Extensively self-renewing erythroblasts derived from transgenic β-yac mice is a novel model system for studying globin switching and erythroid maturation.
Extensively self-renewing erythroblasts derived from transgenic β-yac mice is a novel model system for studying globin switching and erythroid maturation.
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DOI：
10.1016/j.exphem.2014.03.006
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发表时间：
2014-07
影响因子：
2.6
通讯作者：
Steiner LA
中科院分区：
文献类型：
--
作者：
Getman M;England SJ;Malik J;Peterson K;Palis J;Steiner LA
珠蛋白基因调控发生在成熟的红系细胞的背景下，其正在经历染色质结构和基因表达的显著变化。有几个可用的模型系统，促进珠蛋白基因调控的红系成熟的背景下的研究。广泛自我更新成红细胞（ESRE）是来源于小鼠胎肝或卵黄囊的红系成熟的非转化模型。成像流式细胞术和RNA-seq研究证明ESRE在功能上和分子上模拟红系成熟。为了解决对也重现人球蛋白转换的模型系统的需要，ESRE来源于完整人β-球蛋白基因座的转基因小鼠（β-yac ESRE）。β-yac ESRE表达来自转基因人基因座的β-珠蛋白，具有最小的γ-珠蛋白表达。当用羟基脲或组蛋白脱乙酰酶、DNA甲基转移酶或组蛋白脱甲基酶赖氨酸特异性脱甲基酶1（LSD 1）的抑制剂处理时，β-Yac ESRE显著增加其γ-珠蛋白表达，证明其用于研究影响成熟珠蛋白转换的试剂的效用。β-yac ESRE进一步用于表征LSD 1抑制对红系成熟的次级效应，LSD 1的抑制导致细胞和核大小改变、Kit表达延长以及与成熟受损一致的去核速率降低。总之，这些研究表明β-yac ESRE对于鉴定成熟球蛋白转换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具有重要的实用性。
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.
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