Targeting expression of a transgene to the airway surface epithelium using a ciliated cell-specific promoter
Targeting expression of a transgene to the airway surface epithelium using a ciliated cell-specific promoter
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DOI：
10.1016/s1525-0016(03)00221-1
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发表时间：
2003-10-01
影响因子：
12.4
通讯作者：
O'Neal, WK
中科院分区：
文献类型：
--
作者：
Ostrowski, LE;Hutchins, JR;O'Neal, WK
许多正在研究的用于基因治疗的载体利用病毒启动子或来自普遍表达的基因的启动子(例如，CMV、β-肌动蛋白)。这些启动子在许多类型的细胞中都是活跃的，通常会导致高水平的转基因表达。然而，使用这些启动子进行囊性纤维化(CF)的基因治疗可能会导致在正常情况下不合成该蛋白的细胞中高水平表达CFTR，从而产生不良影响。相反，含有纤毛细胞特异性启动子并输送到肺部的载体只会在排列在呼吸道表面的纤毛细胞中发挥作用。纤毛细胞表达cftr，并与正常情况下受cftr调节的呼吸道表面液体直接接触。为了开发纤毛细胞特异性启动子用于CF基因治疗，我们鉴定了FOXJ1基因的启动子区域，FOXJ1基因是纤毛细胞分化所需的转录因子。将人FOXJ1启动子片段插入绿色荧光蛋白表达盒，用于制备转基因小鼠。转基因阳性动物在气管、支气管和鼻腔上皮的纤毛细胞中有较强的EGFP表达。我们的结果表明，FOXJ1启动子区域内的元件足以将转基因靶向表达到纤毛细胞，并可能用于CF的基因治疗。
Many of the vectors being investigated for gene therapy utilize viral promoters or promoters from ubiquitously expressed genes (e.g., CMV, beta-actin). These promoters are active in many cell types and generally result in high levels of transgene expression. However, the use of these promoters for gene therapy of cystic fibrosis (CF) may produce undesirable effects by directing high levels of CFTR expression in cells that normally do not synthesize this protein. In contrast, a vector containing a ciliated cell-specific promoter and delivered to the lung would be active only in the ciliated cells that line the surface of the airways. Ciliated cells express CFTR and are in direct contact with the airway surface liquid normally regulated by CFTR. To develop a ciliated cell-specific promoter for CF gene therapy, we have characterized the promoter region of the FOXJ1 gene, a transcription factor required for ciliated cell differentiation. A fragment of the human FOXJ1 promoter region was inserted into an EGFP expression cassette and used to produce transgenic mice. Transgene-positive animals demonstrated strong EGFP expression in the ciliated cells of tracheal, bronchial, and nasal epithelium. Our results demonstrate that elements within the FOXJ1 promoter region are sufficient to target expression of transgenes to ciliated cells and may be useful for gene therapy of CF.