Retroviral transfer of a human beta-globin/delta-globin hybrid gene linked to beta locus control region hypersensitive site 2 aimed at the gene therapy of sickle cell disease.
Retroviral transfer of a human beta-globin/delta-globin hybrid gene linked to beta locus control region hypersensitive site 2 aimed at the gene therapy of sickle cell disease.
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与 β 基因座控制区超敏位点 2 连接的人 β-珠蛋白/δ-珠蛋白杂合基因的逆转录病毒转移旨在镰状细胞病的基因治疗。
DOI：
10.1073/pnas.92.7.3014
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发表时间：
1995
影响因子：
11.1
通讯作者：
Leboulch,P
中科院分区：
文献类型：
--
作者：
Takekoshi,KJ;Oh,YH;Westerman,KW;London,IM;Leboulch,P
人的丙种球蛋白链和β-珠蛋白链此前已被确定为对S的血红蛋白聚合有强烈的抑制作用，而β-珠蛋白链只起到中等的抗病作用。然而，与β-珠蛋白相反，在成人红系细胞中，γ-珠蛋白和β-珠蛋白的表达水平通常很低。我们报道了β-珠蛋白/β-珠蛋白杂合基因--β/β-镰状细胞抑制因子1(β/β-SCI1)的设计及其逆转录病毒介导的基因转移。β/Delta-SCI1编码基因保留了人类β-珠蛋白基因的整体结构，同时包含了先前发现的与其增强的抗病特性有关的三角洲链的特定氨基酸残基。为了在成人红细胞中实现β/β-SCI1的高水平表达，该杂交基因被置于人β-珠蛋白启动子和人β基因位点控制区的DNase I超敏部位2的转录调控下。产生了高滴度的逆转录病毒，并在感染细胞中实现了稳定的前病毒传播。在感染的二甲基亚砜诱导的小鼠红白血病细胞中，β/β-SCI1基因的基因表达水平接近内源性小鼠β-珠蛋白基因表达水平的85%，这证明在成年红系细胞中获得了高水平的基因表达。在镰状细胞病的转基因动物模型中进一步评估这一策略，应该评估其对人类患者的基因治疗的有效性。
Human gamma-globin and delta-globin chains have been previously identified as strong inhibitors of the polymerization of hemoglobin S, in contrast to the beta-globin chain, which exerts only a moderate antisickling effect. However, gamma-globin and delta-globin are normally expressed at very low levels in adult erythroid cells, in contrast to beta-globin. We report the design of a beta-globin/delta-globin hybrid gene, beta/delta-sickle cell inhibitor 1 (beta/delta-SCI1) and its transduction by retrovirus-mediated gene transfer. The beta/delta-SCI1-encoding gene retains the overall structure of the human beta-globin gene, while incorporating specific amino acid residues from the delta chain previously found responsible for its enhanced antisickling properties. To achieve high expression levels of beta/delta-SCI1 in adult erythrocytes, the hybrid gene was placed under the transcriptional control of the human beta-globin promoter and the DNase I hypersensitive site 2 of the human beta locus control region. High-titer retroviruses were generated, and stable proviral transmission was achieved in infected cells. The mRNA expression levels of the beta/delta-SCI1 gene in infected, dimethyl sulfoxide-induced murine erythroleukemia cells approached 85% of the endogenous murine beta maj-globin mRNA, on a per gene basis, evidence that high gene expression levels were achieved in adult erythroid cells. Further evaluation of this strategy in transgenic animal models of sickle cell disease should assess its efficacy for the gene therapy of human patients.