CXCR4/miR-1910-5p/MMRN2 Axis is Involved in Corneal Neovascularization by Affecting Vascular Permeability
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PROVIDER: PRJNA939144 | ENA |
REPOSITORIES: ENA
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Project description:To further investigate the molecular functions of CXCR4 in angiogenesis, we knocked out CXCR4 in HCE-T-cell lines, which were further undergo hypoxic treatment, using the CRISPR‒Cas9 system.
2023-04-26 | GSE226185 | GEO
Project description:PurposeChemokine receptor 4 (CXCR4) plays an essential role in the early stage of corneal neovascularization (CNV), but the underlying key molecular mechanism has yet to be addressed. This study aimed to explore the new molecular mechanism of CXCR4 in CNV and the related pathological events.MethodsCXCR4 was assayed by immunofluorescence or Western blotting. The function of the supernatant from hypoxia-treated human corneal epithelial cells (HCE-T) cells was examined by culturing with human umbilical vein endothelial cells. MicroRNA sequencing was used to detect the downstream microRNAs upon CXCR4 knockdown and analyzed by preliminary bioinformatics. The proangiogenic functions and downstream target genes of microRNA were investigated by gene interference and luciferase assay. An alkali-burned murine model was introduced to examine the function and mechanism of miR-1910-5p in vivo.ResultsHigh CXCR4 expression was confirmed in corneal tissues of patients with CNV and hypoxic HCE-T cells. The supernatant from hypoxia-treated HCE-T cells is involved in the CXCR4-mediated angiogenesis of human umbilical vein endothelial cells. Notably, miR-1910-5p was demonstrated to be at a high level in wild-type HCE-T cells and its supernatant, and in CNV patient tears. The proangiogenic functions of miR-1910-5p were demonstrated with the assays of cell migration, tube formation, and aortic ring. Moreover, miR-1910-5p significantly inhibited multimerin-2 expression by targeting its 3' untranslated region and caused significant extracellular junctional defects in human umbilical vein endothelial cells. MiR-1910-5p antagomir could significantly increase multimerin-2 level and decrease vascular leakage, and ultimately inhibit CNV in a murine model.ConclusionsOur results revealed a novel CXCR4-mediated mechanism and proved that targeting the miR-1910-5p/multimerin-2 pathway could be a promising therapeutic target for CNV.
| S-EPMC10108737 | biostudies-literature
Project description:In order to find out the vital genes during the corneal neovascularization (CNV), we set up two CNV models, one being induced by suture placement and the other by alkali burn to the corneal center. Young adult Balb/c and C57BL/6 were used. The corneas were checked daily for CNV development and some of the corneas were removed at two pre-determined time points for RNA extraction. Preliminary experiment indicated slightly different dynamics in suture- or chemical burn-induced CNV. Vigorous growth appeared at day 5 and day 6 in S-CNV and CB-CNV, and reached maximum length around day 10 and day 14, respectively, in S-CNV and CB-CNV. To check whether genetic MHC background has any effect on development of CNV, we compared CB-CNV in Balb/c and C57BL/6 mice at day 6. Corneas from Balb/c mice were isolated 5 and 10 days after suture-induced CNV, and 6 and 14 days after chemical burn-induced CNV. Corneas from C57BL/6 mice were isolated 6 days after chemical burn-induced CNV. Each time point had two or three biological replications.
2011-02-08 | E-GEOD-23347 | biostudies-arrayexpress
Project description:In order to find out the vital genes during the corneal neovascularization (CNV), we set up two CNV models, one being induced by suture placement and the other by alkali burn to the corneal center. Young adult Balb/c and C57BL/6 were used. The corneas were checked daily for CNV development and some of the corneas were removed at two pre-determined time points for RNA extraction. Preliminary experiment indicated slightly different dynamics in suture- or chemical burn-induced CNV. Vigorous growth appeared at day 5 and day 6 in S-CNV and CB-CNV, and reached maximum length around day 10 and day 14, respectively, in S-CNV and CB-CNV. To check whether genetic MHC background has any effect on development of CNV, we compared CB-CNV in Balb/c and C57BL/6 mice at day 6.
2011-02-08 | GSE23347 | GEO
Project description:Purpose Corneal neovascularization (CNV) impairs corneal transparency and visual acuity. The study aims to deepen our understanding of the molecules involved in CNV induced by alkali burns, facilitate a better grasp of CNV mechanisms, and uncover potential therapeutic targets. Methods Mice were selected for establishing CNV models via alkali burns. On days 3, 7, and 14 after the burns, corneal observations and histological investigations were conducted. An integrated analysis of RNA sequencing (RNA-seq)-based transcriptomics and label-free quantitative proteomics was performed in both normal and burned corneas. Bioinformatics approaches, encompassing GO and KEGG analysis, were applied to discern differentially expressed genes (DEGs) and crucial signaling pathways. Four potentially CNV-related genes were validated using qRT-PCR and Western blot. Results Significant CNV was observed on the seventh day. Forty-one genes were differentially expressed in neovascularized corneas, with 15 upregulated and 26 downregulated at both mRNA and protein levels. Bioinformatics analysis revealed that these DEGs participated in diverse biological processes, encompassing retinol and retinoic acid metabolism, neutrophil chemotaxis, and actin filament assembly, along with significant enrichment pathways like cytochrome P450, tyrosine, and phenylalanine metabolism. The upregulation of lymphocyte cytosolic protein 1 (LCP1) and cysteine and glycine-rich protein 2 (CSRP2) genes and the downregulation of transglutaminase 2 (TGM2) and transforming growth factor-beta-induced (TGFBI) genes were confirmed. Conclusions We analyzed gene expression differences in mouse corneas seven days after alkali burns, finding 41 genes with altered expression. The exact role of these genes in CNV is not fully understood, but exploring angiogenesis-related molecules offers potential for CNV treatment or prevention.
2024-01-26 | PXD047367 | Pride
Project description:The goal of this study was to identify signalling molecules downstream of CXCR4 in breast cancer cells. For this purpose, we sorted CXCR4-positive and CXCR4-negative cells from MDA-MB-231 breast cancer cell line by flow cytometry and performed microarrays analysis. Three sets of samples, each in quadruplicate, were analyzed. These include CXCR4-positive subpopulation, CXCR4-positive subpopulation treated with SDF-1 (ligand for CXCR4, also called CXCL12) for one hour and CXCR4-negative subpopulation.
2010-04-28 | E-GEOD-15893 | biostudies-arrayexpress
Project description:Fuchs endothelial corneal dystrophy (FECD) is a vision impairing pathology affecting the endothelial cells of the cornea. To better understand the disease, we developed a method to cultivate FECD cells isolated from surgical specimens. Using gene profiling, we compared the mRNA profiles of passage 2 FECD cells with passage 2 non-pathological corneal endothelial cells isolated from eye bank donor corneas.
2018-03-20 | GSE112039 | GEO
Project description:The goal of this study was to identify signaling molecules downstream of CXCR4 in breast cancer cells. For this purpose, we sorted CXCR4-positive and CXCR4-negative cells from MDA-MB-231 breast cancer cell line by flow cytometry and performed microarrays analysis.
2010-04-20 | GSE15893 | GEO
Project description:Antibodies and derivative drugs targeting immune checkpoints have been approved for the treatment of several malignancies, but there are fewer responses in patients with pancreatic cancer. Here, we designed a nanobody molecule with bi-targeting on PD-L1 and CXCR4, as both targets are overexpressed in many cancer cells and play important roles in tumorigenesis. The nanobody sequences targeting PD-L1 and CXCR4 were linked by the (G4S)3 flexible peptide to construct the anti-PD-L1/CXCR4 bispecific nanobody. The bispecific nanobody was expressed in E. coli cells and purified by affinity chromatography. The purified nanobody was biochemically characterized by mass spectrometry, Western blotting and flow cytometry to confirm the molecule and its association with both PD-L1 and CXCR4. The biological function of the nanobody and its anti-tumour effects were examined.
2023-03-11 | PXD037102 | Pride
Project description:Glucocorticoid Receptor Signaling is Critical for Mouse Corneal Development, Inhibition of Inflammatory Response and Neovascularization of the Cornea
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