Hughes David, Bertelli Pietro M, Pedrini Edoardo, Allen Lynsey-Dawn, Branco Cristina, Curtis Tim M, Medina Reinhold J, Stitt Alan W, Lechner Judith
Wellcome-Wolfson Institute for Experimental Medicine, Queen's University Belfast, UK.
The Patrick G Johnston Centre for Cancer Research and Cell Biology, Queen's University Belfast, UK.
Exp Eye Res. 2025 Oct;259:110535. doi: 10.1016/j.exer.2025.110535. Epub 2025 Jul 21.
In many retinopathies, hypoxia stimulates pathogenic neovascularization. The precise impact of hypoxia and how it may drive a switch in retinal microvascular endothelial cell metabolism during active angiogenesis remains ill-defined. This study has sought to understand dynamic shifts in the metabolic profile of retinal microvascular endothelial cells exposed to hypoxia and during ischemia-induced neovascularization. The impact of manipulating glycolytic metabolism by intravitreal injection of the glycolytic inhibitor 3-(3-Pyridinyl)-1-(4pyridinyl)-2-propen-1-one) (3PO) on pre-retinal neovascularization was investigated using the oxygen-induced retinopathy (OIR) model. Metabolic pathways generating adenosine triphosphate (ATP) in human retinal microvascular endothelial cells (HRMECs) was evaluated using qPCR and the Seahorse XFe96 analyzer under normal culture conditions and hypoxia ± 3PO. The role of glycolysis in HRMEC angiogenesis related processes such as tubulogenesis, proliferation and migration were assessed when glycolysis was blocked by 3PO. Our study showed that intravitreal injection of 3PO in the OIR model inhibited pre-retinal neovascularization compared to vehicle injected controls (P < 0.0001). While hypoxia increased glycolysis in HRMECs, treatment with 3PO reduced their glycolytic activity under normoxia and hypoxia culture conditions. Treatment with 3PO, reduced glycolytic mRNA expression of GLUT1, HK1, PFKFB3, ENO2 and VEGFA. Finally, glycolytic inhibition reduced tubulogenesis (p < 0.05), migratory capacity (p < 0.001) and proliferation (p < 0.01) of HRMECs in vitro. This data suggests that retinal angiogenesis can be modulated by manipulating the glycolytic pathway using 3PO in vivo and that 3PO treatment in vitro can diminish the angiogenic potential of HRMECs.
在许多视网膜病变中，缺氧会刺激致病性新生血管形成。缺氧的确切影响以及在活跃血管生成过程中它如何驱动视网膜微血管内皮细胞代谢转变仍不明确。本研究旨在了解暴露于缺氧环境以及在缺血诱导的新生血管形成过程中视网膜微血管内皮细胞代谢谱的动态变化。使用氧诱导视网膜病变（OIR）模型，研究了通过玻璃体内注射糖酵解抑制剂3-(3-吡啶基)-1-(4-吡啶基)-2-丙烯-1-酮（3PO）来操纵糖酵解代谢对视网膜前新生血管形成的影响。在正常培养条件以及缺氧±3PO条件下，使用qPCR和海马XFe96分析仪评估人视网膜微血管内皮细胞（HRMECs）中产生三磷酸腺苷（ATP）的代谢途径。当糖酵解被3PO阻断时，评估糖酵解在HRMEC血管生成相关过程（如形成管状结构、增殖和迁移）中的作用。我们的研究表明，在OIR模型中，与注射赋形剂的对照组相比，玻璃体内注射3PO可抑制视网膜前新生血管形成（P < 0.0001）。虽然缺氧会增加HRMECs中的糖酵解，但在常氧和缺氧培养条件下，用3PO处理会降低它们的糖酵解活性。用3PO处理可降低GLUT1、HK1、PFKFB3、ENO2和VEGFA的糖酵解mRNA表达。最后，糖酵解抑制降低了体外HRMECs的形成管状结构能力（p < 0.05）、迁移能力（p < 0.001）和增殖能力（p < 0.01）。这些数据表明，在体内使用3PO操纵糖酵解途径可调节视网膜血管生成，并且体外3PO处理可降低HRMECs的血管生成潜力。
