Human miR223 Promoter as a Novel Myelo-Specific Promoter for Chronic Granulomatous Disease Gene Therapy
Human miR223 Promoter as a Novel Myelo-Specific Promoter for Chronic Granulomatous Disease Gene Therapy
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DOI：
10.1089/hgtb.2012.157
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发表时间：
2013-06-01
影响因子：
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通讯作者：
Siler, Ulrich
中科院分区：
文献类型：
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作者：
Brendel, Christian;Haenseler, Walther;Siler, Ulrich
将转基因表达靶向到特定的造血细胞系可以提高逆转录病毒载体在基因治疗应用中的安全性。慢性肉芽肿病（CGD）是一种吞噬细胞的缺陷，可以通过基因治疗来控制，使用靶向表达骨髓细胞的逆转录病毒载体。在这种情况下，我们分析了人类miR223启动子的骨髓特异性，该启动子在骨髓分化过程中被强烈上调，以驱动CGD小鼠模型和原代患者来源细胞中p47（phox）和gp91（phox）的骨髓限制性表达。miR223启动子将自我灭活(SIN) γ和慢病毒载体中的p47（phox）、gp91（phox）和绿色荧光蛋白（GFP）的表达限制在粒细胞和巨噬细胞中，仅在淋巴细胞或造血干细胞和祖细胞中边缘性表达。此外，将基因转移到p47（phox）患者的原代CD34+细胞中，然后体外分化为中性粒细胞，通过miR223启动子介导的p47（phox）表达恢复大肠杆菌的杀伤活性。这些结果表明，miR223启动子作为SIN基因治疗载体中的内部启动子能够有效地纠正造血祖细胞中的CGD表型，而活性可以忽略不计，从而限制插入性肿瘤发生和克隆优势发展的风险。
Targeting transgene expression to specific hematopoietic cell lineages could contribute to the safety of retroviral vectors in gene therapeutic applications. Chronic granulomatous disease (CGD), a defect of phagocytic cells, can be managed by gene therapy, using retroviral vectors with targeted expression to myeloid cells. In this context, we analyzed the myelospecificity of the human miR223 promoter, which is known to be strongly upregulated during myeloid differentiation, to drive myeloid-restricted expression of p47(phox) and gp91(phox) in mouse models of CGD and in primary patient-derived cells. The miR223 promoter restricted the expression of p47(phox), gp91(phox), and green fluorescent protein (GFP) within self-inactivating (SIN) gamma-and lentiviral vectors to granulocytes and macrophages, with only marginal expression in lymphocytes or hematopoietic stem and progenitor cells. Furthermore, gene transfer into primary CD34+ cells derived from a p47(phox) patient followed by ex vivo differentiation to neutrophils resulted in restoration of Escherichia coli killing activity by miR223 promoter-mediated p47(phox) expression. These results indicate that the miR223 promoter as an internal promoter within SIN gene therapy vectors is able to efficiently correct the CGD phenotype with negligible activity in hematopoietic progenitors, thereby limiting the risk of insertional oncogenesis and development of clonal dominance.