Rescue of murine Gata1s mutant M7 leukemic cells by full-length Gata1. Mus musculus

In this project, we studied a mouse model of human Down Syndrome (DS) megakaryocytic leukemia involving mutations in the GATA1 transcription factor (called GATA1s mutation). The model was generated through retroviral insertional mutagenesis in Gata1s mutant fetal liver progenitors. In this study, we analyzed the dependency of these leukemic cells on the Gata1s mutant protein. Here we report Gata1s mutant leukemic cells were dependent on this mutant protein. Introduction of the full-length Gata1 protein to these cells led to their reduced proliferation and increased differentiation along the megakaryocytic lineage. Overall design: We transduced leukemic cells with Gata1/estrogen receptor fusion cDNA (Gata1-ER) and generated stable cell lines. Addition of beta-estradiol to culture medium led to activation of the full-length Gata1 protein in synchronized leukemic cells. Gene expression profiles were collected at multiple time points.

本研究针对携带GATA1转录因子（GATA1 transcription factor）突变（即GATA1s突变）的人类唐氏综合征（Down Syndrome, DS）巨核细胞白血病小鼠模型展开探究。该模型通过在Gata1s突变胎肝祖细胞中进行逆转录病毒插入诱变（retroviral insertional mutagenesis）构建得到。本研究分析了此类白血病细胞对Gata1s突变蛋白的依赖性，结果表明Gata1s突变白血病细胞依赖该突变蛋白。向此类细胞导入全长Gata1蛋白后，其增殖能力降低，且沿巨核细胞谱系的分化水平显著提升。

实验整体设计如下：我们将Gata1/雌激素受体融合互补DNA（cDNA, Gata1-ER）转导至白血病细胞中，构建得到稳定细胞系。向培养基中添加β-雌二醇（beta-estradiol）后，可激活同步化白血病细胞内的全长Gata1蛋白。本研究在多个时间点采集了细胞的基因表达谱。