ELECTROPORATION-MEDIATED GENE DELIVERY OF NA+,K+-ATPASE, AND ENAC SUBUNITS TO THE LUNG ATTENUATES ACUTE RESPIRATORY DISTRESS SYNDROME IN A TWO-HIT PORCINE MODEL
ELECTROPORATION-MEDIATED GENE DELIVERY OF NA+,K+-ATPASE, AND ENAC SUBUNITS TO THE LUNG ATTENUATES ACUTE RESPIRATORY DISTRESS SYNDROME IN A TWO-HIT PORCINE MODEL
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DOI：
10.1097/shk.0000000000000228
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发表时间：
2015-01-01
期刊：
影响因子：
3.1
通讯作者：
Dean, David A.
中科院分区：
文献类型：
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作者：
Emr, Bryanna M.;Roy, Shreyas;Dean, David A.
简介：急性呼吸窘迫综合征（ARDS）是器官衰竭的常见原因，相关死亡率为40%。初始事件是肺泡-毛细血管界面的破坏，导致水肿渗漏到肺泡中。假设：电穿孔介导的上皮钠通道（ENaC）和Na+，K+-ATP酶基因导入肺泡细胞，可促进肺泡水肿的清除，减轻ARDS。研究方法：将猪麻醉并装备器械，并且夹闭上级肠系膜动脉以通过粪便凝块植入引起肠缺血/再灌注损伤和腹膜脓毒症。根据ARDSnet方案对动物进行通气。损伤后4小时，将动物随机分成以下组：（i）处理组：Na+，K+-ATP酶/ENaC质粒（n = 5）和（ii）对照组：空质粒（n = 5）。使用支气管镜将血浆递送至肺。使用八个方波电脉冲穿过胸部递送电穿孔。电穿孔后，监测猪48小时。结果：治疗组Pao（2）/Fio（2）比值及肺顺应性均高于对照组。治疗组肺湿/干比较低。在接受处理质粒的整个肺中，Na+，K+-ATP酶转基因的相对表达更高。定量组织病理学显示治疗组肺泡内纤维蛋白减少。支气管肺泡灌洗显示治疗组肺表面活性蛋白B升高。治疗组的生存率有所改善。结论：电穿孔介导的Na+，K+-ATP酶/ENaC质粒转移改善了肺功能，减少了纤维蛋白沉积，减轻了肺水肿，并提高了ARDS转化猪模型的存活率。基因治疗可以在高保真动物模型中减轻ARDS的病理生理学，为患者提供了一种潜在的新治疗方法。
Introduction: Acute respiratory distress syndrome (ARDS) is a common cause of organ failure with an associated mortality rate of 40%. The initiating event is disruption of alveolar-capillary interface causing leakage of edema into alveoli. Hypothesis: Electroporation-mediated gene delivery of epithelial sodium channel (ENaC) and Na+,K+-ATPase into alveolar cells would improve alveolar clearance of edema and attenuate ARDS. Methods: Pigs were anesthetized and instrumented, and the superior mesenteric artery was clamped to cause gut ischemia/reperfusion injury and peritoneal sepsis by fecal clot implantation. Animals were ventilated according to ARDSnet protocol. Four hours after injury, animals were randomized into groups: (i) treatment: Na+,K+-ATPase/ENaC plasmid (n = 5) and (ii) control: empty plasmid (n = 5). Plasmids were delivered to the lung using bronchoscope. Electroporation was delivered using eight-square-wave electric pulses across the chest. Following electroporation, pigs were monitored 48 h. Results: The Pao(2)/Fio(2) ratio and lung compliance were higher in the treatment group. Lung wet/dry ratio was lower in the treatment group. Relative expression of the Na+,K+-ATPase transgene was higher throughout lungs receiving treatment plasmids. Quantitative histopathology revealed a reduction in intra-alveolar fibrin in the treatment group. Bronchoalveolar lavage showed increased surfactant protein B in the treatment group. Survival was improved in the treatment group. Conclusions: Electroporation-mediated transfer of Na+,K+-ATPase/ENaC plasmids improved lung function, reduced fibrin deposits, decreased lung edema, and improved survival in a translational porcine model of ARDS. Gene therapy can attenuate ARDS pathophysiology in a high-fidelity animal model, suggesting a potential new therapy for patients.