Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore 636921, Singapore.
Duke-NUS Medical School, National University of Singapore, Singapore 169857, Singapore.
Int J Mol Sci. 2022 Oct 19;23(20):12549. doi: 10.3390/ijms232012549.
Age-related macular degeneration (AMD) is a global leading cause of visual impairment in older populations. 'Wet' AMD, the most common subtype of this disease, occurs when pathological angiogenesis infiltrates the subretinal space (choroidal neovascularization), causing hemorrhage and retinal damage. Gold standard anti-vascular endothelial growth factor (VEGF) treatment is an effective therapy, but the long-term prevention of visual decline has not been as successful. This warrants the need to elucidate potential VEGF-independent pathways. We generated blood out-growth endothelial cells (BOECs) from wet AMD and normal control subjects, then induced angiogenic sprouting of BOECs using a fibrin gel bead assay. To deconvolute endothelial heterogeneity, we performed single-cell transcriptomic analysis on the sprouting BOECs, revealing a spectrum of cell states. Our wet AMD BOECs share common pathways with choroidal neovascularization such as extracellular matrix remodeling that promoted proangiogenic phenotype, and our 'activated' BOEC subpopulation demonstrated proinflammatory hallmarks, resembling the tip-like cells in vivo. We uncovered new molecular insights that pathological angiogenesis in wet AMD BOECs could also be driven by interleukin signaling and amino acid metabolism. A web-based visualization of the sprouting BOEC single-cell transcriptome has been created to facilitate further discovery research.
年龄相关性黄斑变性（AMD）是导致老年人群视力损害的全球主要原因。湿性 AMD 是这种疾病最常见的亚型，当病理性血管生成渗透到视网膜下空间（脉络膜新生血管形成）时发生，导致出血和视网膜损伤。抗血管内皮生长因子（VEGF）的金标准治疗是一种有效的治疗方法，但长期预防视力下降的效果并不理想。这就需要阐明潜在的 VEGF 非依赖性途径。我们从湿性 AMD 和正常对照组中生成了血液外生内皮细胞（BOECs），然后使用纤维蛋白凝胶珠测定法诱导 BOECs 的血管生成发芽。为了解内皮细胞异质性，我们对发芽的 BOECs 进行了单细胞转录组分析，揭示了一系列细胞状态。我们的湿性 AMD BOECs 与脉络膜新生血管形成共享共同的途径，如促进促血管生成表型的细胞外基质重塑，而我们的“激活”BOEC 亚群表现出促炎特征，类似于体内的尖端样细胞。我们揭示了新的分子见解，即湿性 AMD BOECs 的病理性血管生成也可能受到白细胞介素信号和氨基酸代谢的驱动。已经创建了一个基于网络的发芽 BOEC 单细胞转录组可视化工具，以促进进一步的发现研究。
