In vivo imaging of gene transfer to the respiratory tract
In vivo imaging of gene transfer to the respiratory tract
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DOI：
10.1016/j.biomaterials.2007.11.017
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发表时间：
2008-04-01
期刊：
影响因子：
14
通讯作者：
Alton, Eric W. F. W.
中科院分区：
文献类型：
--
作者：
Griesenbach, Uta;Meng, Cuixiang;Alton, Eric W. F. W.
利用荧光素酶在不同器官中的表达来成像体内基因表达已经使用了好几年。与其他器官不同，肺的体内成像，特别是在非病毒基因转移后，尚未得到广泛研究。本研究的目的是解决几个问题：(1)体内发光是否以剂量依赖的方式与标准组织匀浆荧光素酶检测相关？重组仙台病毒（SeV）非常有效地转导气道上皮细胞，并用于解决这一问题。(2)检测非病毒基因转移的灵敏度是否足够？我们使用我们的标准“嗅探”方案用SeV-Lux载体治疗小鼠，这种方法主要导致肺部沉积。所有动物体内可见剂量相关的光发射。重要的是，在左肺和右肺的体内和离体检测之间存在显著相关性（r > 0.90, p < 0.0001）。接下来，我们用荧光素酶质粒（pCIKLux）与阳离子脂质GL67复合物（n = 25-27/组）通过鼻腔灌注或肺部（“嗅”）转染小鼠鼻上皮，并在转染24小时后在体内成像荧光素酶的表达。在两个器官中均可检测到基因表达。在体内和离体检测在左肺有显著相关性（r = 0.52, p < 0.005），而在右肺无显著相关性。鼻部的相关性较弱（r = 0.45, p < 0.05）。据我们所知，这些研究首次表明，这种评估肺基因转移的非侵入性方法对于评估非病毒基因转移剂是可行的。(c) 2007 Elsevier Ltd.版权所有。
Imaging of in vivo gene expression using luciferase expression in various organs has been used for several years. In contrast to other organs, in vivo imaging of the lung, particularly after non-viral gene transfer has not been extensively studied. The aim of this study was to address several questions: (1) Does in vivo light emission correlate with standard tissue homogenate-based luciferase detection in a dose-dependent manner? Recombinant Sendai virus (SeV) transduces air-way epithelial cells very efficiently and was used to address this question, (2) Is the sensitivity of the assay sufficient to detect non-viral gene transfer? We treated mice with SeV-Lux vector using our standard "sniffing" protocol, a method that predominantly results in lung deposition. Dose-related in vivo light emission was visible in all animals. Importantly, there was a significant correlation (r > 0.90, p < 0.0001) between the in vivo and ex vivo assays in both the left and right lung. We next transfected the nasal epithelium via nasal perfusion or the lungs ("sniffing") of mice with a luciferase plasmid (pCIKLux) complexed to the cationic lipid GL67 (n = 25-27/group) and imaged luciferase expression in vivo 24 h after transfection. Gene expression was detectable in both organs. Correlation between the in vivo and ex vivo assays was significant (r = 0.52, p < 0.005) in the left, but not the right lung. The correlation in the nose was weaker (r = 0.45, p < 0.05). To our knowledge these studies show for the first time that this non-invasive method of assessing pulmonary gene transfer is viable for evaluating non-viral gene transfer agents. (c) 2007 Elsevier Ltd. All rights reserved.