Lineage- and stage-restricted lentiviral vectors for the gene therapy of chronic granulomatous disease
Lineage- and stage-restricted lentiviral vectors for the gene therapy of chronic granulomatous disease
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DOI：
10.1038/gt.2011.65
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发表时间：
2011-11-01
期刊：
影响因子：
5.1
通讯作者：
Trono, D.
中科院分区：
文献类型：
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作者：
Barde, I.;Laurenti, E.;Trono, D.
插入突变是整合载体基因治疗的一个严重副作用。然而，虽然在干细胞中生长促进基因的不受控制的激活可以预见会导致肿瘤过程，但如果载体转录活性可以限制在完全分化的细胞中，这是不太可能的。仅需要在成熟细胞中进行表型校正的疾病提供了这样的机会，前提是可以适当地定制谱系/阶段限制系统。在这项研究中，我们遵循这一推理，设计慢病毒载体的基因治疗慢性肉芽肿病（CGD），免疫缺陷，由于损失的烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶在吞噬细胞，最常见的继发于突变gp 91（phox）。使用自失活HIV 1衍生载体作为背景，我们首先从最小gp 91（phox）启动子表达增强型绿色荧光蛋白（eGFP），添加各种天然或合成转录调控元件以促进特异性和效力。通过移植或通过慢病毒转基因评估所得载体，寻找赋予成熟吞噬细胞强和特异性表达的组合。最有希望的载体被修饰以表达gp 91（phox）并用于治疗CGD小鼠。在给药动物的粒细胞中记录了NADPH活性的高水平恢复。我们认为这种谱系特异性慢病毒载体是CGD基因治疗的合适候选载体。Gene Therapy（2011）18，1087-1097; doi：10.1038/gt.2011.65; 2011年5月5日在线发表
Insertional mutagenesis represents a serious adverse effect of gene therapy with integrating vectors. However, although uncontrolled activation of growth-promoting genes in stem cells can predictably lead to oncological processes, this is far less likely if vector transcriptional activity can be restricted to fully differentiated cells. Diseases requiring phenotypic correction only in mature cells offer such an opportunity, provided that lineage/stage-restricted systems can be properly tailored. In this study, we followed this reasoning to design lentiviral vectors for the gene therapy of chronic granulomatous disease (CGD), an immune deficiency due a loss of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in phagocytes, most often secondary to mutations in gp91(phox). Using self-inactivating HIV1-derived vectors as background, we first expressed enhanced green fluorescent protein (eGFP) from a minimal gp91(phox) promoter, adding various natural or synthetic transcriptional regulatory elements to foster both specificity and potency. The resulting vectors were assessed either by transplantation or by lentiviral transgenesis, searching for combinations conferring strong and specific expression into mature phagocytic cells. The most promising vector was modified to express gp91(phox) and used to treat CGD mice. High-level restoration of NADPH activity was documented in granulocytes from the treated animals. We propose that this lineage-specific lentiviral vector is a suitable candidate for the gene therapy of CGD. Gene Therapy (2011) 18, 1087-1097; doi:10.1038/gt.2011.65; published online 5 May 2011