Mesenchymal stem cell-based gene therapy with prostacyclin synthase enhanced neovascularization in hindlimb ischemia
Mesenchymal stem cell-based gene therapy with prostacyclin synthase enhanced neovascularization in hindlimb ischemia
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DOI：
10.1016/j.atherosclerosis.2009.02.023
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发表时间：
2009-09-01
期刊：
影响因子：
5.3
通讯作者：
Murohara, Toyoaki
中科院分区：
文献类型：
--
作者：
Ishii, Masakazu;Numaguchi, Yasushi;Murohara, Toyoaki
目的：骨髓细胞疗法通过祖细胞和肌细胞本身分泌血管生成因子促进侧支形成，从而为严重肢体缺血患者提供了一种新的选择。然而，一些病例由于移植物失败而对这种疗法难治。因此，我们使用遗传修饰的间充质干细胞（MSC）过表达血管调节蛋白，前列环素（PGI（2）），以检查它是否可以提高移植和新血管形成在后肢ischemia.Methods和结果：我们设计的过度表达PGI（2）合成酶（PGIS）在MSC，这导致更高的磷酸化Akt和Bcl-2的表达水平比对照组。在缺氧的条件下。PGIS的过表达导致环氧化酶-2和过氧化物酶体增殖物激活受体8的表达上调，随后MSC的增殖率增加40%。然后，我们在C57 BL 6/J小鼠中产生单侧后肢缺血，所述小鼠注射用GFP转染的MSC、过表达PGIS的MSC或载体。激光多普勒分析表明，MSC的管理有效地恢复血液灌注，并在手术后7天内达到峰值血流量与MSC过表达PGIS的小鼠，这是早于与GFP转染的MSC小鼠。结论：持续释放PGI（2）可增强MSCs的促血管生成功能，促进缺血组织中肌细胞的再生，提示细胞基因治疗对严重肢体缺血具有潜在的治疗价值。(C)2009爱思唯尔爱尔兰有限公司保留所有权利。
Objective: Bone marrow cell therapy contributes to collateral formation through the secretion of angiogenic factors by progenitor cells and muscle cells per se, thereby presenting a novel option for patients with critical limb ischemia. However, some cases are refractory to this therapy due to graft failure. Therefore, we used genetic modification of mesenchymal stem cells (MSCs) to overexpress a vasoregulatory protein, prostacyclin (PGI(2)), to examine whether it could enhance engraftment and neovascularization in hindlimb ischemia.Methods and results: We engineered the overexpression of PGI(2) synthase (PGIS) within MSCs, which resulted in higher expression levels of phosphorylated Akt and Bcl-2 than in control. Under hypoxic conditions. the overexpression of PGIS led to upregulated expression of cyclooxigenase-2 and peroxisome proliferator-activated receptor 8, following a 40% increased rate of proliferation in MSCs. We then produced unilateral hindlimb ischemia in C57BL6/J mice, which were injected either with MSCs transfected with GFP, with MSCs overexpressing PGIS, or with vehicle. Laser Doppler analyses demonstrated that the administration of MSCs effectively recovered blood perfusion, and that the peak blood flow was reached within 7 days of surgery in mice with MSCs overexpressing PGIS, which was earlier than that in mice with MSCs transfected with GFP. This beneficial effect was correlated to enhanced collateral formation and muscle bundle proliferation.Conclusion: Sustained release of PGI(2) enhanced the proangiogenic function of MSCs and subsequent muscle cell regrowth in the ischemic tissue suggesting potential therapeutic benefits of cell-based gene therapy for critical limb ischemia. (C) 2009 Elsevier Ireland Ltd. All rights reserved.