Aug 2026· Theranostics· Vol 16, pp. 8650-8668· 0 citations· 84 references
TL;DR
In vivo rescue experiments confirmed that ETS1 overexpression reversed the suppression of pathological neovascularization in OIR mice treated with AAV-Pfkfb3-RNAi, providing a promising therapeutic avenue for ischemic retinal diseases.
Abstract
Ocular neovascular disease, characterized by aberrant angiogenesis in the eye, is a primary cause of global vision impairment and blindness. As the primary barrier exposed to hypoxia-related blood metabolites, endothelial cells (ECs) undergo metabolic reprogramming that drives pathological angiogenesis. However, the epigenetic mechanisms that link EC metabolic dysfunction to retinal vasculopathy remain elusive. Methods: Using western blotting and immunofluorescence analysis of retinal sections/whole-mounts, we confirmed increased histone 3 lactylation at lysine 18 (H3K18la). We subsequently identified downstream target genes through integrated CUT&Tag and RNA sequencing (RNA-seq), assessed their angiogenic regulatory functions using siRNA, and validated the mechanisms in vivo employing adeno-associated virus (AAV)-based gene transfer. Results: Our data indicated that histone lactylation levels were elevated in retinal vascular ECs under hypoxic conditions both in vivo and in vitro . In oxygen-induced retinopathy (OIR) retinal vascular ECs, H3K18la was the most prominent modification. Pharmacological inhibition of glycolysis suppressed H3K18la levels, concurrently abrogating EC activation and neovascularization. Combined CUT&Tag and RNA-seq analyses revealed that ETS1 was a direct transcriptional target governed by H3K18la in retinal ECs. Silencing ETS1 substantially inhibited hypoxia-induced proliferation, migration, sprouting, and tube formation in human retinal microvascular endothelial cells (HRMECs). Crucially, in vivo rescue experiments confirmed that ETS1 overexpression reversed the suppression of pathological neovascularization in OIR mice treated with AAV-Pfkfb3-RNAi. Conclusions: Collectively, this study revealed a lactate-driven epigenetic cascade wherein H3K18la licenses ETS1-dependent pathological angiogenesis, providing a promising therapeutic avenue for ischemic retinal diseases.
The cKIAA1462/miR-183-5p/HMGB1 axis plays a critical role in diabetic RPE injury by dual regulation of autophagy and pyroptosis by targeting this pathway may offer a novel therapeutic strategy for early diabetic retinopathy.
Lufei Wang, Jia’nan Xie, Longfei Yang et al.· Journal of Translational Med...· 0 citations
Mechanistically, circPSEN1 acts as a sponge for miR-150-5p, enhancing TRIM65 expression and contributing to DR pathogenesis, indicating that circPSEN1 may serve as a therapeutic target for DR.
Pathological ocular neovascularization remains a leading cause of irreversible vision loss. This study investigates the role of Polo-like kinase 2 (PLK2) in coordinating the transition of endothelial cells into pathological phenotypes. Single-cell transcriptomic and clinical tissue analysis revealed that PLK2 is specifically enriched in the vascular component of human proliferative membranes and is markedly upregulated in human fibrovascular membrane tissue. PLK2 endothelial knockdown (eKD) inhibited vascular branching and tip cell filopodia formation in postnatal mice. Analysis of murine oxygen-induced retinopathy (OIR) single cell sequencing datasets identified PLK2 as a specific marker for actively sprouting tip cells and proliferative endothelial subpopulations during pathological neovascularization. PLK2 eKD or conditional knockout (eCKO) significantly attenuated pathological neovascularization in OIR. Additionally, PLK2 eKD inhibited laser-induced choroidal neovascularization (CNV) in adult mice. In vitro assays confirmed that PLK2 depletion via shRNA or CRISPR/Cas9 markedly inhibited endothelial cell proliferation, migration, and tube formation across various microvascular cells, whereas ectopic PLK2 overexpression promoted a robust pro-angiogenic phenotype. Mechanistically, PLK2 is a critical regulator of mitochondrial bioenergetics; genetic depletion led to a reduction of mitochondrial Complex I activity, decreased ATP levels, and impaired oxygen consumption, accompanied by mitochondrial membrane depolarization and the accumulation of reactive oxygen species (ROS). The anti-angiogenic effects and apoptotic induction caused by PLK2 deficiency were significantly rescued by glucose supplementation or antioxidant N-acetylcysteine (NAC) treatment. Thus, by maintaining mitochondrial bioenergetic efficiency and mitigating oxidative stress, PLK2 sustains the high-energy demands of pathological endothelial cell activation and sprouting. PLK2 represents a promising therapeutic strategy for pathological ocular angiogenesis.
Ping-ping Fu, Wen Bai, Ying-shun Pan et al.· Cell Death Discovery· 0 citations
Current research on pathological retinal neovascularization primarily focuses on growth factors, inflammation, and endothelial signaling pathways. However, increasing attention is being directed toward disruptions in the retinal immune microenvironment. Therefore, deciphering the immune-angiogenic interplay could uncover therapeutic avenues for neovascular disorders. Through integrative analyses of single-cell RNA sequencing from human fibrovascular membranes and multicohort clinical datasets, we identified neutrophil infiltration as an independent risk factor for diabetic retinopathy progression. Mechanistically, activated microglia preceded and potentiated neutrophil infiltration by secreting galectin-3 (GAL3), establishing a self-amplifying feedback loop that sustained microglial activation and drove pathological angiogenesis in mice with oxygen-induced retinopathy (OIR). To therapeutically disrupt this loop, we engineered a photocurable hydrogel for the sustained intravitreal delivery of GAL3 and vascular endothelial growth factor (VEGF)-neutralizing antibodies, which effectively suppressed aberrant angiogenesis in mice with OIR. Together, these findings reinforce the concept of retinal neovascularization as an immunovascular disorder and underscore the therapeutic potential of microenvironment-modulating strategies using biomaterials.
Ziyi Zhou, Tianhao Yuan, Jiaxing Sun et al.· Science Translational Medici...· 0 citations
Background Pulmonary hypertension (PH) involves progressive vascular remodeling and perivascular inflammation. Despite modest clinical improvements with current therapies, their limited ability to reverse remodeling or restore immune homeostasis highlights the need for novel agents. Liriodendrin (Lidd), derived from Sargentodoxae caulis, exhibits anti-inflammatory and antiproliferative activities, but its efficacy and molecular targets in PH are unknown. Methods Two well-established PH animal models - the SU5416/hypoxia (SuHx) mice model and monocrotaline (MCT)-induced rat model - were employed for in vivo assessment of Lidd conducted pharmacological effects. Primary human pulmonary artery smooth muscle cells (hPASMCs) were utilized for mechanistic investigations. RNA-sequencing (RNA-seq) analysis was conducted to identify potential signaling pathways modulated by Lidd treatment. The direct molecular target of Lidd was determined through integrated application of drug affinity responsive target stability (DARTS) assay coupled with western blot validation. To delineate histone lactylation-mediated transcriptional regulation, we combined Cleavage Under Targets and Tagmentation (CUT&Tag) sequencing data analysis followed by chromatin immunoprecipitation quantitative PCR (ChIP-qPCR) verification. Genetic validation was achieved using PFKFB3-deficient murine models to verify the specificity of Lidd-mediated pharmacological actions. Results Lidd administration attenuated pulmonary vascular remodeling, perivascular macrophage infiltration and PH progression in both SuHx and MCT models. Transcriptomic profiling of Lidd-treated hPASMCs revealed predominant enrichment of downregulated genes in inflammatory and cytokine-associated pathways. Mechanistically, Lidd directly bound PFKFB3 and enhanced its interaction with FZR1, promoting PFKFB3 ubiquitination and degradation, which reduced glycolysis-driven lactate and consequent histone lactylation. This, in turn, diminished transcriptional activation of proliferative and inflammatory mediators, including CCND1, TNC, and CCL2. Notably, exogenous lactate supplementation or endogenous lactate accumulation restored histone lactylation and paradoxically potentiated Lidd’s inhibitory effects on PASMC proliferation and migration, whereas p300 inhibition abrogated these lactate-mediated effects. Importantly, Lidd failed to confer additional protection in PFKFB3-deficient mice, confirming PFKFB3 as the primary target mediating its therapeutic action. Conclusion Our findings reveal that Lidd selectively targets the PFKFB3-mediated glycolytic-epigenetic axis to suppress PASMC phenotypic transformation and pulmonary vascular remodeling, positioning it as a promising therapeutic candidate for PH.
Qingye Zeng, Zhenzhen Duan, Qian Liu et al.· bioRxiv· 0 citations
Background Diabetic retinopathy (DR) is a leading cause of preventable vision loss worldwide, characterized by retinal microvascular dysfunction and progressive angiogenic dysregulation. Angiopoietin-like protein 2 (ANGPTL2) promotes endothelial activation and pathological angiogenesis in multiple vascular disease contexts, yet its effects on retinal endothelial angiogenic behavior have not been directly characterized. Whether integrin α5β1 and PI3K/AKT phosphorylation changes in the diabetic retina are associated with alterations in ANGPTL2 expression levels also remains unexamined. Methods ANGPTL2 was overexpressed in human retinal microvascular endothelial cells (HRMECs), and effects on cell viability, wound closure, invasion, and tube formation were assessed by CCK-8, wound healing, Transwell invasion, and tube formation assays. Integrin α5β1, VEGF, and PI3K/AKT phosphorylation were characterized by western blot. Retinal tissues from streptozotocin (STZ)-induced diabetic rats across groups with varying ANGPTL2 expression levels were analyzed by RT-qPCR, western blot, and immunofluorescence to characterize accompanying molecular changes. The protein-level association between ANGPTL2 and integrin α5β1 was examined by co-immunoprecipitation and co-localization immunofluorescence under overexpression conditions. Results ANGPTL2 overexpression significantly enhanced cell viability, wound closure, invasion, and tube formation in HRMECs, accompanied by upregulation of VEGF, p-VEGFR2, integrin α5β1, and PI3K/AKT phosphorylation. Retinal tissues from STZ-diabetic rats showed elevated integrin α5β1 expression, increased CD31 levels, and increased PI3K/AKT phosphorylation relative to non-diabetic controls; these molecular changes were directionally lower in the ANGPTL2 knockdown comparison group (i.e., the STZ + sh-ANGPTL2 group), providing supportive retinal molecular context. Co-immunoprecipitation and co-localization analyses indicated a protein-level association between ANGPTL2 and integrin α5β1 under overexpression conditions. Conclusion ANGPTL2 overexpression promotes angiogenic responses in retinal endothelial cells in vitro, with accompanying integrin α5β1 upregulation and broader signaling-associated phosphorylation changes. Retinal tissue molecular findings and the ANGPTL2–integrin α5β1 protein-level association provide supportive molecular context. The functional roles of specific molecular partners in mediating these responses remain to be determined through targeted validation studies.
Jiajia Song, Xiaofang Han, Jianfeng Chen et al.· Frontiers in Medicine· 0 citations