University of Rochester Medical Center, Department of Pediatrics, Center for Pediatric Biomedical Research, Rochester, NY, USA.
Laboratoire de Génétique et Biophysique des Plantes, CNRS-CEA-Aix Marseille Université, Marseille, France.
Exp Hematol. 2014 Jul;42(7):536-46.e8. doi: 10.1016/j.exphem.2014.03.006. Epub 2014 Apr 2.
Globin gene regulation occurs in the context of a maturing erythroid cell, which is undergoing significant changes in chromatin structure and gene expression. There are few model systems available that facilitate studies of globin gene regulation in the context of erythroid maturation. Extensively self-renewing erythroblasts (ESREs) are a nontransformed model of erythroid maturation derived from murine fetal liver or yolk sac. Imaging flow cytometry and RNA-seq studies demonstrate that ESREs functionally and molecularly model erythroid maturation. To address the need for a model system that also recapitulates human globin switching, ESREs were derived from mice transgenic for the complete human β-globin locus (β-yac ESREs). β-yac ESREs express β-globin from the transgenic human locus, with minimal γ-globin expression. When treated with hydroxyurea or inhibitors to histone deacetylases, DNA methyltransferases, or the histone demethylase lysine specific demethylase 1 (LSD1), β-Yac ESREs significantly increase their γ-globin expression, demonstrating their utility for studying agents that influence maturational globin switching. β-yac ESREs were further used to characterize the secondary effects of LSD1 inhibition on erythroid maturation, with inhibition of LSD1 resulting in altered cell and nuclear size, prolonged Kit expression, and decreased rates of enucleation consistent with impaired maturation. Taken together, these studies demonstrate that β-yac ESREs have significant utility for identifying modulators of maturational globin switching as well as for studying the broader role of those modulators in erythroid maturation.
珠蛋白基因的调控发生在成熟红细胞的背景下,在此过程中染色质结构和基因表达发生重大变化。很少有模型系统可用于研究红细胞成熟背景下的珠蛋白基因调控。高度自我更新的红细胞(ESREs)是一种源自鼠胎儿肝脏或卵黄囊的未转化的红细胞成熟模型。成像流式细胞术和 RNA-seq 研究表明,ESREs在功能和分子水平上模拟了红细胞成熟。为了解决需要一种也能重现人类珠蛋白开关的模型系统的问题,ESREs是从转基因小鼠中衍生而来的,这些小鼠携带完整的人类β-珠蛋白基因座(β-yac ESREs)。β-yac ESREs 从转基因的人类基因座表达β-珠蛋白,γ-珠蛋白表达极少。当用羟基脲或组蛋白去乙酰化酶、DNA 甲基转移酶或组蛋白去甲基酶赖氨酸特异性去甲基酶 1(LSD1)抑制剂处理时,β-Yac ESREs 显著增加其 γ-珠蛋白表达,证明它们可用于研究影响成熟珠蛋白开关的药物。进一步使用β-yac ESREs 来表征 LSD1 抑制对红细胞成熟的次要影响,抑制 LSD1 导致细胞和核大小改变、Kit 表达延长以及去核率降低,与成熟受损一致。综上所述,这些研究表明,β-yac ESREs 在鉴定成熟珠蛋白开关的调节剂以及研究这些调节剂在红细胞成熟中的更广泛作用方面具有重要的应用价值。