Ding Xiaoyan, Zhou Xiaodi, Liu Xinyu, Lai Yanting, Yan Wenjia, Cheng Yizhe, Hou Aohan, Chen Limei, Sun Limei
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, China.
Invest Ophthalmol Vis Sci. 2025 Feb 3;66(2):40. doi: 10.1167/iovs.66.2.40.
Neovascular eye diseases, such as proliferative diabetic retinopathy (PDR), wet age-related macular degeneration (wAMD) and retinopathy of prematurity (ROP), are major causes of vision loss and blindness worldwide. Our transcription factor motif enrichment analysis highlighted RUNX1 as a key regulator in the hypoxic response. The purpose of this study was to characterize how loss of Runx1 affects physiological and pathological retinal vasculature formation.
RNA-seq analysis and Transcription factor motif enrichment analysis were conducted in hypoxic and normoxic HUVECs. Conditional deletion of Runx1 in endothelial cells In mice was achieved using recombinase driver Cdh5-CreERT2. Vascular coverage, density, vessel progression, branchpoints, and sprout numbers was measured in retina of Runx1iECKO mice. The expression patterns, functions, and potential therapeutic value of RUNX1 were further explored with clinical samples, as well as in vivo and in vitro experiments. Bioinformatics and high-throughput sequencing were performed to identify potential target genes of Runx1. RT-qPCR and Western blot analyses were carried out to detect the changes of PI3-kinase/AKT/mTOR pathway.
Loss of Runx1 in mice resulted in a reduction of the vascular coverage, density, vessel progression, branchpoints, and sprouts numbers of the retinal vascular network during its development. Notably, mature blood vessels remained unaffected by Runx1 inhibition. Upregulation of RUNX1 was observed in patients with PDR and ROP. RUNX1 Inhibition reduced endothelial cell proliferation, migration and tubule formation, leading to decreased pathological neovascularization, which is shown in oxygen-induced retinopathy. Mechanistically, in vitro experiments demonstrated that RUNX1 regulates EC angiogenesis through the PI3K/AKT/mTOR signaling pathway.
Runx1 is essential for physiological retinal vascularization. RUNX1 Inhibition may effectively decrease pathological neovascularization. Our findings suggest that targeting RUNX1 could be a promising therapeutic strategy for retinal neovascular disorders, preserving the integrity of mature blood vessels while selectively inhibiting neovascularization.
新生血管性眼病，如增殖性糖尿病视网膜病变（PDR）、湿性年龄相关性黄斑变性（wAMD）和早产儿视网膜病变（ROP），是全球视力丧失和失明的主要原因。我们的转录因子基序富集分析突出显示RUNX1是缺氧反应中的关键调节因子。本研究的目的是表征Runx1缺失如何影响生理性和病理性视网膜血管形成。
在缺氧和常氧的人脐静脉内皮细胞（HUVECs）中进行RNA测序分析和转录因子基序富集分析。使用重组酶驱动因子Cdh5-CreERT2在小鼠内皮细胞中实现Runx1的条件性缺失。在Runx1iECKO小鼠的视网膜中测量血管覆盖率、密度、血管进展、分支点和新芽数量。通过临床样本以及体内和体外实验进一步探索RUNX1的表达模式、功能和潜在治疗价值。进行生物信息学和高通量测序以鉴定Runx1的潜在靶基因。进行逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹分析以检测PI3激酶/AKT/雷帕霉素靶蛋白（mTOR）通路的变化。
小鼠中Runx1的缺失导致视网膜血管网络在发育过程中血管覆盖率、密度、血管进展、分支点和新芽数量减少。值得注意的是，成熟血管不受Runx1抑制的影响。在PDR和ROP患者中观察到RUNX1上调。RUNX1抑制减少内皮细胞增殖、迁移和小管形成，导致病理性新生血管形成减少，这在氧诱导的视网膜病变中得到体现。从机制上讲，体外实验表明RUNX1通过PI3K/AKT/mTOR信号通路调节内皮细胞血管生成。
Runx1对生理性视网膜血管化至关重要。RUNX1抑制可能有效减少病理性新生血管形成。我们的研究结果表明，靶向RUNX1可能是治疗视网膜新生血管疾病的一种有前景的治疗策略，在选择性抑制新生血管形成的同时保持成熟血管的完整性。
