Ibuprofen-Caffeine Modulation of Retinal Endothelial Tip Cell Migration

早产儿视网膜病变（ROP）困扰着所有极低胎龄新生儿（ELGAN，<28周/<1250 g）的多达50%。在ELGAN和动物模型中，咖啡因和咖啡因已被证明可降低严重ROP的风险。使用我们实验室开发的独特技术，我们将研究对人类视网膜微血管内皮尖端细胞（EC）的特定影响，这是异常血管生成背后的驱动力。我们将研究它们与星形胶质细胞的动态相互作用，它们在氧化应激过程中的选择，激活和迁移。更重要的是，我们将研究布洛芬和/或咖啡因在防止其激活和降低其感知血管生成线索的能力方面的功效。该提案的首要目标是研究EC尖端细胞的行为及其与星形胶质细胞在氧化应激（高氧/缺氧循环）环境中的关系;并确定布洛芬与咖啡因联合给药是否会保持FIP细胞静止。利用最先进的生物分析学、蛋白质组学、药物基因组学、生物信息学和成像技术，我们将使用三个相互关联的特定目标：1）检查正常氧和氧化应激（短暂的高氧/缺氧循环）下人视网膜微血管EC和人脑星形胶质细胞之间的关系。我们将重点关注VEGF和ECM蛋白水解、VEGF释放和梯度增加;尖端细胞活化、VEGF的释放和重新捕获以及迁移;以及VEGF和Notch信号传导机制; 2）使用小干扰RNA建立VEGFR-2、VEGFR-3、NP-1、Notch 1、D1 I4和Jagged 1在尖端细胞选择、活化和迁移中的作用 在人视网膜微血管内皮细胞中这些特异性基因的siRNA敲低。我们将研究星形胶质细胞的影响;和3）确定布洛芬与咖啡因一起增强是否会保护和保存正常人视网膜微血管EC和星形胶质细胞在氧化应激中的生长和功能。我们将使用最先进的技术来提供对生物分子机制，药代动力学，药物相互作用，药物转运和代谢的见解。

Retinopathy of prematurity (ROP) is afflicts as much as 50% of all extremely low gestational age neonates (ELGANs, <28 weeks/<1250g). Ibuprofen and caffeine have been shown to decrease the risk of severe ROP in ELGANs and in animal models. Using unique techniques developed in our laboratory, we will examine the specific effects on human retinal microvascular endothelial tip cells (ECs), the driving force behind aberrant angiogenesis. We will study their dynamic interaction with astrocytes, their selection, activation, and migration during oxidative stress. More importantly, we will examine the efficacy of ibuprofen and/or caffeine in preventing their activation and reducing their capacity to sense angiogenic cues. The overarching goal of this proposal is to study the behavior of EC tip cells and their relationship with astrocytes in the setting of oxidative stress (hyperoxia/hypoxia cycling); and to determine whether ibuprofen coadministered with caffeine will preserve fip cell quiescence. Using state-of-the-art bioanalytics, proteomics, pharmacogenomics, bioinformatics, and imaging techniques, we will use three interrelated specific aims: 1) to examine the relationship between human retinal microvascular ECs and human brain astrocytes in normoxia and in oxidative stress (brief hyperoxia/hypoxia cycling). We will focus on VEGF and ECM proteolysis, VEGF release and increased gradient; tip cell activation, release and recapture of VEGF, and migration; and VEGF and Notch signaling mechanisms; 2) to establish the roles of VEGFR-2, VEGFR-3, NP-1, Notch 1, D1I4, and Jagged 1 on tip cell selection, activation and migration using small interference RNA (SiRNA) knockdown of these specific genes in human retinal microvascular ECs. We will study the influence of astrocytes; and 3) to determine whether ibuprofen potentiated with caffeine will protect and preserve normal human retinal microvascular EC and astrocyte growth and function in oxidative stress. We will use state-of-the-art technologies to provide insights on the biomolecular mechanisms, pharmacokinetics, drug interactions; drug transport and metabolism.