Biswas Nilima, Mori Tommaso, Ragava Chetty Nagaraj Naresh Kumar, Xin Hong, Diemer Tanja, Li Pin, Su Yongxuan, Piermarocchi Carlo, Ferrara Napoleone
Department of Pathology, University of California San Diego, La Jolla, CA 92093.
Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA 92093.
Proc Natl Acad Sci U S A. 2025 Jan 28;122(4):e2418752122. doi: 10.1073/pnas.2418752122. Epub 2025 Jan 21.
We hypothesized that a strategy employing tissue-specific endothelial cells (EC) might facilitate the identification of tissue- or organ-specific vascular functions of ubiquitous metabolites. An unbiased approach was employed to identify water-soluble small molecules with mitogenic activity on choroidal EC. We identified adenosine diphosphate (ADP) as a candidate, following biochemical purification from mouse EL4 lymphoma extracts. ADP stimulated the growth of bovine choroidal EC (BCEC) and other bovine or human eye-derived EC. ADP induced rapid phosphorylation of extracellular signal-regulated kinase in a dose- and time-dependent manner. ADP-induced BCEC proliferation could be blocked by pretreatment with specific antagonists of the purinergic receptor P2Y1 but not with a vascular endothelial growth factor (VEGF) inhibitor, indicating that the EC mitogenic effects of ADP are not mediated by stimulation of the VEGF pathway. Intravitreal administration of ADP expanded the neovascular area in a mouse model of choroidal neovascularization. Single-cell transcriptomics from human choroidal datasets show the expression of P2RY1, but not other ADP receptors, in EC with a pattern similar to VEGFR2. Although ADP has been reported to be a growth inhibitor for vascular EC, here we describe its growth-stimulating effects for BCEC and other eye-derived EC.
我们假设，采用组织特异性内皮细胞（EC）的策略可能有助于识别普遍存在的代谢物的组织或器官特异性血管功能。我们采用一种无偏倚的方法来鉴定对脉络膜EC具有促有丝分裂活性的水溶性小分子。从小鼠EL4淋巴瘤提取物中进行生化纯化后，我们鉴定出二磷酸腺苷（ADP）为候选物。ADP刺激牛脉络膜EC（BCEC）以及其他牛或人眼源性EC的生长。ADP以剂量和时间依赖性方式诱导细胞外信号调节激酶的快速磷酸化。用嘌呤能受体P2Y1的特异性拮抗剂预处理可阻断ADP诱导的BCEC增殖，但血管内皮生长因子（VEGF）抑制剂则不能，这表明ADP对EC的促有丝分裂作用不是通过刺激VEGF途径介导的。在脉络膜新生血管小鼠模型中，玻璃体内注射ADP可扩大新生血管区域。来自人类脉络膜数据集的单细胞转录组学显示，在EC中P2RY1而非其他ADP受体的表达模式与VEGFR2相似。尽管据报道ADP是血管EC的生长抑制剂，但在此我们描述了其对BCEC和其他眼源性EC的生长刺激作用。
