Transcription factors in stretch-enhanced gene delivery

描述（由申请人提供）：肺上皮细胞的循环拉伸引起各种反应，包括转染后基因转移和表达增加。这种增加的基因转移似乎是在细胞内运输的水平，可能与在拉伸的肺细胞中看到的转录因子表达和激活的增强有关。我们假设循环拉伸增加了转录因子的水平以及这些因子与质粒DNA的结合，从而促进了DNA核输入的增加。该提案将评估1)循环拉伸对细胞质中质粒转录因子复合物形成的影响，2)阻断转录因子的激活是否会减少拉伸肺上皮细胞中的基因转移，以及3)拉伸激活的转录因子是否足以在未拉伸的肺上皮细胞中赋予相同程度的拉伸增强基因传递。在这些实验完成后，将对拉伸增强基因转移的机制有新的认识，这是一种很有前途的损伤肺基因治疗技术。

DESCRIPTION (provided by applicant): Cyclic stretch in lung epithelial cells causes a variety of responses, including increased gene transfer and expression following transfection. This increased gene transfer appears to be at the level of intracellular trafficking, perhaps related to the enhanced transcription factor expression and activation seen in stretched lung cells. We hypothesize that cyclic stretch increases the levels of transcription factors and the binding of these factors to plasmid DNA, thus facilitating increased nuclear import of the DNA. This proposal will evaluate 1) the effect of cyclic stretch on the formation of plasmid-transcription factor complexes in the cytoplasm, 2) whether blocking the activation of transcription factors diminishes gene transfer in stretched lung epithelial cells, and 3) if stretch-activated transcription factors alone are sufficient to confer the same degree of stretch-enhanced gene delivery in unstretched lung epithelial cells. At the completion of these experiments, new insights will be gained into the mechanism of stretch-enhanced gene transfer, a promising technique for gene therapy in the injured lung.