Production of beta-globin and adult hemoglobin following G418 treatment of erythroid precursor cells from homozygous beta(0)39 thalassemia patients.
Production of beta-globin and adult hemoglobin following G418 treatment of erythroid precursor cells from homozygous beta(0)39 thalassemia patients.
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对纯合 β(0)39 地中海贫血患者的红系前体细胞进行 G418 处理后，产生 β-珠蛋白和成人血红蛋白。
DOI：
10.1002/ajh.21539
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发表时间：
2009
影响因子：
12.8
通讯作者：
Gambari
中科院分区：
文献类型：
--
作者：
Salvatori,Francesca;Breveglieri,Giulia;Zuccato,Cristina;Finotti,Alessia;Bianchi,Nicoletta;Borgatti,Monica;Feriotto,Giordana;Destro,Federica;Canella,Alessandro;Brognara,Eleonora;Lampronti,Ilaria;Breda,Laura;Rivella,Stefano;Gambari
在几种类型的地中海贫血(包括β039-地中海贫血)中，终止密码子突变导致提前翻译终止，并通过无义介导的衰变导致mRNA失稳。药物(例如氨基糖苷类)可以被设计用来抑制早产，导致核糖体通读。这些发现为开发治疗这种疾病的药理学方法带来了新的希望。然而，氨基糖苷类药物对携带β-地中海贫血终止突变的珠蛋白基因的影响尚未被研究。在这项研究中，我们使用了含有β039-地中海贫血珠蛋白基因的慢病毒载体，该基因受β-珠蛋白启动子和LCR盒的控制。我们通过荧光激活细胞分选分析证明，表达β039-地中海贫血珠蛋白基因的K562细胞克隆经G418处理后产生β-珠蛋白。更重要的是，经过流式细胞仪和高效液相分析，来自β039-地中海贫血患者的红系前体细胞经G418处理后能够产生β-珠蛋白和成人血红蛋白。这项研究强烈表明，核糖体通读应该被认为是开发治疗由终止密码子突变引起的β0地中海贫血的实验策略的一种策略。上午好。J.Hematol.，2009年。©2009威利-利斯，Inc.
In several types of thalassemia (including β039‐thalassemia), stop codon mutations lead to premature translation termination and to mRNA destabilization through nonsense‐mediated decay. Drugs (for instance aminoglycosides) can be designed to suppress premature termination, inducing a ribosomal readthrough. These findings have introduced new hopes for the development of a pharmacologic approach to the cure of this disease. However, the effects of aminoglycosides on globin mRNA carrying β‐thalassemia stop mutations have not yet been investigated. In this study, we have used a lentiviral construct containing the β039‐thalassemia globin gene under control of the β‐globin promoter and a LCR cassette. We demonstrated by fluorescence‐activated cell sorting (FACS) analysis the production of β‐globin by K562 cell clones expressing the β039‐thalassemia globin gene and treated with G418. More importantly, after FACS and high‐performance liquid chromatography (HPLC) analyses, erythroid precursor cells from β039‐thalassemia patients were demonstrated to be able to produce β‐globin and adult hemoglobin after treatment with G418. This study strongly suggests that ribosomal readthrough should be considered a strategy for developing experimental strategies for the treatment of β0‐thalassemia caused by stop codon mutations. Am. J. Hematol., 2009. © 2009 Wiley‐Liss, Inc.