Lee L Y, Patel S R, Hackett N R, Mack C A, Polce D R, El-Sawy T, Hachamovitch R, Zanzonico P, Sanborn T A, Parikh M, Isom O W, Crystal R G, Rosengart T K
Department of Cardiothoracic Surgery, The New York Hospital-Cornell Medical Center, New York, USA.
Ann Thorac Surg. 2000 Jan;69(1):14-23; discussion 23-4. doi: 10.1016/s0003-4975(99)01102-9.
Adenovirus (Ad) vector-mediated gene therapy strategies have emerged as promising modalities for the "biological revascularization" of tissues. We hypothesized that direct intramyocardial, as opposed to intracoronary, administration of an Ad vector coding for the vascular endothelial growth factor 121 cDNA (Ad(GV)VEGF121.10) would provide highly focal Ad genome levels, and increases in VEGF, ideal for inducing localized therapeutic angiogenesis.
Persistence and regional distribution of the vector were assessed by TaqMan real-time quantitative polymerase chain reaction technology and enzyme-linked immunosorbent assay, after intramyocardial Ad(GV)VEGF121.10 in the rat, and either intramyocardial or intracoronary (circumflex territory) vector in Yorkshire swine. Based on these results, we assessed the focal nature of the improved cardiac blood flow in a previously reported porcine myocardial ischemia model.
Intramyocardial delivery of Ad(GV)VEGF121.10 in the rat resulted in local persistence of the Ad genome that decreased 1,000-fold over 3 weeks, with peak myocardial VEGF expression 24 to 72 h after vector delivery. After intramyocardial Ad(GV)VEGF121.10 in the circumflex distribution of pigs, Ad vector genome and VEGF protein levels were more than 1,000-fold and more than 90-fold higher, respectively, in this distribution than in other myocardial regions. In comparison, intracoronary injection yielded maximum myocardial Ad genome and VEGF levels 33-fold and 9-fold lower, respectively, than that after intramyocardial delivery. Angiograms obtained 28 days after intramyocardial Ad(GV)VEGF121.10 demonstrated rapid circumflex reconstitution via collaterals localized to the region of vector administration.
These studies demonstrate that direct intramyocardial administration of Ad(GV)VEGF121.10 results in focal genome and VEGF levels, including focal angiogenesis, sufficient to normalize blood flow to the ischemic myocardium, findings that are relevant to designing human trials of gene therapy-mediated cardiac angiogenesis.
腺病毒（Ad）载体介导的基因治疗策略已成为组织“生物血管再生”的有前景的方式。我们假设，与冠状动脉内给药相反，直接心肌内注射编码血管内皮生长因子121 cDNA的腺病毒载体（Ad(GV)VEGF⒈⒈）可提供高度局灶性的腺病毒基因组水平，并使血管内皮生长因子（VEGF）增加，这对于诱导局部治疗性血管生成是理想的。
在大鼠心肌内注射Ad(GV)VEGF⒈⒈后，以及在约克郡猪心肌内或冠状动脉内（回旋支区域）注射载体后，通过TaqMan实时定量聚合酶链反应技术和酶联免疫吸附测定法评估载体的持久性和区域分布。基于这些结果，我们在先前报道的猪心肌缺血模型中评估了改善的心脏血流的局灶性。
在大鼠心肌内注射Ad(GV)VEGF⒈⒈导致腺病毒基因组在局部持续存在，在3周内下降1000倍，载体注射后24至72小时心肌VEGF表达达到峰值。在猪的回旋支分布区域进行心肌内注射Ad(GV)VEGF⒈⒈后，该分布区域的腺病毒载体基因组和VEGF蛋白水平分别比其他心肌区域高1000倍以上和90倍以上。相比之下，冠状动脉内注射产生的最大心肌腺病毒基因组和VEGF水平分别比心肌内注射后低33倍和9倍。心肌内注射Ad(GV)VEGF⒈⒈28天后获得的血管造影显示，通过局限于载体给药区域的侧支循环，回旋支迅速重建。
这些研究表明，直接心肌内注射Ad(GV)VEGF⒈⒈可导致局灶性基因组和VEGF水平，包括局灶性血管生成，足以使缺血心肌的血流正常化，这些发现与设计基因治疗介导的心脏血管生成的人体试验相关。
