A zinc-finger transcriptional activator designed to interact with the gamma-globin gene promoters enhances fetal hemoglobin production in primary human adult erythroblasts.
A zinc-finger transcriptional activator designed to interact with the gamma-globin gene promoters enhances fetal hemoglobin production in primary human adult erythroblasts.
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DOI：
10.1182/blood-2009-08-240556
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发表时间：
2010-04
期刊：
影响因子：
20.3
通讯作者：
A. Wilber;Ulrich Tschulena;Phillip W. Hargrove;Yoon-Sang Kim;D. Persons;C. Barbas;A. Nienhuis
中科院分区：
文献类型：
--
作者：
A. Wilber;Ulrich Tschulena;Phillip W. Hargrove;Yoon-Sang Kim;D. Persons;C. Barbas;A. Nienhuis
胎儿血红蛋白（HbF）是β-地中海贫血和镰状细胞贫血严重程度的有效遗传修饰剂。我们使用了一个体外培养的人红细胞生成模型，其中晚期成红细胞直接来源于人CD 34（+）造血细胞，以评估HbF的产生。该系统根据起源细胞来源的发育阶段再现珠蛋白基因的表达。当培养成人外周血CD 34（+）细胞时，HbF的背景水平为2%或更低。培养的细胞在暴露于载体颗粒48 - 72小时之间时，很容易被慢病毒载体转导。在可能增强胎儿血红蛋白产生的遗传元件中，有一种人工锌指转录因子GG 1-VP 64，旨在与近端γ-珠蛋白基因启动子相互作用。我们的数据显示，在相对弱的红细胞特异性启动子的控制下，慢病毒介导的GG 1-VP 64的强制表达在源自成人CD 34（+）细胞的成红细胞中诱导显著量的HbF（高达20%），而不改变其红细胞成熟的能力，并且仅适度减少转导后在培养物中积累的细胞总数。这些观察结果表明，在β-地中海贫血或镰状细胞性贫血患者中，HbF的序列特异性增强的潜力。
Fetal hemoglobin (HbF) is a potent genetic modifier of the severity of beta-thalassemia and sickle cell anemia. We used an in vitro culture model of human erythropoiesis in which late-stage erythroblasts are derived directly from human CD34(+) hematopoietic cells to evaluate HbF production. This system recapitulates expression of globin genes according to the developmental stage of the originating cell source. When cytokine-mobilized peripheral blood CD34(+) cells from adults were cultured, background levels of HbF were 2% or less. Cultured cells were readily transduced with lentiviral vectors when exposed to vector particles between 48 and 72 hours. Among the genetic elements that may enhance fetal hemoglobin production is an artificial zinc-finger transcription factor, GG1-VP64, designed to interact with the proximal gamma-globin gene promoters. Our data show that lentiviral-mediated, enforced expression of GG1-VP64 under the control of relatively weak erythroid-specific promoters induced significant amounts of HbF (up to 20%) in erythroblasts derived from adult CD34(+) cells without altering their capacity for erythroid maturation and only modestly reducing the total numbers of cells that accumulate in culture after transduction. These observations demonstrate the potential for sequence-specific enhancement of HbF in patients with beta-thalassemia or sickle cell anemia.