Dimethyl Fumarate (DMFu) as an anti-angiogenic agent in VEGF-treated human microvascular retinal endothelial cells (HRECs).
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE285562 | GEO | 2025/09/03
REPOSITORIES: GEO
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Project description: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 (https://christinecheunglab.shinyapps.io/human_wet_AMD_sprouting/) has been created to facilitate further discovery research.
2022-11-02 | GSE213933 | GEO
Project description:Dysfunction of the blood retinal barriers and/or proliferation of retinal and choroidal endothelial cells are caused by late stages of diabetic retinopathy (DR) and neovascular age-related macular degeneration (nAMD). To elucidate endothelial-derived pathomechanisms in DR and nAMD, we established immortalized mouse cell lines of retinal (REC), choroidal (ChEC) and brain (BEC)endothelial cells. We compared the functional and transcriptomic state of these cells depending on their tissue origin. Intriguingly, activation of the wingless-type MMTV integration site (Wnt)/β-catenin signaling pathway restored barrier properties in BEC and REC, but increased permeability of ChEC. Transcriptome profiling showed that the immune system and pathways such as hypoxia-inducible factor (HIF) 1/2α-, transforming growth factor (TGF) β- and vascular endothelial growth factor (VEGF) were differentially regulated among endothelial cells. These findings significantly increase the understanding of the vascular biology of endothelial cells, highlighting the fact that depending on their tissue origin, their contribution to vascular pathologies such as DR or nAMD may vary.
2024-12-05 | GSE254272 | GEO
Project description:Dimethyl Fumarate (DMFu) as an anti-angiogenic agent in VEGF-treated human microvascular retinal endothelial cells (HRECs).
| PRJNA1204363 | ENA
Project description:PURPOSE Diseases that involve choroidal or retinal endothelial vascular cells are leading causes of vision loss: age-related macular degeneration, retinal ischemic vasculopathies and non-infectious posterior uveitis. Proteins differentially expressed by these endothelial cell populations are potential drug targets. We used deep proteomics to define the molecular phenotype of human choroidal and retinal endothelial cells at the protein level. METHODS Choroidal and retinal endothelial cells were separately isolated from 5 human eye pairs by selection on CD31. Total protein was extracted and digested, and peptide fractions were analysed by reverse-phase liquid chromatography tandem mass spectrometry. Peptide sequences were assigned to fragment ion spectra and proteins were inferred using public protein databases. Protein abundance was determined by spectral counting. Protein expression in choroidal versus retinal endothelial cells was compared using edgeR package in R, and annotation enrichment analysis was performed. RESULTS Human choroidal or retinal endothelial cells expressed 5042 non-redundant proteins. Setting the false discovery rate at 5%, 498 proteins (14.4%) of 3454 quantifiable proteins with minimum mean spectral counts of 2.5 were differentially expressed between cell populations. Choroidal and retinal endothelial cells were enriched in angiogenic proteins, and retinal endothelial cells were also enriched in immunologic proteins. CONCLUSIONS This work demonstrates the protein heterogeneity of human choroidal and retinal vascular endothelial cells and provides multiple candidates for further study as novel treatments or drug targets for posterior eye diseases.
2018-03-26 | PXD005972 | Pride
Project description:Patients with neovascular AMD (nAMD) suffer vision loss from destructive angiogenesis, termed choroidal neovascularization (CNV). Macrophages are found in CNV lesions from nAMD patients. Additionally, Ccr2-/- mice, which lack classical monocyte-derived macrophages, show reduced CNV size. However, macrophages are highly diverse cells that can perform multiple functions. We performed single-cell RNA-sequencing on immune cells from wildtype and Ccr2-/- eyes to uncover macrophage heterogeneity during the laser-induced CNV mouse model of nAMD. We identified 12 macrophage subtypes, including Spp1+ macrophages. Spp1+ macrophages were enriched from wildtype lasered eyes and expressed a pro-angiogenic transcriptome via multiple pathways, including vascular endothelial growth factor signaling, endothelial cell sprouting, cytokine signaling, and fibrosis. Additionally, Spp1+ macrophages expressed the marker CD11c, and CD11c+ macrophages were increased by laser and present in CNV lesions. Finally, CD11c+ macrophage depletion reduced CNV size by 40%. These findings broaden our understanding of ocular macrophage heterogeneity and implicate CD11c+ macrophages as a potential therapeutic target for treatment-resistant nAMD patients.
2023-02-27 | GSE222094 | GEO
Project description:Primary objectives: It is the primary objective of this study to assess whether there is a change and a correlation between ocular blood flow parameters (retinal vessel diameters, choroidal blood flow, fundus pulsation amplitude, retinal blood velocity and thickness) and treatment response in mCRC treated with standard of care anti-angiogenic regimens in combination with chemotherapy.
Primary endpoints: Retinal vessel diameters using The Retinal Vessel AnalyzerChoroidal blood flow using Laser Doppler flowmetryRetinal blood velocity by laser Doppler velocimeterFundus pulsation amplitude using laser interferometryRetinal thickness by OCT Plasma level of sVEGFR, VEGF, Tie2, intraocular pressure
| 2532658 | ecrin-mdr-crc
Project description:While quality of life compromising afflictions such as diabetic retinopathy (DR) are driven by vascular endothelial growth factor (VEGF), there is a growing appreciation that the concentration of VEGF is not the only influencer of vascular dysfunction within the retina. Activin A (activin), a ligand of the transforming growth factor β superfamily (TGF-β), attenuated VEGF-induced VE-cadherin disorganization, pore formation and permeability of primary human retinal endothelial cells (HRECs). Efforts to further investigate the mechanism of this phenomenon revealed that activin reduced expression of Rab11, which was required for the activin effect. The activin effect was not observed in cells with suppressed expression of the endosomally-localized protein tyrosine phosphatase, (PTP1b), whereas PTPs present on the plasma membrane were dispensable. Activin attenuated VEGF-mediated phosphorylation of VEGF receptor 2 (VEGFR2) at time points 30 min and longer post stimulation. Together these data support the concept that activin suppressed VEGF-induced barrier relaxation by perturbing trafficking of activated VEGFR2. This activin-based suppression of responsiveness to VEGF was compromised in the endothelium of pathological blood vessels from patients who developed end-stage proliferative diabetic retinopathy (PDR). This defect rendered HRECs hyper-responsive to VEGF and was not observed in retinal vessels of diabetic mice, which do not develop the angiogenic forms of DR. These data provide novel insights regarding the pathogenesis of PDR in patients.
2025-06-03 | GSE291008 | GEO
Project description:Neovascular age-related macular degeneration (nAMD) remains a major cause of visual impairment and puts considerable burden on patients and health care systems. L-DOPA-treated Parkinson Disease (PD) patients have been shown to be partially protected from nAMD, but the mechanism remains unknown. Using murine models combining 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD and laser-induced nAMD, standard PD treatment of L-DOPA/DOPA-decarboxylase inhibitor, or specific dopamine receptor inhibitors, we here demonstrate that L-DOPA treatment-induced increase of dopamine receptor D2 (DRD2) signaling inhibits choroidal neovascularization independently of MPTP-associated nigrostriatal pathway lesion. Analyzing a retrospective cohort of more than two hundred thousand nAMD patients receiving anti-VEGF treatment from the French nationwide insurance database, we show that DRD2-agonist treated (PD) patients have a significantly delayed age of onset for nAMD (81.4 (±7.0) vs 79.4 (±8.1) years old, respectively, p<0.0001) and reduced need for anti-VEGF therapies (-0.6 injections per 100 mg/day daily dose of DRD2 agonists the second year of treatment), similar to the L-DOPA treatment. While providing a mechanistic explanation for an intriguing epidemiological observation, our findings suggest that systemic DRD2 agonists might constitute an adjuvant therapy to delay and reduce the need for anti-VEGF therapy in nAMD patients.
2024-09-13 | GSE266525 | GEO
Project description:Background: Age-related macular degeneration (AMD) is the leading cause of blindness worldwide, predominantly affecting individuals over 60. AMD is classified into two clinical forms: dry (DAMD) and neovascular (NAMD). NAMD, characterized by choroidal neovascularization and increased vascular endothelial growth factor (VEGF) levels, is treated with anti-VEGF therapy, the gold standard for preventing vision loss. However, challenges including non-response to treatment, persistent disease activity, and unnecessary anti-VEGF administration necessitate the identification of predictive biomarkers to optimize therapy. Exosomes (exo) are extracellular vesicles known for their stability and role in cellular communication, carrying diverse molecular contents, including RNA. Although the role of exo-RNA in AMD pathogenesis is well known, their role in the response to anti-VEGF treatment has not yet been investigated. Current study aim to investigate the role of exo-RNAs as potential biomarkers for predicting anti-VEGF treatment response in NAMD patients. Methods: Plasma-derived exo-RNA were analyzed pre- and post-treatment to identify predictive biomarkers from 41 NAMD patients treated with intravitreal bevacizumab. Exo were isolated using exoRNAeasy midi kit and characterized by TEM, NTA, and Western blot. Then, RNA-seq was performed to profile exo-RNAs and key candidates were validated using RT-qPCR. Results: Following anti-VEGF therapy, 68.3% of NAMD patients (mean age: 70.8 ± 6.14), demonstrated anatomical success, while 53.7% achieved functional success. Based on RNA-seq comparative analysis between the two groups revealed that translation mechanisms were significantly regulated, particularly within the context of upregulated pathways. Genes associated with molecular functions such as insulin response, transport, angiogenesis, and oxygen transport were prominently implicated. Conversely, the evaluation of downregulated pathways indicated substantial alterations in RNA catabolic processes and the regulation of gene expression. RNA-seq revealed BMP6 and STK39 as key candidate genes, with post-treatment dysregulation in hsa-miR-4692 expression, a microRNA targeting STK39. Finally, RT-qPCR validation show that elevated pre-treatment exo-STK39 levels were significantly associated with anatomical success (*p=0.04).
2025-05-13 | GSE290090 | GEO
Project description:Consistent with clinical observations that posterior uveitis frequently involves the retinal vasculature and recent recognition of vascular heterogeneity, we hypothesized that retinal vascular endothelium was a cell population of unique molecular phenotype. Donor-matched cultures of primary retinal and choroidal endothelial cells from 6 human cadavers were incubated with either Toxoplasma gondii tachyzoites (10:1, parasite:cell) or Escherichia coli lipopolysaccharide (100 ng/mL); control cultures were simultaneously incubated with medium. Gene expression profiling of endothelial cells was performed using oligonucleotide arrays containing probes designed to detect 8747 human transcripts. After normalization, differential gene expression was assessed by the significance analysis of microarrays, with the false discovery rate set at 5 %. For selected genes, differences in level of expression between retinal and choroidal cells were evaluated by real-time RT-PCR. Graphical descriptive analysis demonstrated strong correlation between gene expression of unstimulated retinal and choroidal endothelial cells, but also highlighted distinctly different patterns of expression that were greater than differences noted between donors or between unstimulated and stimulated cells. Overall, 779 of 8,747 transcripts (8.9 %) were differentially represented. Notably, the 330 transcripts that were present at higher levels in retinal cells included a larger percentage of transcripts encoding molecules involved in the immune response. Differential gene expression was confirmed for 12 transcripts by RT-PCR. Retinal and choroidal vascular endothelial cells display distinctive gene expression profiles. Our findings suggest the possibility of treating posterior uveitis by targeting specific interactions between the retinal endothelial cell and an infiltrating leukocyte. Experiment Overall Design: Retinal and choroidal vascular endothelial cells were isolated from both eyes of 6 human cadaver donors. Endothelial cells of each subtype from each donor were separately pooled and cultured. In a first series of experiments (Experiment 1), donor-matched cultures of retinal and choroidal endothelial cells derived from 3 donors were incubated with either T. gondii tachyzoites or medium alone. In a second set of experiments (Experiment 2), cultured retinal and choroidal endothelial cells derived from the other 3 donors were stimulated with LPS or medium alone. Whenever possible, 2 replicate dishes of endothelial cells were subjected to each set of conditions. Subsequent to incubations, mRNA was extracted from a cell lysate prepared from each dish of cells, and cRNA derived from each mRNA preparation was separately hybridized to one oligonucleotide expression array.
2008-06-15 | E-GEOD-7850 | biostudies-arrayexpress