Department of Cardiovascular Medicine, Kyoto Prefectural University School of Medicine, Kyoto 602-8566, Japan.
Proc Natl Acad Sci U S A. 2011 Jun 7;108(23):9472-7. doi: 10.1073/pnas.1101296108. Epub 2011 May 18.
Endothelial and endothelial progenitor cells (ECs and EPCs) play a fundamental role in angiogenesis that is essential for numerous physiological and pathological processes. The phosphatase and tensin homolog (PTEN)/ phosphoinositide 3-kinase (PI3K) pathway has been implicated in angiogenesis, but the mechanism in the regulation of this pathway in ECs and EPCs is poorly understood. Here we show that ARIA (apoptosis regulator through modulating IAP expression), a transmembrane protein that we recently identified, regulates the PTEN/PI3K pathway in ECs and EPCs and controls developmental and postnatal angiogenesis in vivo. We found that ARIA is abundantly expressed in EPCs and regulates their angiogenic functions by modulating PI3K/Akt/endothelial nitric oxide synthase (eNOS) signaling. Genetic deletion of ARIA caused nonfatal bleeding during embryogenesis, in association with increased small vessel density and altered expression of various vascular growth factors including angiopoietins and VEGF receptors. Postnatal neovascularization induced by critical limb ischemia was substantially enhanced in ARIA-null mice, in conjunction with more bone marrow (BM)-derived ECs detected in ischemic muscles. Administration of PI3K or NO synthase inhibitor completely abolished the enhanced neovascularization in ARIA(-/-) mice. Mechanistically, we identified that ARIA interacts with PTEN at the intracellular domain independently of the PTEN phosphorylation in its C-terminal tail. Overexpressed ARIA increased PTEN in the membrane fraction, whereas ARIA-silencing reduced the membrane-associated PTEN, resulting in modified PI3K/Akt signaling. Taken together, our findings establish a previously undescribed mode of regulation of the PTEN/PI3K/Akt pathway by ARIA, and reveal a unique mechanism in the control of angiogenesis. These functions of ARIA might offer a unique therapeutic potential.
内皮细胞和内皮祖细胞(EC 和 EPC)在血管生成中发挥着基本作用,血管生成对于许多生理和病理过程都是必不可少的。磷酸酶和张力蛋白同源物(PTEN)/ 磷脂酰肌醇 3-激酶(PI3K)途径已被牵涉到血管生成中,但其在 EC 和 EPC 中调节该途径的机制尚不清楚。在这里,我们表明,我们最近鉴定的跨膜蛋白 ARIA(通过调节 IAP 表达来调节细胞凋亡),调节 EC 和 EPC 中的 PTEN/PI3K 途径,并控制体内的发育和出生后血管生成。我们发现,ARIA 在 EPC 中大量表达,并通过调节 PI3K/Akt/内皮型一氧化氮合酶(eNOS)信号来调节其血管生成功能。ARIA 的基因缺失导致胚胎发生期间非致命性出血,伴随着小血管密度增加和各种血管生长因子(包括血管生成素和 VEGF 受体)的表达改变。在 ARIA 缺失小鼠中,通过严重肢体缺血诱导的新生血管化显著增强,同时在缺血肌肉中检测到更多的骨髓(BM)衍生的 EC。PI3K 或一氧化氮合酶抑制剂的给药完全消除了 ARIA(-/-)小鼠中增强的新生血管化。在机制上,我们确定 ARIA 与 PTEN 在其 C 末端尾部的 PTEN 磷酸化独立的细胞内结构域相互作用。过表达的 ARIA 增加了膜部分中的 PTEN,而 ARIA 沉默减少了膜相关的 PTEN,导致修改后的 PI3K/Akt 信号。总之,我们的研究结果确立了 ARIA 对 PTEN/PI3K/Akt 途径的一种以前未描述的调节模式,并揭示了血管生成控制的独特机制。ARIA 的这些功能可能提供独特的治疗潜力。