May C, Rivella S, Callegari J, Heller G, Gaensler K M, Luzzatto L, Sadelain M
Department of Human Genetics, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
Nature. 2000 Jul 6;406(6791):82-6. doi: 10.1038/35017565.
The stable introduction of a functional beta-globin gene in haematopoietic stem cells could be a powerful approach to treat beta-thalassaemia and sickle-cell disease. Genetic approaches aiming to increase normal beta-globin expression in the progeny of autologous haematopoietic stem cells might circumvent the limitations and risks of allogeneic cell transplants. However, low-level expression, position effects and transcriptional silencing hampered the effectiveness of viral transduction of the human beta-globin gene when it was linked to minimal regulatory sequences. Here we show that the use of recombinant lentiviruses enables efficient transfer and faithful integration of the human beta-globin gene together with large segments of its locus control region. In long-term recipients of unselected transduced bone marrow cells, tetramers of two murine alpha-globin and two human betaA-globin molecules account for up to 13% of total haemoglobin in mature red cells of normal mice. In beta-thalassaemic heterozygous mice higher percentages are obtained (17% to 24%), which are sufficient to ameliorate anaemia and red cell morphology. Such levels should be of therapeutic benefit in patients with severe defects in haemoglobin production.
在造血干细胞中稳定引入功能性β-珠蛋白基因可能是治疗β地中海贫血和镰状细胞病的有效方法。旨在增加自体造血干细胞后代中正常β-珠蛋白表达的基因方法可能会规避异基因细胞移植的局限性和风险。然而,当人类β-珠蛋白基因与最小调控序列相连时,低水平表达、位置效应和转录沉默阻碍了病毒转导该基因的有效性。在此我们表明,使用重组慢病毒能够实现人类β-珠蛋白基因与其大片段基因座控制区的高效转移和忠实整合。在未选择的转导骨髓细胞的长期受体中,两个小鼠α-珠蛋白和两个人类βA-珠蛋白分子的四聚体在正常小鼠成熟红细胞的总血红蛋白中占比高达13%。在β地中海贫血杂合小鼠中,该比例更高(17%至24%),足以改善贫血和红细胞形态。这样的水平对血红蛋白产生严重缺陷的患者应具有治疗益处。