Targeting long non-coding RNA RP11-502I4.3 delays the trend of angiogenesis in diabetic retinopathy
Lan Zeng, Yuhao WuiD, Lijuan ZhuiD, Junhao He, Yuan YuaniD, Xiaocong WangiD, Kai TangiD, Wei TaniD
Abstract
Based on our previous findings, we hypothesized that the long non-coding RNA RP11-502I4.3 may be involved in angiogenesis associated with Diabetic retinopathy (DR). We investigated the role of RP11-502I4.3 in DR by examining its regulation of vascular endothelial growth factor (VEGF). We assessed differences in RP11-502I4.3 expression between the normal control group and streptozotocin-induced diabetic rats or high glucose (HG)-stimulated human retinal microvascular endothelial cells (HRMECs). VEGF expression was measured with and without lentiviral vectors overexpressing RP11-502I4.3. We analyzed the structural and functional alterations related to DR. Our analysis revealed that RP11-502I4.3 expression was lower in the retinas of diabetic rats and in HG-stimulated HRMECs compared with normal glucose conditions. Overexpressing of RP11-502I4.3 resulted in decreased VEGF levels. Diabetic rats exhibited retinopathy characterized by thinning of the retinal layer thickness, structural changes in the inner and outer nuclear layers, a reduced count of retinal ganglion cells, and the presence of acellular capillaries. The proliferative activity, migration count, and tube formation ability of HG-treated HRMECs were significantly higher than those of the normal control group; however, these changes were delayed by RP11-502I4.3 overexpression. RP11-502I4.3 downregulation in DR appears to promote angiogenesis.
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In-depth summary
Claude Sonnet 4.6 · xhigh effort · 8 Nov 2026 · from the full text of version 1 (18 Oct 2024) · CC BY 4.0
Lowered RP11-502I4.3 lncRNA expression in diabetic rat retinas and high-glucose endothelial cells; lentiviral overexpression reduced VEGF and attenuated angiogenesis-related phenotypes and retinopathy features.
Treatment outcomes in proliferative diabetic retinopathy are often suboptimal, and lncRNA function in this disease has received little attention. Starting from a microarray finding (GSE191210) of lower RP11-502I4.3 in patient vitreous, the authors test whether it restrains VEGF-linked angiogenesis.
Key results
ShownRP11-502I4.3 is downregulated across three diabetic-retinopathy contexts: patient vitreous humor, retinas of streptozotocin-induced diabetic rats, and high-glucose-treated human retinal microvascular endothelial cells (HRMECs). VEGF mRNA is increased in diabetic rat retina.
Vitreous: PDR vs idiopathic macular hole, microarray GSE191210 confirmed by qRT-PCR. Rats: 12 weeks after induction, eight diabetic vs eight control rats. HRMECs: 30 mmol/L glucose for 48 hours. Only 'significantly lower' is reported; no effect sizes. · Differences in RP11-502I4.3 expression in a…; Abstract; Cells culture; Animals
ShownLentiviral overexpression of RP11-502I4.3 lowered VEGF expression in the retinas of diabetic rats and in high-glucose HRMECs, relative to overexpression-control vectors.
Rats: right eyes (overexpression, 16 rats) vs left eyes (control vector, 16 rats). HRMEC transfection efficiency 70-75% (exceeding 70% by GFP). VEGF 'significantly downregulated' in both models; no fold-changes or p-values given. · RP11-502I4.3 Overexpression delayed the trend of…; Establishment of RP11-502I4.3 overexpression…; Cells transfection; Animals
SuggestedDiabetic rats developed retinopathy features (thinner retinal layers, inner and outer nuclear layer changes, fewer retinal ganglion cells, acellular capillaries). Overexpression delayed these changes and reduced acellular capillary counts.
Diabetes confirmed by blood glucose above 16.7 mmol/L; tissues collected 12 weeks after induction; H&E and PAS staining. Diabetic vs normal groups were eight rats each, overexpression vs control 16 rats each. Rescue effects are described qualitatively, without numerical values. · Differences in RP11-502I4.3 expression in a…; RP11-502I4.3 Overexpression delayed the trend of…; Animals; Haematoxylin and eosin (H&E) staining
SuggestedHigh glucose increased HRMEC proliferation, migration and tube formation, and RP11-502I4.3 overexpression delayed all three changes.
CCK-8 (450 nm), Transwell migration counts and Matrigel tube formation assays; each repeated three times in parallel. Described as 'significantly higher' under high glucose and delayed by overexpression; no effect sizes reported. · Abstract; RP11-502I4.3 Overexpression delayed the trend of…; Cell Counting Kit-8 (CCK-8) assay; Transwell assay
ClaimedThe authors propose that RP11-502I4.3 loss promotes retinal neovascularisation by regulating VEGF. Evidence is limited to VEGF expression changing with overexpression; no mechanism is tested.
Conclusion rests on lower VEGF after overexpression in rats and HRMECs. The text reports no knockdown, rescue, binding or pathway experiments, and the lncRNA is too poorly annotated to predict targets bioinformatically. · Discussion; Limitations; Conclusions
Methods, data and reproducibility
Expression of RP11-502I4.3 and VEGF was compared by qRT-PCR between normal and high-glucose/diabetic conditions, with and without lentiviral overexpression, in rat retina and HRMECs. Retinal histology and endothelial proliferation, migration and tube formation were read out.
- Vitreous humor lncRNA microarray, proliferative diabetic retinopathy vs idiopathic macular hole (prior analysis), confirmed by qRT-PCR · human · GSE191210
- Streptozotocin-induced diabetic Sprague-Dawley rats (high-fat diet plus streptozotocin) vs normal controls; both eyes used per rat · rat · n = eight rats each for normal and diabetes groups
- Diabetic rats with intravitreal lentiviral RP11-502I4.3 overexpression (right eye) or control vector (left eye) · rat · n = 16 rats per overexpression group
- HRMEC cultures: normal, high glucose, overexpression control, overexpression (in vitro assays) · human · n = experiments repeated three times in parallel
- Methods & tools
- qRT-PCR (2^-ΔΔCt, β-actin control), Lentiviral overexpression vector, H&E staining, PAS staining of isolated retinal vessels, Fluorescence in situ hybridization, CCK-8 assay, Transwell migration assay, Matrigel tube formation assay, ImageJ 1.8.0, GraphPad Prism 8, t-test, one-way ANOVA, rank-sum test
- Reagents & environment
- ImageJ 1.8.0 and GraphPad Prism 8 for analysis; no other software environment is described.
- Reproducibility gaps
- Datasets are available only from the corresponding author on reasonable request; no code or repository URLs are given.
Limitations (5)
- RP11-502I4 is poorly annotated, so associated mRNAs and signaling pathways could not be predicted bioinformatically; the regulatory mechanism remains unresolved. · stated by the authors
- Only endothelial cells (HRMECs) were used in vitro, so effects on pericytes and other retinal cells, and their interactions, were not examined. · stated by the authors
- Rats do not fully reproduce DR features such as microaneurysms and neovascularisation, and clinical applicability of lncRNA therapy is untested. · stated by the authors
- Only overexpression was tested, with no knockdown or VEGF rescue experiments, so the link between the lncRNA and VEGF is correlative. · noted in this summary
- The text reports results mainly as significant differences, without numerical effect sizes or specific p-values, which limits quantitative appraisal of the findings. · noted in this summary
Who should read:Read this if you study lncRNA control of VEGF-linked angiogenesis in diabetic retinopathy and want rat and endothelial-cell overexpression evidence.
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