mTORC1 and mTORC2 Play Different Roles in the Functional Survival of Transplanted Adipose-Derived Stromal Cells in Hind Limb Ischemic Mice Via Regulating Inflammation In Vivo
mTORC1 and mTORC2 Play Different Roles in the Functional Survival of Transplanted Adipose-Derived Stromal Cells in Hind Limb Ischemic Mice Via Regulating Inflammation In Vivo
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mTORC1 和 mTORC2 通过调节体内炎症在后肢缺血小鼠移植的脂肪源性基质细胞的功能存活中发挥不同的作用
DOI：
10.1002/stem.1265
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发表时间：
2013-01-01
期刊：
影响因子：
5.2
通讯作者：
Cao, Feng
中科院分区：
文献类型：
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作者：
Fan, Weiwei;Cheng, Kang;Cao, Feng
细胞存活率低严重限制了外周动脉疾病（PAD）干细胞治疗的有益效果。本研究旨在研究哺乳动物雷帕霉素靶蛋白（mTOR）在小鼠PAD模型中移植的小鼠脂肪源性基质细胞（mADSC）的存活和治疗功能中的作用。从双报告萤火虫荧光素酶和增强型绿色荧光蛋白阳性转基因小鼠中分离mADSC（1.0 × 10（7）），在股动脉结扎/切除后肌内植入C57 BL/6小鼠的后肢，并使用非侵入性生物发光成像（BLI）进行监测。尽管移植的mADSC通过调节涉及mTOR通路的炎症和血管生成细胞因子反应在体内产生抗凋亡/促血管生成作用，但纵向BLI显示缺血后肢中移植后mADSC在类似4周内进行性死亡。使用低剂量雷帕霉素治疗mADSC选择性靶向mTOR复合物-1（mTORC 1）减弱了促炎细胞因子（白细胞介素[IL]-1 β和肿瘤坏死因子-α [TNF-α]）表达和中性粒细胞/巨噬细胞浸润，这明显促进了体内mADSC的活力和抗凋亡/促血管生成功效。然而，使用PP 242或高剂量雷帕霉素靶向双重mTORC 1/mTORC 2导致IL-1 β/TNF-α上调和抗炎性IL-10、IL-6和血管内皮生长因子/血管内皮生长因子受体2下调，破坏了体内mADSC的存活和抗凋亡/促血管生成作用。此外，低剂量雷帕霉素废除mADSC的TNF-α分泌，并在体外将细胞从缺氧/复氧诱导的死亡中拯救出来，而PP 242或高剂量雷帕霉素发挥促炎作用并促进细胞死亡。总之，mTORC 1和mTORC 2可能差异调节炎症并影响移植的mADSC在缺血后肢中的功能存活。这些发现揭示了mTOR可能演变为机制驱动方法的有希望的候选者，以促进基于细胞的PAD治疗的翻译。干细胞2013;31：203-214
Poor cell survival severely limits the beneficial effects of stem cell therapy for peripheral arterial disease (PAD). This study was designed to investigate the role of mammalian target of rapamycin (mTOR) in the survival and therapeutic function of transplanted murine adipose-derived stromal cells (mADSCs) in a murine PAD model. mADSCs (1.0 x 10(7)) were isolated from dual-reporter firefly luciferase and enhanced green fluorescent protein-positive transgenic mice, intramuscularly implanted into the hind limb of C57BL/6 mice after femoral artery ligation/excision, and monitored using noninvasive bioluminescence imaging (BLI). Although engrafted mADSCs produced antiapoptotic/proangiogenic effects in vivo by modulating the inflammatory and angiogenic cytokine response involving the mTOR pathway, longitudinal BLI revealed progressive death of post-transplant mADSCs within similar to 4 weeks in the ischemic hind limb. Selectively targeting mTOR complex-1 (mTORC1) using low-dose rapamycin treatment with mADSCs attenuated proinflammatory cytokines (interleukin [IL]-1 beta and tumor necrosis factor-alpha [TNF-alpha]) expression and neutrophil/macrophage infiltration, which overtly promoted mADSCs viability and antiapoptotic/proangiogenic efficacy in vivo. However, targeting dual mTORC1/mTORC2 using PP242 or high-dose rapamycin caused IL-1 beta/TNF-alpha upregulation and anti-inflammatory IL-10, IL-6, and vascular endothelial growth factor/vascular endothelial growth factor receptor 2 downregulation, undermining the survival and antiapoptotic/proangiogenic action of mADSCs in vivo. Furthermore, low-dose rapamycin abrogated TNF-alpha secretion by mADSCs and rescued the cells from hypoxia/reoxygenation-induced death in vitro, while PP242 or high-dose rapamycin exerted proinflammatory effects and promoted cell death. In conclusion, mTORC1 and mTORC2 may differentially regulate inflammation and affect transplanted mADSCs' functional survival in ischemic hind limb. These findings uncover that mTOR may evolve into a promising candidate for mechanism-driven approaches to facilitate the translation of cell-based PAD therapy. STEM CELLS 2013;31:203-214