Wu X, Burgess S M
Laboratory of Molecular Technology, Scientific Application International Inc., National Cancer Institute at Frederick, 915 Tollhouse Ave., Frederick, Maryland 21701, USA.
Cell Mol Life Sci. 2004 Oct;61(19-20):2588-96. doi: 10.1007/s00018-004-4206-9.
When a retrovirus infects a cell, its RNA genome is reverse transcribed into a double-stranded DNA, which is then permanently integrated into the host chromosome. Integration is one of the essential steps in the retroviral life cycle. Many transposable elements also move around and integrate into the host genome as part of their life cycle, some through RNA intermediates and some through 'cut and paste' mechanisms. Integration of retroviruses and transposable elements into 'sensitive areas' of the genome can cause irreparable damage. On the other hand, because of their ability to integrate permanently, and the relatively efficient rates of transgenesis, retroviruses and transposable elements are widely used as gene delivery tools in basic research and gene therapy trials. Recent events in gene therapy treatments for X-linked severe combined immunity deficiencies (X-SCID) have highlighted both the promise and some of the risks involved with utilizing retroviruses. Nine of 11 children were successfully treated for X-SCID using a retrovirus carrying the gene mutated in this disease. However, later two of these children developed leukemias because of retroviral integrations in the putative oncogene LMO2 [1]. A third child has also been demonstrated to have an integration in LMO2, but is as of yet nonsymptomatic [2]. It is a bit difficult to explain the high frequency of integrations into the same gene using a random model of retroviral integration, and there has been evidence for decades that retroviral integrations may not be random. But the data were somewhat limited in their power to determine the precise nature of the integration biases. The completion of the human genome sequence coupled with sensitive polymerase chain reaction techniques and an ever-decreasing cost of sequencing has given a powerful new tool to the study of integration site selection. In this review, we describe the findings from several recent global surveys of target site selection by retroviruses and transposable elements, and discuss the possible ramifications of these findings to both mechanisms of action and to the use of these elements as gene therapy vectors.
当逆转录病毒感染细胞时,其RNA基因组会被逆转录成双链DNA,然后永久性地整合到宿主染色体中。整合是逆转录病毒生命周期中的关键步骤之一。许多转座元件在其生命周期中也会移动并整合到宿主基因组中,一些通过RNA中间体,一些通过“剪切粘贴”机制。逆转录病毒和转座元件整合到基因组的“敏感区域”会造成无法修复的损害。另一方面,由于它们具有永久整合的能力以及相对高效的转基因效率,逆转录病毒和转座元件在基础研究和基因治疗试验中被广泛用作基因传递工具。最近在X连锁严重联合免疫缺陷症(X-SCID)基因治疗中的事件凸显了利用逆转录病毒的前景和一些风险。11名儿童中有9名使用携带该疾病突变基因的逆转录病毒成功治疗了X-SCID。然而,后来其中两名儿童因逆转录病毒整合到假定的致癌基因LMO2中而患上白血病[1]。第三名儿童也被证明LMO2中有整合,但目前尚无症状[2]。使用逆转录病毒整合的随机模型来解释整合到同一基因中的高频率现象有点困难,而且几十年来一直有证据表明逆转录病毒整合可能不是随机的。但这些数据在确定整合偏差的确切性质方面的能力有些有限。人类基因组序列的完成,加上灵敏的聚合酶链反应技术以及测序成本的不断降低,为整合位点选择的研究提供了一个强大的新工具。在这篇综述中,我们描述了最近几项关于逆转录病毒和转座元件靶位点选择的全球调查结果,并讨论了这些发现对作用机制以及将这些元件用作基因治疗载体的可能影响。