Cotransfection of Vascular Endothelial Growth Factor-A and Platelet-Derived Growth Factor-B Via Recombinant Adeno-Associated Virus Resolves Chronic Ischemic Malperfusion Role of Vessel Maturation
Cotransfection of Vascular Endothelial Growth Factor-A and Platelet-Derived Growth Factor-B Via Recombinant Adeno-Associated Virus Resolves Chronic Ischemic Malperfusion Role of Vessel Maturation
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DOI：
10.1016/j.jacc.2010.03.050
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发表时间：
2010-07-27
影响因子：
24
通讯作者：
Boekstegers, Peter
中科院分区：
文献类型：
--
作者：
Kupatt, Christian;Hinkel, Rabea;Boekstegers, Peter
目的：研究心肌型腺相关病毒载体（AAV 2.9 =重组腺相关病毒[rAAV]）在兔后肢缺血模型和猪冬眠心肌模型中诱导血管内皮生长因子（VEGF）-A和血小板衍生生长因子（PDGF）-B表达的能力。方法在兔股动脉切除术后第7天，将0.5 × 10（11）个rAAV.VEGF-A颗粒与或不与1 × 10（12）个rAAV.PDGF-B颗粒一起回输。在第7天和第35天，测定侧支循环计数和灌注，每个值以第35天/第7天的比率给出。在第35天测定毛细血管与肌纤维的比率。在猪中，将复位覆膜支架植入旋动脉导致第28天完全闭塞。在该时间点，反向输注rAAV。进行VEGF-A（1 × 10（13）颗粒）、rAAV.VEGF-A/PDGF-B（分别为2 × 10（12）和4 × 10（12）颗粒）或模拟转染。结果rAAV.VEGF-A能明显诱导兔血管新生，并能显著降低兔左室舒张末期压和射血分数（毛细血管与肌纤维比率; 1.67 +/- 0.09 vs. 1.32 +/- 0.11，在rAAV. LacZ处理的肢体中，p < 0.05），但不是侧支生长（125 +/- 7% vs. 106 +/-7%，p = NS）或灌注（136 +/- 12% vs. 107 +/-9%，p = NS）。在VEGF-A/PDGF-B共转染的情况下，侧支生长增加到各自第7天值的146 +/-9%，灌注增加到各自第7天值的163 +/-8%（p < 0.05）。在猪模型中，反向输注rAV。VEGF-A/PDGF-B增加局部心肌血流储备，从101 +/- 4%（rAAV模拟物）至129 +/- 8%（p < 0.05），基于侧支生长（rAAV.Mock中3.2 +/- 0.3对比rAAV.VEGF-A/PDGF-B中9.0 +/- 0.4，p < 0.05），而rAAV.VEGF-A不改变血流储备（112 ± 7%）或侧支循环计数（5.2 ± 0.7）。与rAAV. VEGF-A（37 +/-2%）不同，rAAV. VEGF-A/PDGF-B可提高射血分数（55 +/-5%vs.34 +/-3%，p < 0.05）。除新生血管外，rAAV.VEGF-A/PDGF-B还改善冬眠心肌的局部和整体心肌功能。（J Am科尔心脏病学杂志2010;56：414-22）（c）美国心脏病学会基金会2010年
Objectives We set out to investigate the ability of cardiotropic adeno-associated viral vector (AAV2.9 = recombinant adeno-associated virus [rAAV]) to induce prolonged expression of vascular endothelial growth factor (VEGF)-A and platelet-derived growth factor (PDGF)-B in a rabbit hindlimb ischemia model and a pig model of hibernating myocardium.Background Gene therapy to induce angiogenesis and arteriogenesis has produced mixed results. However, long-acting viruses, such as rAAV, as well as combined induction of angiogenesis and vessel maturation might extend the therapeutic potential.Methods In rabbits, 0.5 X 10(11) particles rAAV.VEGF-A with or without 1 X 10(12) particles rAAV.PDGF-B were retroinfused at day 7 after femoral artery excision. At days 7 and 35, collateral counts and perfusion were determined, each value given as the day 35/day 7 ratio. Capillary-to-muscle fiber ratio was determined at day 35. In pigs, implantation of a reduction stent graft into the circumflex artery led to complete occlusion at day 28. At this time point, retroinfusion of rAAV. VEGF-A (1 X 10(13) particles), rAAV.VEGF-A/PDGF-B (2 X 10(12) and 4 X 10(12) particles, respectively) or mock transfection was performed. Ejection fraction and left ventricular end-diastolic pressure were assessed at days 28 and 56.Results In rabbits, rAAV.VEGF-A strongly induced angiogenesis (capillary-to-muscle fiber ratio; 1.67 +/- 0.09 vs. 1.32 +/- 0.11 in rAAV.LacZ-treated limbs, p < 0.05), but not collateral growth (125 +/- 7% vs. 106 +/- 7%, p = NS) or perfusion (136 +/- 12% vs. 107 +/- 9%, p = NS). With VEGF-A/PDGF-B cotransfection, collateral growth increased to 146 +/- 9%, perfusion to 163 +/- 8% of the respective day 7 value (p < 0.05). In the pig model, retroinfusion of rAAV. VEGF-A/PDGF-B increased regional myocardial blood flow reserve from 101 +/- 4% (rAAV.Mock) to 129 +/- 8% (p < 0.05), based on collateral growth (3.2 +/- 0.3 in rAAV.Mock vs. 9.0 +/- 0.4 in rAAV.VEGF-A/PDGF-B, p < 0.05), whereas rAAV.VEGF-A did not alter flow reserve (112 +/- 7%) or collateral count (5.2 +/- 0.7). rAAV.VEGF-A/PDGF-B improved ejection fraction (55 +/- 5% vs. 34 +/- 3% in rAAV.Mock, p < 0.05) unlike rAAV.VEGF-A (37 +/- 2%).Conclusions Retroinfusion of rAAV.VEGF-A alone induces angiogenesis, but fails to enhance collateralization and perfusion, unless PDGF-B is cotransfected. In addition to neovascularization, rAAV.VEGF-A/PDGF-B improves regional and global myocardial function in hibernating myocardium. (J Am Coll Cardiol 2010;56:414-22) (c) 2010 by the American College of Cardiology Foundation