Pellino-1 基因治疗改善 Flk-1 杂合子小鼠的灌注并减少组织损失，但在 MAPKAP Kinase-2 敲除小鼠后肢缺血模型中失败。
Gene therapy with Pellino-1 improves perfusion and decreases tissue loss in Flk-1 heterozygous mice but fails in MAPKAP Kinase-2 knockout murine hind limb ischemia model.
机构信息
Molecular Cardiology and Angiogenesis Laboratory, Department of Surgery, University of Connecticut School of Medicine, University of Connecticut Health Center, Farmington 06030, CT, USA.
Molecular Cardiology and Angiogenesis Laboratory, Department of Surgery, University of Connecticut School of Medicine, University of Connecticut Health Center, Farmington 06030, CT, USA; Stanley J. Dudrick, Department of Surgery, Saint Mary's Hospital, Waterbury 06706, CT, USA.
出版信息
Microvasc Res. 2022 May;141:104311. doi: 10.1016/j.mvr.2022.104311. Epub 2022 Jan 7.
OBJECTIVES
In the United States, over 8.5 million people suffer from peripheral arterial disease (PAD). Previously we reported that Pellino-1(Peli1) gene therapy reduces ischemic damage in the myocardium and skin flaps in Flk-1 [Fetal Liver kinase receptor-1 (Flk-1)/ Vascular endothelial growth factor receptor-2/VEGFR2] heterozygous (Flk-1) mice. The present study compares the angiogenic response and perfusion efficiency following hind limb ischemia (HLI) in, Flk-1 and, MAPKAPKINASE2 (MK2) knockout (KO) mice to their control wild type (WT). We also demonstrated the use of Peli1 gene therapy to improve loss of function following HLI.
STUDY DESIGN AND METHODS
Femoral artery ligation (HLI) was performed in both Flk-1 and MK2 mice along with their corresponding WT. Another set of Flk-1 and MK2 were injected with either Adeno-LacZ (Ad.LacZ) or Adeno-Peli1 (Ad.Peli1) after HLI. Hind limb perfusion was assessed by laser doppler imaging at specific time points. A standardized scoring scale is used to quantify the extent of ischemia. Histology analysis performed includes capillary density, fibrosis, pro-angiogenic and anti-apoptotic proteins.
RESULTS
Flk-1 and MK2 had a slower recovery of perfusion efficiency in the ischemic limbs than controls. Both Flk-1 and MK2 KO mice showed decreased capillary density and capillary myocyte ratios with increased fibrosis than their corresponding wild types. Ad.Peli1 injected ischemic Flk-1 limb showed improved perfusion, increased capillary density, and pro-angiogenic molecules with reduced fibrosis compared to Ad.LacZ group. No significant improvement in perfusion was observed in MK2 ischemic limb after Ad. Peli1 injection.
CONCLUSION
Deletion of Flk-1 and MK2 impairs neovascularization and perfusion following HLI. Treatment with Ad. Peli1 results in increased angiogenesis and improved perfusion in Flk-1 mice but fails to rectify perfusion in MK2 KO mice. Overall, Peli1 gene therapy is a promising candidate for the treatment of PAD.
目的
在美国，超过 850 万人患有外周动脉疾病（PAD）。此前我们报道过，Pellino-1（Peli1）基因治疗可减少 Flk-1[胎肝激酶受体-1（Flk-1）/血管内皮生长因子受体-2/VEGFR2]杂合子（Flk-1）小鼠的心肌和皮瓣的缺血性损伤。本研究比较了 Flk-1 和 MAPKAPKINASE2（MK2）基因敲除（KO）小鼠后肢缺血（HLI）后的血管生成反应和灌注效率及其相应的野生型（WT）。我们还证明了使用 Peli1 基因治疗可以改善 HLI 后的功能丧失。
研究设计和方法
在 Flk-1 和 MK2 小鼠及其相应的 WT 中进行股动脉结扎（HLI）。在 HLI 后，另一组 Flk-1 和 MK2 分别注射腺病毒-LacZ（Ad.LacZ）或腺病毒-Peli1（Ad.Peli1）。在特定时间点通过激光多普勒成像评估后肢灌注。使用标准化评分量表来量化缺血的程度。进行组织学分析包括毛细血管密度、纤维化、促血管生成和抗凋亡蛋白。
结果
与对照组相比，Flk-1 和 MK2 缺血肢体的灌注效率恢复较慢。Flk-1 和 MK2 KO 小鼠的毛细血管密度和毛细血管心肌细胞比例均低于相应的野生型，纤维化程度增加。与 Ad.LacZ 组相比，注射 Ad.Peli1 的缺血 Flk-1 肢体显示出灌注改善、毛细血管密度增加和促血管生成分子减少以及纤维化减少。在注射 Ad. Peli1 后，MK2 缺血肢体的灌注没有明显改善。
结论
Flk-1 和 MK2 的缺失可损害 HLI 后的新生血管形成和灌注。Ad. Peli1 的治疗可增加 Flk-1 小鼠的血管生成并改善灌注，但不能纠正 MK2 KO 小鼠的灌注。总体而言，Peli1 基因治疗是治疗 PAD 的有前途的候选药物。
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