Attenuation of Monocrotaline-Induced Pulmonary Hypertension by Luminal Adeno-Associated Virus Serotype 9 Gene Transfer of Prostacyclin Synthase
Attenuation of Monocrotaline-Induced Pulmonary Hypertension by Luminal Adeno-Associated Virus Serotype 9 Gene Transfer of Prostacyclin Synthase
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DOI：
10.1089/hum.2013.187
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发表时间：
2014-06-01
影响因子：
4.2
通讯作者：
Johnson, Larry G.
中科院分区：
文献类型：
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作者：
Gubrij, Igor B.;Martin, Sara Rebecca;Johnson, Larry G.
特发性肺动脉高压(IPAH)具有较高的发病率和死亡率。我们评价了腔内转染腺相关病毒(AAV)载体的人前列环素合成酶(HPGIS)基因能否减轻PAH。在巨细胞病毒增强型鸡肌动蛋白(CB)启动子或载体(生理盐水)的控制下，将含hPGIS基因的AAV5和AAV9载体分别注入大鼠肺内。两天后，给大鼠注射野百合碱(MCT，60 mg/kg)或生理盐水。当大鼠出现症状时(3-4周)或6周时，进行生化、血流动力学和形态学评估。与MCT对照组相比，腔内(呼吸道)注射AAV5和AAV9CBhPGIS载体(MCT-AAV5和MCT-AAV9)的大鼠血浆6-keto-PGF1水平显著升高，与未经MCT处理的大鼠(生理盐水)的水平相近。MCT对照组大鼠右室(RV)/左室(LV)+室间隔(S)比值和右室收缩压(RVSP)高于生理盐水组，而MCT-AAV5CBhPGIS和MCT-AAV9CBhPGIS大鼠的右室/左室+室间隔比值和右室收缩压与生理盐水组相似。组织切片可见MCT对照组大鼠肺小动脉肌层增厚，而MCT-AAV5CBhPGIS组和MCT-AAV9CBhPGIS组大鼠肺小动脉肌层厚度与生理盐水对照组相似。在不同启动子的实验中，用含有CB启动子的AAV9-hPGIS载体转导MCT治疗的大鼠，发现肺组织匀浆中PGF1表达水平有增加的趋势，但血浆中没有。这与MCT-AAV9CBhPGIS大鼠RV/LV+S比值和RVSP显著降低相关，与生理盐水大鼠的水平相似。在含有修饰CB启动子(Cb7)或远端上皮细胞特异性启动子(CC10)的AAV9-hPGIS载体转导的大鼠中，未检测到PGF1、RV/LV+S或RVSP水平的变化。因此，AAV9CBhPGIS载体阻止了MCT诱导的PAH的发展和相关的肺血管重塑。
Idiopathic pulmonary arterial hypertension (iPAH) is associated with high morbidity and mortality. We evaluated whether luminal delivery of the human prostacyclin synthase (hPGIS) cDNA with adeno-associated virus (AAV) vectors could attenuate PAH. AAV serotype 5 (AAV5) and AAV9 vectors containing the hPGIS cDNA under the control of a cytomegalovirus-enhanced chicken -actin (CB) promoter or vehicle (saline) were instilled into lungs of rats. Two days later, rats were injected with monocrotaline (MCT, 60mg/kg) or saline. Biochemical, hemodynamic, and morphologic assessments were performed when the rats developed symptoms (3-4 weeks) or at 6 weeks. Luminal (airway) administration of AAV5 and AAV9CBhPGIS vectors (MCT-AAV5 and MCT-AAV9 rats) significantly increased plasma levels of 6-keto-PGF1 as compared with MCT-controls, and closely resembled levels measured in rats not treated with MCT (saline-saline). Right ventricular (RV)/left ventricular (LV)+septum (S) ratios and RV systolic pressure (RVSP) were greater in MCT-control rats than in saline-saline rats, whereas the ratios and RVSP in MCT-AAV5CBhPGIS and MCT-AAV9CBhPGIS rats were similar to saline-saline rats. Thickening of the muscular media of small pulmonary arteries of MCT-control rats was detected in histological sections, whereas the thickness of the muscular media in MCT-AAV5CBhPGIS and MCT-AAV9CBhPGIS rats was similar to saline-saline controls. In experiments with different promoters, a trend toward increased levels of PGF1 expression was detected in lung homogenates, but not plasma, of MCT-treated rats transduced with an AAV9-hPGIS vector containing a CB promoter. This correlated with significant reductions in the RV/LV+S ratio and RVSP in MCT-AAV9CBhPGIS rats that resembled levels in saline-saline rats. No changes in levels of PGF1, RV/LV+S, or RVSP were detected in rats transduced with AAV9-hPGIS vectors containing a modified CB promoter (CB7) or a distal epithelial cell-specific promoter (CC10). Thus, AAV9CBhPGIS vectors prevented development of MCT-induced PAH and associated pulmonary vascular remodeling.