Efficient in vivo targeting of epidermal stem cells by early gestational intraamniotic injection of lentiviral vector driven by the keratin 5 promoter
Efficient in vivo targeting of epidermal stem cells by early gestational intraamniotic injection of lentiviral vector driven by the keratin 5 promoter
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DOI：
10.1038/sj.mt.6300332
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发表时间：
2008-01-01
影响因子：
12.4
通讯作者：
Flake, Alan W.
中科院分区：
文献类型：
--
作者：
Endo, Masayuki;Zoltick, Philip W.;Flake, Alan W.
目前，还没有有效的体内基因转移到皮肤干细胞的方法。在这项研究中，我们假设在怀孕早期，特定的表皮干细胞群体可能可以进行基因转移。为了验证这一假设，我们将编码绿色荧光蛋白标志基因的慢病毒载体在胚胎第8天到第12天的早期发育阶段将由巨细胞病毒启动子或角蛋白5(K5)启动子驱动的慢病毒载体注射到小鼠羊膜腔中。这导致绿色荧光蛋白(GFP)在皮肤毛囊中的基底表皮干细胞和隆起细胞中持续表达。干细胞群的转导依赖于发育阶段，并从所有皮肤成分中GFP表达的持续时间延长到成年期来证实。此外，转导的干细胞群体对创伤后的再生信号有反应，并积极参与伤口愈合。最后，我们量化了转导的表皮干细胞的比例，以及与所使用的启动子相关的转导的分布，证实了K5启动子的效率提高。这种简单的方法可能在我们的皮肤基因功能研究中有生物学应用，也许未来还会应用于皮肤病的临床治疗。
At the present time, no efficient in vivo method for gene transfer to skin stem cells exists. In this study, we hypothesized that early in gestation, specific epidermal stem cell populations may be accessible for gene transfer. To test this hypothesis, we injected lentiviral vectors encoding the green fluorescence protein marker gene driven by either the cytomegalovirus promoter or the keratin 5 (K5) promoter into the murine amniotic space at early developmental stages between embryonic days 8 and 12. This resulted in sustained green fluorescent protein (GFP) expression in both basal epidermal stem cells and bulge cells in the hair follicles of the skin. Transduction of stem cell populations was dependent on the developmental stage, and confirmed by the prolonged duration of GFP expression in all skin elements into adulthood. In addition, transduced stem cell populations responded to regenerative signals after wounding and actively participated in wound healing. Finally, we quantified the fraction of epidermal stem cells transduced, and the distribution of transduction related to the promoters utilized, confirming improved efficiency with the K5 promoter. This simple approach has possible biological applications in our study of gene functions in skin, and perhaps future clinical applications for treatment of skin based disorders.