Human artificial chromosome (HAC) vector provides long-term therapeutic transgene expression in normal human primary fibroblasts
Human artificial chromosome (HAC) vector provides long-term therapeutic transgene expression in normal human primary fibroblasts
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DOI：
10.1038/sj.gt.3302483
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发表时间：
2005-05-01
期刊：
影响因子：
5.1
通讯作者：
Tomizuka, K
中科院分区：
文献类型：
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作者：
Kakeda, M;Hiratsuka, M;Tomizuka, K
人类人工染色体(HAC)从宿主染色体中自由分离，在治疗性基因传递中确保基因表达的安全性和持续时间是潜在的有用的。然而，完整的HAC向细胞的低转移效率阻碍了使用正常人类原代细胞的研究，而原代细胞是体外基因治疗的主要靶点。为了阐明HAC作为基因治疗载体的潜力，我们研究了将HAC载体通过微细胞介导的染色体转移(MMCT)导入正常原代人成纤维细胞(HPFS)中。我们证明了含有结构定义的额外HAC载体的细胞遗传学正常的HPFS的产生。HAc载体在HPFS中稳定存在，HAc在宿主染色体上没有易位。我们还在其中实现了至少12周的人促红细胞生成素的长期生产。这些结果揭示了HAC作为新的选择来绕过传统基因治疗载体的问题的能力。
Human artificial chromosomes (HACs) segregating freely from host chromosomes are potentially useful to ensure both safety and duration of gene expression in therapeutic gene delivery. However, low transfer efficiency of intact HACs to the cells has hampered the studies using normal human primary cells, the major targets for ex vivo gene therapy. To elucidate the potential of HACs to be vectors for gene therapy, we studied the introduction of the HAC vector, which is reduced in size and devoid of most expressed genes, into normal primary human fibroblasts (hPFs) with microcell-mediated chromosome transfer (MMCT). We demonstrated the generation of cytogenetically normal hPFs harboring the structurally defined and extra HAC vector. This introduced HAC vector was retained stably in hPFs without translocation of the HAC on host chromosomes. We also achieved the long-term production of human erythropoietin for at least 12 weeks in them. These results revealed the ability of HACs as novel options to circumvent issues of conventional vectors for gene therapy.