ELF3-regulated PES1 targets VEGFR2 to mediate angiogenesis and retinal inner barrier injury in diabetic retinopathy

Journal of Translational Medicine Liu et al. Journal of Translational Medicine (2025) 23:1309 https://doi.org/10.1186/s12967-025-07334-0 Open Access RESEARCH ELF3-regulated PES1 targets VEGFR2 to mediate angiogenesis and retinal inner barrier injury in diabetic retinopathy Sha Liu1,2†, Jingjing Hou1,2†, Yulin Tao1, Yi Xu1,2, Jinghui Liu1, Siyu Lin1,2, Liming Tao1* and Zhengxuan Jiang1* Abstract Background Angiogenesis and increased vascular permeability caused by endothelial cell dysfunction are crucial in diabetic retinopathy (DR). Pescadillo ribosomal biogenesis factor 1 (PES1) plays key roles in diabetes; however, its effect on DR has not been thoroughly investigated. This study evaluated the role of PES1 in angiogenesis and bloodretinal barrier functions of DR and explored the related regulatory mechanisms. Methods Integrated bulk RNA-seq and single-cell RNA-seq analyses were used to identify the key molecular target PES1 in this study. Western blot, scratch assay, tube formation assay, transwell, EdU proliferation assay, immunofluorescence, Evans Blue assay, PAS staining and H&E staining were used to detect the role of PES1 in angiogenesis and retinal barrier injury in vivo and in vitro, respectively. ChIP-seq was used to explore the regulatory mechanism. Results Integrated bulk RNA-seq and single-cell RNA-seq analyses revealed that high PES1 expression was significantly associated with DR. Under hyperglycemia conditions, elevated PES1 expression disrupted the retinal vascular barrier function and exacerbated vascular leakage and DR progression by inhibiting the expression of VE-cadherin and Occludin. Moreover, PES1 aggravated the imbalance between oxidation and anti-oxidation, leading to an excessive release of reactive oxygen species and further impairment of endothelial cell function. These biological processes were reversed by PES1-targeted siRNA/shRNA. Mechanistically, PES1 bound to the enhancer of VEGFR2 to regulate VEGFR2 mRNA expression, thereby influencing tube formation capacity and permeability of endothelial cells. Under diabetic conditions, PES1 upregulation was regulated by ELF3 binding to its promoter. Conclusions Our study confirmed that the ELF3/PES1/VEGFR2 signaling pathway is crucial for regulating retinal angiogenesis and blood-retinal barrier function in DR. Interventions targeting ELF3 or PES1 may serve as potential therapeutic strategies for DR. Keywords PES1, VEGFR2, ELF3, Endothelial cell dysfunction, Diabetic retinopathy † Sha Liu and Jingjing Hou contributed equally to this work. *Correspondence: Liming Tao Zhengxuan Jiang 1 Department of Ophthalmology, The Second Affiliated Hospital of Anhui Medical University, Anhui Medical University, Hefei, Anhui Province 230000, China 2 Department of Clinical Medicine, The Second School of Clinical Medicine, Anhui Medical University, Hefei, Anhui Province 230022, China © The Author(s) 2025. Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creati vecommons.org/licenses/by-nc-nd/4.0/. Liu et al. Journal of Translational Medicine (2025) 23:1309 Introduction Diabetic retinopathy (DR) is the main microvascular complication of diabetes with a notably high incidence rate. With the increasing number of patients with diabetes, DR has become a major, irreversible blinding eye disease among working-age and older individuals [1]. Pathological angiogenesis caused by vascular endothelial injury is the initial event in DR [2]. Although anti-VEGF signaling pathway drugs are first-line therapies for DR, they exhibit unsatisfactory efficacy in some patients, leading to therapeutic resistance [3]. Hence, there is an urgent need to elucidate the mechanisms underlying the progression of DR and identify novel anti-VEGF signaling pathway regulators, to control pathological angiogenesis and develop novel strategies for DR treatment. Pescadillo ribosomal biogenesis factor 1 (PES1), a nucleolar protein with nuclear localization signals, acts as a transcription factor (TF) to promote the transcription of genes required for ribosome biogenesis [4, 5]. PES1 interacts with various proteins, affects the cell cycle by activating various pathways, and plays important roles in cell proliferation, senescence, and autophagy [6–8]. Additionally, PES1 is an important factor in stabilizing ribosome biogenesis [9]. The biogenesis of ribosomes is necessary for angiogenesis [10], implying that PES1 may be involved in angiogenesis. Notably, it has been shown that the ablation of PES1 can inhibit VEGF signaling pathway-mediated angiogenesis in gastric cancer [11]. Moreover, increasing evidence indicates that PES1 plays a role in the pathogenesis of metabolic diseases. Overexpression of PES1 promotes lipid dysregulation and glycolysis in diabetes [12, 13], and its role in regulating vascular permeability in diabetes is well established [14]. Therefore, PES1 is potentially correlated with DR development; however, the functions and regulatory mechanisms of PES1 in DR remain unexplored. In the present study, a critical role of PES1 in the vascular endothelial dysfunction of DR was revealed. Our results demonstrated that the VEGF/VEGFR2 signaling pathway was significantly activated by PES1, manifesting as a disruption of the blood-retinal barrier and neovascularization. The facilitatory role of PES1 in VEGFR2 signaling is mediated through interactions with the VEGFR2 enhancer. Notably, we found that PES1 was regulated by ELF3, thus playing a crucial role in DR. This was accomplished by recruiting ELF3 to the promoter of PES1, thereby promoting PES1 transcription. Increased PES1 expression in DR may serve as a potential biomarker for the aggravation of microvascular complications. Materials and methods Single-cell and RNA sequencing analysis The datasets GSE102485, GSE160306, GSE165784, GSE245561, GSE135922, and GSE178121 from the GEO Page 2 of 17 database were downloaded (https://www.ncbi.nlm.nih.g ov/geo/) [15–20]. DR patients with proliferative diabetic retinopathy (PDR) or diabetic macular edema (DME) and control (...truncated)