Targeting transgene expression to airway epithelia and submucosal glands, prominent sites of human CFTR expression
Targeting transgene expression to airway epithelia and submucosal glands, prominent sites of human CFTR expression
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DOI：
10.1006/mthe.2000.0135
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发表时间：
2000-10-01
影响因子：
12.4
通讯作者：
Hu, J
中科院分区：
文献类型：
--
作者：
Chow, YH;Plumb, J;Hu, J
将治疗基因表达靶向受疾病影响的组织是有效且安全的基因治疗的重要组成部分。出生后，人肺中的 CFTR 基因表达主要集中在上呼吸道内壁的上皮细胞中，尤其是粘膜下腺的导管和浆液小管中。我们基于人类细胞角蛋白 18 基因的 DNA 控制元件开发了 K18 表达盒。对转基因小鼠的时间和空间分析表明，该表达盒将转基因表达靶向几乎所有表达 CFTR 的细胞类型。气道上皮早在胎龄 11.5 天时就开始表达，并持续到转基因小鼠的成年期。在这些成年小鼠中，报告基因的表达模式与人类细胞角蛋白 18 和人类 CFTR 基因的表达模式惊人地匹配。转基因表达是上皮特异性的，并且在结缔组织、肌肉、骨骼、软骨、血液和内皮细胞中检测不到。值得注意的是，在气管粘膜下腺中检测到高水平的表达。总之，这些结果表明我们的 K18 表达盒在囊性纤维化患者的基因治疗中具有很高的临床应用潜力。
Targeting therapeutic gene expression to disease-affected tissues is an essential component of effective and safe gene therapy. After birth, CFTR gene expression in human lungs is localized predominantly in the epithelial cells lining the upper airways, especially in the ducts and serous tubules of the submucosal glands. We have developed a K18 expression cassette, based on the DNA control elements of the human cytokeratin 18 gene. Temporal and spatial analyses of transgenic mice demonstrated that this expression cassette targets transgene expression to almost all cell types in which CFTR is expressed. Airway epithelium expression started as early as 11.5 days of gestational age and continued into the adulthood of the transgenic mice. In these adult mice, the pattern of the reporter expression strikingly matched that of the human cytokeratin 18 and human CFTR genes. The transgene expression was epithelium-specific and undetectable in connective tissue, muscle, bone, cartilage, blood, and endothelial cells. Significantly, high levels of expression were detected in tracheal submucosal glands. Together, these results suggest that our K18 expression cassette has a high potential for clinical application in gene therapy for patients with cystic fibrosis.