DNA damage and gene therapy of xeroderma pigmentosum, a human DNA repair-deficient disease
DNA damage and gene therapy of xeroderma pigmentosum, a human DNA repair-deficient disease
复制标题
DOI：
10.1016/j.mrfmmm.2014.08.007
复制
发表时间：
2015-06-01
影响因子：
2.3
通讯作者：
Sarasin, Alain
中科院分区：
文献类型：
--
作者：
Dupuy, Aurelie;Sarasin, Alain
着色性干皮病（XP）是一种遗传性疾病，其特征是对紫外线过敏，并且在暴露的身体部位诱发皮肤癌的风险非常高。这种综合征是由核苷酸切除修复基因的突变引起的。除了完全保护免受所有类型的紫外线辐射之外，这些患者无法治愈。我们综述了各种技术来补充或纠正XP细胞中的遗传缺陷。我们特别开发了在XPC野生型序列存在下修复双链断裂（DSB）期间使用同源重组途径的保真度的XP-C皮肤细胞的校正。我们使用工程化的核酸酶（大范围核酸酶或TALE核酸酶）来诱导位于待校正突变的90 bp处的DSB。在含有一长段同源野生型XPC序列的修复基质存在下表达特异性TALE核酸酶允许我们成功地基因校正在许多北非XP患者中发现的原始TG缺失。一些工程化核酸酶对表观遗传修饰敏感，如胞嘧啶甲基化。在要校正的甲基化序列的情况下，应设想修饰的核酸酶或全基因组的去甲基化。总的来说，我们表明，专门设计的TALE-核酸酶允许我们纠正XPC基因中的2bp缺失，导致患者的细胞精通DNA修复并显示正常的UV敏感性。修正后的基因仍然在人类基因组中的相同位置，并处于其生理启动子的调控下。这一结果是XP患者基因治疗的第一步。(C)2014爱思唯尔有限公司版权所有。
Xeroderma pigmentosum (XP) is a genetic disease characterized by hypersensitivity to ultra-violet and a very high risk of skin cancer induction on exposed body sites. This syndrome is caused by germinal mutations on nucleotide excision repair genes. No cure is available for these patients except a complete protection from all types of UV radiations. We reviewed the various techniques to complement or to correct the genetic defect in XP cells. We, particularly, developed the correction of XP-C skin cells using the fidelity of the homologous recombination pathway during repair of double-strand break (DSB) in the presence of XPC wild type sequences. We used engineered nucleases (meganuclease or TALE nuclease) to induce a DSB located at 90 bp of the mutation to be corrected. Expression of specific TALE nuclease in the presence of a repair matrix containing a long stretch of homologous wild type XPC sequences allowed us a successful gene correction of the original TG deletion found in numerous North African XP patients. Some engineered nucleases are sensitive to epigenetic modifications, such as cytosine methylation. In case of methylated sequences to be corrected, modified nucleases or demethylation of the whole genome should be envisaged. Overall, we showed that specifically-designed TALE-nuclease allowed us to correct a 2 bp deletion in the XPC gene leading to patient's cells proficient for DNA repair and showing normal UV-sensitivity. The corrected gene is still in the same position in the human genome and under the regulation of its physiological promoter. This result is a first step toward gene therapy in XP patients. (C) 2014 Elsevier B.V. All rights reserved.