Gubrij Igor B, Martin Sara Rebecca, Pangle Amanda K, Kurten Richard, Johnson Larry G
1 Division of Pulmonary and Critical Care, Department of Medicine, University of Arkansas for Medical Sciences , Little Rock, AR 72205.
Hum Gene Ther. 2014 Jun;25(6):498-505. doi: 10.1089/hum.2013.187. Epub 2014 Mar 26.
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 β-actin (CB) promoter or vehicle (saline) were instilled into lungs of rats. Two days later, rats were injected with monocrotaline (MCT, 60 mg/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(α) 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(α) 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(α), 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.
特发性肺动脉高压（iPAH）与高发病率和死亡率相关。我们评估了用腺相关病毒（AAV）载体经腔递送人前列环素合酶（hPGIS）cDNA是否能减轻肺动脉高压。将含有在巨细胞病毒增强的鸡β-肌动蛋白（CB）启动子控制下的hPGIS cDNA的AAV血清型5（AAV5）和AAV9载体或载体（生理盐水）注入大鼠肺中。两天后，给大鼠注射野百合碱（MCT，60mg/kg）或生理盐水。当大鼠出现症状时（3 - 4周）或在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，经腔（气道）给予AAV5和AAV9CBhPGIS载体（MCT - AAV5和MCT - AAV9大鼠）显著提高了血浆6 - 酮 - PGF1（α）水平，且与未用MCT处理的大鼠（生理盐水 - 生理盐水组）测得的水平相近。MCT对照组大鼠的右心室（RV）/左心室（LV）+室间隔（S）比值和RV收缩压（RVSP）高于生理盐水 - 生理盐水组大鼠，而MCT - AAV5CBhPGIS和MCT - AAV9CBhPGIS大鼠的比值和RVSP与生理盐水 - 生理盐水组大鼠相似。在组织学切片中检测到MCT对照组大鼠小肺动脉肌层增厚，而MCT - AAV5CBhPGIS和MCT - AAV9CBhPGIS大鼠的肌层厚度与生理盐水 - 生理盐水对照组相似。在不同启动子的实验中，在用含有CB启动子的AAV9 - hPGIS载体转导的MCT处理大鼠的肺匀浆中检测到PGF1（α）表达水平有升高趋势，但血浆中未检测到。这与MCT - AAV9CBhPGIS大鼠的RV/LV + S比值和RVSP显著降低相关，其水平与生理盐水 - 生理盐水组大鼠相似。在用含有修饰的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（α）水平、RV/LV + S或RVSP的变化。因此，AAV9CBhPGIS载体可预防MCT诱导的PAH及相关肺血管重塑的发生。
