Inhalation of Nebulized Perfluorochemical Enhances Recombinant Adenovirus and Adeno-Associated Virus-Mediated Gene Expression in Lung Epithelium
Inhalation of Nebulized Perfluorochemical Enhances Recombinant Adenovirus and Adeno-Associated Virus-Mediated Gene Expression in Lung Epithelium
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DOI：
10.1089/hgtb.2012.014
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发表时间：
2012-04-01
影响因子：
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通讯作者：
Weiss, Daniel J.
中科院分区：
文献类型：
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作者：
Beckett, Travis;Bonneau, Laura;Weiss, Daniel J.
在肺内载体给药期间使用全氟化学液体增强重组腺病毒和腺相关病毒（AAV）介导的肺上皮基因表达。我们假设，吸入雾化全氟化学蒸气也会增强上皮细胞基因表达后，随后的intrichiheal载体管理。将自由呼吸的成年C57 BL/6小鼠暴露于密封的有机玻璃室中的雾化全氟溴龙或无菌盐水中选定的时间。重组腺病毒载体在之后的选定时间通过经气管穿刺施用，并且在载体施用后3天杀死小鼠以评估转基因表达。小鼠耐受雾化吸入全氟龙，无明显不良反应。与单独使用载体或盐水加载体对照相比，在雾化全氟纶暴露后6小时施用载体导致气道和肺泡上皮中基因表达平均增加540%（p < 0.05）。然而，在全氟龙暴露后1小时、1天或3天给予载体与雾化盐水与载体或单独载体没有区别，需要暴露于雾化全氟龙60分钟。在猕猴的平行初步研究中，吸入雾化的perflubron增强了重组AAV 2/5载体在整个肺中的表达。连续胸部X线片、支气管肺泡灌洗、全血细胞计数和血清生化结果表明，雾化吸入perflubron无明显不良反应。此外，对仅接受全氟溴铵雾化的一只猕猴进行了1年的监测，未发现明显的暴露不良反应。这些结果表明，吸入雾化全氟溴铵，一个简单的，临床上更可行的技术比液体全氟溴铵气管内给药，安全地增强肺基因表达。
Use of perfluorochemical liquids during intratracheal vector administration enhances recombinant adenovirus and adeno-associated virus (AAV)-mediated lung epithelial gene expression. We hypothesized that inhalation of nebulized perfluorochemical vapor would also enhance epithelial gene expression after subsequent intratracheal vector administration. Freely breathing adult C57BL/6 mice were exposed for selected times to nebulized perflubron or sterile saline in a sealed Plexiglas chamber. Recombinant adenoviral vector was administered by transtracheal puncture at selected times afterward and mice were killed 3 days after vector administration to assess transgene expression. Mice tolerated the nebulized perflubron without obvious ill effects. Vector administration 6 hr after nebulized perflubron exposure resulted in an average 540% increase in gene expression in airway and alveolar epithelium, compared with that with vector alone or saline plus vector control (p < 0.05). However, vector administration 1 hr, 1 day, or 3 days after perflubron exposure was not different from either nebulized saline with vector or vector alone and a 60-min exposure to nebulized perflubron is required. In parallel pilot studies in macaques, inhalation of nebulized perflubron enhanced recombinant AAV2/5 vector expression throughout the lung. Serial chest radiographs, bronchoalveolar lavages, and results of complete blood counts and serum biochemistries demonstrated no obvious adverse effects of nebulized perflubron. Further, one macaque receiving nebulized perflubron only was monitored for 1 year with no obvious adverse effects of exposure. These results demonstrate that inhalation of nebulized perflubron, a simple, clinically more feasible technique than intratracheal administration of liquid perflubron, safely enhances lung gene expression.