Gangrene, revascularization, and limb function improved with E-selectin/adeno-associated virus gene therapy.
Gangrene, revascularization, and limb function improved with E-selectin/adeno-associated virus gene therapy.
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DOI：
10.1016/j.jvssci.2020.10.001
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Velazquez OC
中科院分区：
文献类型：
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作者：
Quiroz HJ;Parikh PP;Lassance-Soares RM;Regueiro MM;Li Y;Shao H;Vazquez-Padron R;Percival J;Liu ZJ;Velazquez OC
残肢抢救仍需要新的治疗性血管生成概念。e -选择素是一种细胞粘附分子，对缺血组织中新生血管所需的干细胞/祖细胞的募集至关重要。我们假设在小鼠后肢缺血和坏疽模型中，用e -选择素/腺相关病毒（AAV）基因治疗启动缺血肢体组织可以增加治疗性血管生成并改善坏疽。FVB/NJ小鼠后肢肌内注射e -选择素/AAV或LacZ/AAV，结扎股动脉诱导坏疽，同时全身注射一氧化氮合成抑制剂L-NAME （l - ng -硝基精氨酸甲酯，40 mg/kg）。通过Faber后肢外观评分评估坏疽。分别采用激光多普勒灌注显像和激光共聚焦扫描显微镜对缺血足垫进行DiI灌注，评价缺血肢体再灌注率和缺血组织血管生成率。在术后第8天（POD）进行跑步机疲劳试验以确定后肢功能。与LacZ/ aav处理小鼠（n = 7）相比，E-selectin/ aav处理小鼠（n = 10）在POD 7和POD 14的Faber缺血评分均降低（P < 0.05和P < 0.01）， POD 14的激光多普勒灌注成像再灌注指标改善（P < 0.01），坏疽足部毛细血管密度增大（P < 0.001）。E-selectin/ aav处理小鼠的运动耐量也有改善（P < 0.05），相对肌肉萎缩率降低（P < 0.01）。我们推测，E-selectin/AAV基因治疗可显著促进后肢缺血坏疽小鼠后肢血管生成、再灌注和肢体功能。我们的研究结果强调了报道的新的基因治疗方法对严重肢体缺血作为未来临床研究的潜在治疗选择。在美国，尽管使用药物和手术治疗，每年仍有150万例严重肢体缺血（CLI）截肢。因此，仍然需要新的治疗性血管生成概念来挽救肢体。我们在小鼠坏疽模型（最严重的CLI形式）中使用了一种新的基因治疗方法，以证明我们的e -选择素基因治疗的有效性。像我们这样的临床前研究是为肢体受到威胁的CLI患者开发新疗法的重要一步，这些患者没有进一步的医疗或手术选择。
Novel therapeutic angiogenic concepts for critical limb ischemia are still needed for limb salvage. E-selectin, a cell-adhesion molecule, is vital for recruitment of the stem/progenitor cells necessary for neovascularization in ischemic tissues. We hypothesized that priming ischemic limb tissue with E-selectin/adeno-associated virus (AAV) gene therapy, in a murine hindlimb ischemia and gangrene model, would increase therapeutic angiogenesis and improve gangrene. FVB/NJ mice were given intramuscular hindlimb injections of either E-selectin/AAV or LacZ/AAV and then underwent induction of gangrene via femoral artery ligation and concomitant systemic injections of the nitric oxide synthesis inhibitor L-NAME (L-NG-Nitro arginine methyl ester; 40 mg/kg). Gangrene was evaluated via the Faber hindlimb appearance score. The rate of ischemic limb reperfusion and ischemic tissue angiogenesis were evaluated using laser Doppler perfusion imaging and DiI perfusion with confocal laser scanning microscopy of the ischemic footpads, respectively. The treadmill exhaustion test was performed on postoperative day (POD) 8 to determine hindlimb functionality. The E-selectin/AAV–treated mice (n = 10) had decreased Faber ischemia scores compared with those of the LacZ/AAV–treated mice (n = 7) at both PODs 7 and 14 (P < .05 and P < .01, respectively), improved laser Doppler perfusion imaging reperfusion indexes by POD 14 (P < .01), and greater gangrene footpad capillary density (P < .001). E-selectin/AAV–treated mice also had improved exercise tolerance (P < .05) and lower relative muscular atrophy (P < .01). We surmised that E-selectin/AAV gene therapy would significantly promote hindlimb angiogenesis, reperfusion, and limb functionality in mice with hindlimb ischemia and gangrene. Our findings highlight the reported novel gene therapy approach to critical limb ischemia as a potential therapeutic option for future clinical studies. In the United States, >150,000 limb amputations are performed annually for critical limb ischemia (CLI) despite the use of medical and surgical therapy. Thus, novel therapeutic angiogenic concepts for CLI are still needed for limb salvage. We used a novel gene therapy approach in a mouse model of gangrene, the most severe form of CLI, to demonstrate the efficacy of our gene therapy with E-selectin. Preclinical studies such as ours are a vital step in the development of new therapies for CLI patients with threatened limbs and no further medical or surgical options.