Augmentation of therapeutic angiogenesis using genetically modified human endothelial progenitor cells with altered glycogen synthase kinase-3β activity
Augmentation of therapeutic angiogenesis using genetically modified human endothelial progenitor cells with altered glycogen synthase kinase-3β activity
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DOI：
10.1074/jbc.m402088200
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发表时间：
2004-11-19
影响因子：
4.8
通讯作者：
Kim, HS
中科院分区：
文献类型：
--
作者：
Choi, JH;Hur, J;Kim, HS
以前我们报道过糖原合成酶激酶3 β (gsk3 β)是许多细胞内信号通路的关键调节因子，抑制糖原合成酶激酶3 β可增强血管内皮细胞的存活和迁移。在这里，我们研究了抑制gsk3 β活性对内皮祖细胞（EPC）血管生成功能的影响，并展示了一种使用转基因EPC的新的血管生成治疗策略。正如我们之前报道的那样，两种生物学上截然不同的EPC类型，纺锤形的“早期EPC”和鹅卵石形的“晚期EPC”可以从人外周血中培养出来。催化失活的gsk3 β基因被转导到两个EPC中。抑制gsk3 β信号通路导致β -连环蛋白核易位增加，血管生成细胞因子（血管内皮生长因子和白细胞介素-8）分泌增加。它促进了早期EPC的生存和增殖，促进了晚期EPC的生存和分化。与未转导的EPC相比，将这两种基因改造的EPC移植到裸小鼠缺血后肢模型中，可显著改善血流量、肢体保留和组织毛细血管密度。抑制这两种转基因EPC的gsk3 β信号传导增强了这些细胞群在体外和体内的血管生成能力。这些数据证明GSK3beta在EPC的血管生成特性中起关键作用。此外，对EPC进行基因修饰以改变这一信号传导步骤可以提高基于细胞的治疗性血管生成的功效。
Previously we reported that inhibition of glycogen synthase kinase-3beta (GSK3beta), a key regulator in many intracellular signaling pathways, enhances the survival and migration of vascular endothelial cells. Here we investigated the effect of inhibition of GSK3beta activity on the angiogenic function of endothelial progenitor cell (EPC) and demonstrated a new therapeutic angiogenesis strategy using genetically modified EPC. As we previously reported, two biologically distinct types of EPC, spindle-shaped "early EPC" and cobblestone-shaped "late EPC" could be cultivated from human peripheral blood. Catalytically inactive GSK3beta gene was transduced into both EPC. Inhibition of GSK3beta signaling pathway led to increased nuclear translocation of beta-catenin and increased secretion of angiogenic cytokines (vascular endothelial growth factor and interleukin-8). It enhanced the survival and proliferation of early EPC, whereas it promoted the survival and differentiation of late EPC. Transplantation of either of these genetically modified EPC into the ischemic hind limb model of athymic nude mouse significantly improved blood flow, limb salvage, and tissue capillary density compared with nontransduced EPC. Inhibition of GSK3beta signaling of either of these genetically modified EPC augmented the in vitro and in vivo angiogenic potency of these cell populations. These data provide evidence that GSK3beta has a key role in the angiogenic properties of EPC. Furthermore, the genetic modification of EPC to alter this signaling step can improve the efficacy of cell-based therapeutic vasculogenesis.