Nakajima, H.
中科院分区：
文献类型：
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作者：
Sakurai, M.;Kunimoto, H.;Nakajima, H.
RUNX1的体细胞突变与多种血液恶性肿瘤有关，包括骨髓增生异常综合征和急性髓系白血病（AML），之前使用小鼠模型的研究揭示了其在造血中的关键作用。然而，RUNX1 在人类造血中的作用从未在实验环境中得到测试。家族性血小板疾病 (FPD)/AML 是一种由 RUNX1 种系突变引起的常染色体显性遗传疾病，其特点是血小板减少和急性白血病倾向。为了研究 RUNX1 在人类造血和 FPD/AML 病理生理学中的生理功能，我们从三个不同的 FPD/AML 谱系 (FPD-iPSC) 中衍生出诱导多能干细胞 (iPSC)，并检查它们在造血分化方面的缺陷。通过体外分化测定，FPD-iPSC 在造血祖细胞的出现和巨核细胞的分化方面存在明显缺陷，并且野生型 (WT)-RUNX1 的过度表达逆转了大多数这些表型。我们进一步证明，WT-iPSC 中突变体 RUNX1 的过表达并不能重现 FPD-iPSC 的表型，表明该突变属于功能丧失型。总而言之，这项研究证明，单倍体不足的 RUNX1 等位基因在人类实验环境中对造血分化造成了细胞内在缺陷，并揭示了 RUNX1 剂量对人类和小鼠巨核细胞生成的不同影响。 FPD-iPSC 将成为研究突变 RUNX1 介导的造血和白血病发生分子过程的有用工具。
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.