Similar Datasets
Project description:To identify genes induced by Norrin in bEnd.3 cells (mouse brain endothelial cell line), we treated the cells with recombinant Norrin. It is known that the Norrin ligand induces beta-catenin-dependent (also known as canonical) signaling through binding to a receptor complex containing Frizzled4 (FZD4). First, we confirmed that receptor complex components, FZD4, LRP5/6, and TSPAN12, were expressed in bEnd.3 cells. Next, we observed that Norrin treatment strongly induces Axin2, a beta-catenin-dependent signaling response gene. Multiple norrin-induced DEGs were identified, including P2ry1. However, it is noted that many DEGs identified in previous scRNAseq studies on retinal vascular cells in Ndp and Tspan12 KO mice are not induced in bEnd.3. cells. This observation suggests that bEnd.3 cells are a good model system for studying signal initiation and signal transduction; however, they may not capture all aspects of cellular behavior.
2026-04-11 | GSE299490 | GEO
Project description:Blood-retina barrier (BRB) formation and retinal angiogenesis depend on beta-catenin signaling induced by the ligand norrin (NDP), the receptor frizzled4 (FZD4), co-receptor LRP5, and the tetraspanin TSPAN12. Impaired NDP/FZD4 signaling causes familial exudative vitreoretinopathy (FEVR), which may lead to blindness. Endothelial-cell specific inactivation of the Tspan12 gene at P28 using a Cdh5-CreERT2 driver shows that TSPAN12 functions in ECs to promote vascular morphogenesis and BRB formation in developing mice, and BRB maintenance in adult mice. 12 month after Tspan12 inactivation and loss of BRB maintenance with massive IgG and albumin extravasation we observe complement activation, cystoid edema, and impaired beta-wave in electroretinograms. RNA-Seq 6 month after Tspan12 inactivation provides a detailed view on the transcriptional response, including activation of antibody effector systems (complement and Fc receptors), inflammation and microglia responses, extracellular matrix organization and remodeling, and other responses.
2018-10-29 | GSE113878 | GEO
Project description:Frizzled4 (FZD4) is a pivotal receptor for Norrin and Wnt7a/b that mediates CNS angiogenesis, blood-brain barrier function and blood-retina barrier function. FZD4 is a target for pharmacological interventions in retinal and neurological disease. In order to understand mechanisms of regulation of Frizzled4 activity, two proximity biotinylation screens were performed, one in HEK293T cells and one in HeLa cells. For the screen in HEK293T cells, cells were transiently transfected. 24 hours later, cells were stimulated with 50 µM biotin for an additional 24 hours. Samples were processed and subjected to LC-MS/MSas described (PMID: 29516480). Samples S480 and S483 were biological replicates of cells co-transfected with V5-FZD4-BioID, HA-LRP5, and HA-TSPAN12 to reconstitute the norrin receptor complex. Samples S481 and S484 were biological replicates of cells co-transfected with V5-FZD4-BioID, HA-LRP5, and HA-TSPAN12 that were stimulated with flag-AP-Norrin conditioned media during the 24 hour biotin incubation period to induce norrin/frizzled4 signaling. Sample S479 and S482 were biological replicates of cells co-transfected with GFP-BioID, HA-LRP5, and HA-TSPAN12 as negative control. For the screen in HeLa cells, cells were transfected and biotinylated as described above. Sample S454 was transfected with V5-FZD4 (without BioID fusion, as a negative control). Sample S455 was transfected with GFP-BioID as another negative control. Sample S456 and S457 were transfected with V5-FZD4-BioID, of which only S457 was stimulated with flag-AP-Norrin conditioned medium during the biotin incubation period in order to induce FZD4 endocytosis. Multiple known and novel proximity interactors were identified in HEK293T and HeLa cells with high specificity. Comparisons of cells stimulated with norrin vs. vehicle indicate that proximity interactors of FZD4 were largely similar in cells with and without norrin stimulation in an overexpression context.
2026-05-15 | PXD065058 | Pride
Project description:RNA-seq analysis of NDP and F4L5.13 treated bEnd.3 endothelial cell line
| PRJNA701921 | ENA
Project description:Frizzled4 (FZD4) is a pivotal molecule in the CNS vasculature and a target for therapeutic intervention. Little is known how FZD4 is regulated. A proximity biotinylation screen with FZD4 as bait in a mouse brain endothelial cell line was performed. The goal is to identify candidate regulators of FZD4.
2026-04-14 | PXD077093 | Pride
Project description:Identifying Norrin-induced genes in bEnd.3 cells
| PRJNA1274721 | ENA
Project description:Medulloblastoma (MB), a tumor of the cerebellum, is the most common malignant brain tumor in children. One third of all human MB exhibits a gene expression signature of Sonic hedgehog (Shh) signaling. Hedgehog (Hh) pathway inhibitors have shown efficacy in clinical trials for MB, however, tumors develop resistance to these compounds, highlighting the need to identify additional therapeutic targets for treatment. We have identified a role for Norrin signaling in tumor initiation in the Patched (Ptch) mouse model of MB. Norrin is a secreted factor that functions as an atypical Wnt by binding to the Frizzled4 (Fzd4) receptor on endothelial cells to activate canonical beta-catenin-mediated Wnt signaling pathway. In the cerebellum, activation of Norrin/Fzd4 signaling is required for the establishment and maintenance of the blood brain barrier (BBB). We have identified a role for Norrin signaling in the stroma as a potent tumor inhibitory signal. Inactivation of Norrin in Ptch+/- mice significantly shortens latency and increases MB incidence. This phenotype is associated with an increased frequency of pre-tumor lesions and their conversion to malignancy. In this context, loss of Norrin signalling in endothelial cells is associated with an accelerated transition to a pro-tumor stroma characterized by vascular permeability, inflammation and angiogenic remodelling. Accordingly, loss of Ndp significantly alters the stromal gene expression signature of established Ptch MB.
2016-10-12 | GSE81962 | GEO
Project description:Medulloblastoma (MB), a tumor of the cerebellum, is the most common malignant brain tumor in children. One third of all human MB exhibits a gene expression signature of Sonic hedgehog (Shh) signaling. Hedgehog (Hh) pathway inhibitors have shown efficacy in clinical trials for MB, however, tumors develop resistance to these compounds, highlighting the need to identify additional therapeutic targets for treatment. We have identified a role for Norrin signaling in tumor initiation in the Patched (Ptch) mouse model of MB. Norrin is a secreted factor that functions as an atypical Wnt by binding to the Frizzled4 (Fzd4) receptor on endothelial cells to activate canonical beta-catenin-mediated Wnt signaling pathway. In the cerebellum, activation of Norrin/Fzd4 signaling is required for the establishment and maintenance of the blood brain barrier (BBB). We have identified a role for Norrin signaling in the stroma as a potent tumor inhibitory signal. Inactivation of Norrin in Ptch+/- mice significantly shortens latency and increases MB incidence. This phenotype is associated with an increased frequency of pre-tumor lesions and their conversion to malignancy. In this context, loss of Norrin signalling in endothelial cells is associated with an accelerated transition to a pro-tumor stroma characterized by vascular permeability, inflammation and angiogenic remodelling. Accordingly, loss of Ndp significantly alters the stromal gene expression signature of established Ptch MB.
2016-10-12 | GSE81963 | GEO
Project description:To characterize the long-term effect on the transcriptome of a decrement in Norrin/Fz4/Lrp signaling, microarray hybridization was performed with RNA from acutely dissociated and anti-PECAM immunoaffinity-purified adult WT, Fz4-/-, Lrp5-/-, and Norrin- retinal vascular cells.
2009-10-01 | GSE16703 | GEO
Project description:Mucin-domain glycoproteins populate the brain endothelial cell surface, but their roles as a class of glycoconjugates are largely unknown. Here, we perform bulk RNA-sequencing of bEnd.3 cells treated with StcE, which enzymatically degrades cell surface mucins, in order to understand the effects of mucin removal on brain endothelial cell function.
2024-12-17 | GSE268263 | GEO