Topical colchicine selection of keratinocytes transduced with the multidrug resistance gene (MDR1) can sustain and enhance transgene expression in vivo.
Topical colchicine selection of keratinocytes transduced with the multidrug resistance gene (MDR1) can sustain and enhance transgene expression in vivo.
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对转导多药耐药基因 (MDR1) 的角质形成细胞进行局部秋水仙碱选择可以维持和增强体内转基因表达。
DOI：
10.1073/pnas.192247899
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发表时间：
2002
影响因子：
11.1
通讯作者：
Vogel,JC
中科院分区：
文献类型：
--
作者：
Pfutzner,W;Terunuma,A;Tock,CL;Snead,EK;Kolodka,TM;Gottesman,MM;Taichman,L;Vogel,JC
对于皮肤基因治疗来说，在相当比例的角质形成细胞 (KC) 中实现长时间的高水平基因表达是很困难的，因为我们无法选择性地靶向 KC 干细胞。我们现在证明，局部秋水仙碱治疗可用于体内选择转导多药耐药基因 (MDR1) 的 KC 祖细胞。当将含有 MDR1 转导的 KC 的人类皮肤等同物移植到免疫功能低下的小鼠身上时，与载体处理的对照组相比，局部秋水仙碱治疗显着增加了表达 MDR1 的 KC 的百分比（7 倍），持续时间长达 24 周。局部秋水仙碱治疗还显着增加了个体 KC 中 MDR1 蛋白的表达量。此外，MDR1转基因拷贝数的定量实时PCR分析表明，局部秋水仙碱治疗选择并富集了皮肤中含有并表达MDR1的KC祖细胞。对于临床皮肤基因治疗应用，这种体内选择方法有望通过将其表达与 MDR1 选择标记基因联系起来，增强 KC 中所需治疗基因的持续时间和表达水平。
For skin gene therapy, achieving prolonged high-level gene expression in a significant percentage of keratinocytes (KC) is difficult because we cannot selectively target KC stem cells. We now demonstrate that topical colchicine treatment can be used to select,in vivo, KC progenitor cells transduced with the multidrug resistance gene (MDR1). When human skin equivalents containingMDR1-transduced KC were grafted onto immunocompromised mice, topical colchicine treatments significantly increased (7-fold) the percentage of KC expressing MDR1, compared to vehicle-treated controls, for up to 24 wk. Topical colchicine treatment also significantly enhanced the amount of MDR1 protein expressed in individual KC. Furthermore, quantitative real-time PCR analysis ofMDR1transgene copy number demonstrates that topical colchicine treatment selects and enriches for KC progenitor cells in the skin that contain and express MDR1. For clinical skin gene therapy applications, thisin vivoselection approach promises to enhance both the duration and expression level of a desired therapeutic gene in KC, by linking its expression to theMDR1selectable marker gene.