Barnett Rebecca C, Lin Xin, Barravecchia Michael, Norman Rosemary A, de Mesy Bentley Karen L, Fazal Fabeha, Young Jennifer L, Dean David A
1 Division of Neonatology, University of Rochester, Rochester, New York, NY 14642, USA.
2 Department of Pathology and Laboratory Medicine, University of Rochester, Rochester, New York, NY 14642, USA.
Exp Biol Med (Maywood). 2017 Jul;242(13):1345-1354. doi: 10.1177/1535370217713000. Epub 2017 Jun 5.
Surfactant Protein B Deficiency is a rare but lethal monogenetic, congenital lung disease of the neonate that is unresponsive to any treatment except lung transplantation. Based on the potential that gene therapy offers to treat such intractable diseases, our objective was to test whether an electroporation-based gene delivery approach could restore surfactant protein B expression and improve survival in a compound knockout mouse model of surfactant protein B deficiency. Surfactant protein B expression can be shut off in these mice upon withdrawl of doxycycline, resulting in decreased levels of surfactant protein B within four days and death due to lung dysfunction within four to seven days. Control or one of several different human surfactant protein B-expressing plasmids was delivered to the lung by aspiration and electroporation at the time of doxycycline removal or four days later. Plasmids expressing human surfactant protein B from either the UbC or CMV promoter expressed surfactant protein B in these transgenic mice at times when endogenous surfactant protein B expression was silenced. Mean survival was increased 2- to 5-fold following treatment with the UbC or CMV promoter-driven plasmids, respectively. Histology of all surfactant protein B treated groups exhibited fewer neutrophils and less alveolar wall thickening compared to the control groups, and electron microscopy revealed that gene transfer of surfactant protein B resulted in lamellar bodies that were similar in the presence of electron-dense, concentric material to those in surfactant protein B-expressing mice. Taken together, our results show that electroporation-mediated gene delivery of surfactant protein B-expressing plasmids improves survival, lung function, and lung histology in a mouse model of surfactant protein B deficiency and suggest that this may be a useful approach for the treatment of this otherwise deadly disease. Impact statement Surfactant protein B (SP-B) deficiency is a rare but lethal genetic disease of neonates that results in severe respiratory distress with no available treatments other than lung transplantation. The present study describes a novel treatment for this disease by transferring the SP-B gene to the lungs using electric fields in a mouse model. The procedure is safe and results in enough expression of exogenous SP-B to improve lung histology, lamellar body structure, and survival. If extended to humans, this approach could be used to bridge the time between diagnosis and lung transplantation and could greatly increase the likelihood of affected neonates surviving to transplantation and beyond.
表面活性蛋白B缺乏症是一种罕见但致命的新生儿单基因先天性肺部疾病,除肺移植外,对任何治疗均无反应。基于基因治疗在治疗此类难治性疾病方面的潜力,我们的目标是测试基于电穿孔的基因递送方法是否能在表面活性蛋白B缺乏的复合敲除小鼠模型中恢复表面活性蛋白B的表达并提高生存率。在这些小鼠中,停用强力霉素后表面活性蛋白B的表达可被关闭,导致四天内表面活性蛋白B水平下降,并在四至七天内由于肺功能障碍而死亡。在停用强力霉素时或四天后,通过吸入和电穿孔将对照质粒或几种不同的表达人表面活性蛋白B的质粒之一递送至肺部。在内源性表面活性蛋白B表达沉默时,来自UbC或CMV启动子的表达人表面活性蛋白B的质粒在这些转基因小鼠中表达表面活性蛋白B。分别用UbC或CMV启动子驱动的质粒处理后,平均生存期延长了2至5倍。与对照组相比,所有表面活性蛋白B处理组的组织学显示中性粒细胞减少,肺泡壁增厚减轻,电子显微镜显示表面活性蛋白B的基因转移导致板层小体,其电子致密的同心物质与表达表面活性蛋白B的小鼠中的相似。综上所述,我们的结果表明,电穿孔介导的表达表面活性蛋白B的质粒的基因递送可提高表面活性蛋白B缺乏小鼠模型的生存率、肺功能和肺组织学,并表明这可能是治疗这种致命疾病的一种有用方法。影响声明表面活性蛋白B(SP-B)缺乏症是一种罕见但致命的新生儿遗传性疾病,导致严重的呼吸窘迫,除肺移植外无其他可用治疗方法。本研究描述了一种通过在小鼠模型中使用电场将SP-B基因转移到肺部来治疗这种疾病的新方法。该程序是安全的,可导致外源SP-B充分表达,以改善肺组织学、板层小体结构和生存率。如果扩展到人类,这种方法可用于在诊断和肺移植之间的时间段内提供过渡,并且可以大大增加受影响新生儿存活至移植及以后的可能性。