Vascular Regeneration in Ischemic Hindlimb by Adeno-Associated Virus Expressing Conditionally Silenced Vascular Endothelial Growth Factor.
Vascular Regeneration in Ischemic Hindlimb by Adeno-Associated Virus Expressing Conditionally Silenced Vascular Endothelial Growth Factor.
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DOI：
10.1161/jaha.115.001815
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发表时间：
2016-05-26
影响因子：
5.4
通讯作者：
Webster KA
中科院分区：
文献类型：
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作者：
Boden J;Lassance-Soares RM;Wang H;Wei Y;Spiga MG;Adi J;Layman H;Yu H;Vazquez-Padron RI;Andreopoulos F;Webster KA
临界肢体缺血（CLI）是外周动脉疾病的极端表现，是许多患者无法满足的主要临床需求，下肢截肢是其唯一选择。经过20年的发展，治疗性血管生成已经通过肌肉内输送促血管生成蛋白、基因和干细胞进行了临床试验。疗效一般甚至没有，最大的基因治疗CLI的3期试验报告了质粒成纤维细胞生长因子的恶化趋势。在迄今为止的所有临床试验中，基因治疗都使用了表达时间有限的无管制载体。只有腺相关病毒（AAV）和慢病毒（lentivirus）等无调节扩展表达载体在临床前模型中进行了测试。我们提出了缺血（缺氧）调节条件沉默（CS） aav -人血管内皮生长因子（hVEGF）基因递送的临床前结果，在其他策略失败的情况下显示出有效性和安全性。在CLI的BALB/c小鼠模型中，我们证明了AAV - CS - hVEGF基因治疗，而不是不受调节的AAV或质粒，载体通过肌肉或动脉内给药可以挽救肢体，防止坏死和血管再生。所有载体处理均增加了毛细血管密度，但AAV - CS - hVEGF选择性地产生了有组织的纵向动脉。AAV - CS - hVEGF治疗可可逆激活血管生成和血管生成基因，包括Notch、SDF1、血管生成素和Ephrin - B2。再氧化可抑制VEGF表达并使程序失活，无明显副作用。在临床相关的小鼠CLI模型中，血管生成生长因子在缺血区域的表达限制支持后肢安全稳定的再灌注。
Critical limb ischemia (CLI) is the extreme manifestation of peripheral artery disease, a major unmet clinical need for which lower limb amputation is the only option for many patients. After 2 decades in development, therapeutic angiogenesis has been tested clinically via intramuscular delivery of proangiogenic proteins, genes, and stem cells. Efficacy has been modest to absent, and the largest phase 3 trial of gene therapy for CLI reported a worsening trend of plasmid fibroblast growth factor. In all clinical trials to date, gene therapy has used unregulated vectors with limited duration of expression. Only unregulated extended expression vectors such as adeno‐associated virus (AAV) and lentivirus have been tested in preclinical models. We present preclinical results of ischemia (hypoxia)‐regulated conditionally silenced (CS) AAV–human vascular endothelial growth factor (hVEGF) gene delivery that shows efficacy and safety in a setting where other strategies fail. In a BALB/c mouse model of CLI, we show that gene therapy with AAV‐CS‐hVEGF, but not unregulated AAV or plasmid, vectors conferred limb salvage, protection from necrosis, and vascular regeneration when delivered via intramuscular or intra‐arterial routes. All vector treatments conferred increased capillary density, but organized longitudinal arteries were selectively generated by AAV‐CS‐hVEGF. AAV‐CS‐hVEGF therapy reversibly activated angiogenic and vasculogenic genes, including Notch,SDF1, Angiopoietin, and Ephrin‐B2. Reoxygenation extinguished VEGF expression and inactivated the program with no apparent adverse side effects. Restriction of angiogenic growth factor expression to regions of ischemia supports the safe and stable reperfusion of hindlimbs in a clinically relevant murine model of CLI.