GO:1900747 negative regulation of vascular endothelial growth factor signaling pathway: Mechanism, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1900747 describes any biological process that stops, prevents, or reduces the frequency, rate, or extent of vascular endothelial growth factor (VEGF) signaling.
• VEGF signaling is initiated by VEGF-A binding to receptors such as VEGFR2 (KDR) and co-receptors like neuropilin-1 (NRP1), triggering downstream MAPK/ERK and other cascades.
• Negative regulation of VEGF signaling is critical for balancing angiogenesis, vascular permeability, and immune cell infiltration in tumors.
• Key negative regulators include soluble VEGFR1 (sFlt-1), miR-200b, and glycoRNA-heparan sulfate complexes that modulate VEGF-A availability and receptor activation.
• Dysregulation of this process contributes to cancer progression, particularly triple-negative breast cancer stemness and metastasis, as well as vascular pathologies.
• CRISPR-based knockout, point mutation, knock-in, and overexpression models enable precise dissection of negative regulators in VEGF signaling for drug discovery.
Description
The vascular endothelial growth factor (VEGF) signaling pathway is a central regulator of angiogenesis, vascular permeability, and endothelial cell survival. Under physiological conditions, VEGF signaling must be tightly controlled to prevent excessive or insufficient vessel formation. The Gene Ontology term GO:1900747, negative regulation of vascular endothelial growth factor signaling pathway, encompasses any process that stops, prevents, or reduces the frequency, rate, or extent of VEGF signaling. This term is essential for understanding how cells maintain vascular homeostasis and how disruptions contribute to diseases such as cancer, retinopathies, and inflammatory disorders. Research has identified multiple layers of negative regulation, including soluble decoy receptors like soluble VEGFR1 (sFlt-1), microRNAs such as miR-200b, and extracellular matrix components that sequester VEGF-A. For example, glycoRNA complexed with heparan sulfate has been shown to regulate VEGF-A signaling, highlighting a novel extracellular mechanism. In tumors, negative regulation of VEGF signaling influences immune cell infiltration and the formation of high endothelial venules (HEVs), which are associated with improved responses to immunotherapy. Understanding GO:1900747 is therefore critical for developing therapeutic strategies that modulate angiogenesis. This article synthesizes current knowledge on the mechanisms, key genes, disease relevance, and research methods for studying negative regulation of VEGF signaling, with a focus on CRISPR-based models for functional validation.
negative regulation of vascular endothelial growth factor signaling pathway At A Glance
| GO ID | GO:1900747 |
|---|---|
| GO term | negative regulation of vascular endothelial growth factor signaling pathway |
| Ontology | biological_process |
| Synonym | inhibition of VEGF signaling; downregulation of VEGF-activated signaling pathway; negative regulation of VEGF signaling |
| Major function | Suppresses VEGF-induced angiogenesis, vascular permeability, and endothelial cell proliferation |
| Related pathways | VEGFR2 signaling, MAPK/ERK cascade, Notch signaling, heparan sulfate-glycoRNA interactions |
| Key regulators | sFlt-1, miR-200b, NRP1, GAPVD1, glycoRNA-heparan sulfate complexes |
| Disease relevance | Cancer (triple-negative breast cancer), vascular permeability disorders, immunotherapy response |
What Is GO:1900747?
GO:1900747 is a biological process term defined as any process that stops, prevents, or reduces the frequency, rate, or extent of the vascular endothelial growth factor signaling pathway. In simpler terms, it covers all molecular and cellular events that put the brakes on VEGF-driven signals, thereby controlling blood vessel growth and vascular function.
Why Is negative regulation of vascular endothelial growth factor signaling pathway Important in Cell Biology?
Negative regulation of VEGF signaling is vital for preventing pathological angiogenesis and maintaining vascular quiescence. Dysregulation of this process is implicated in tumor progression, where excessive VEGF signaling promotes cancer stemness and immune evasion, while insufficient negative regulation can lead to vascular leakage and edema. Understanding GO:1900747 provides insights into therapeutic strategies that aim to normalize tumor vasculature and enhance immunotherapy efficacy.
• Controls angiogenesis to prevent excessive blood vessel formation in tumors and retinopathies.
• Regulates vascular permeability by modulating endothelial adherens junctions through NRP1.
• Influences cancer stemness in triple-negative breast cancer via MAPK/ERK pathway.
• Promotes formation of high endothelial venules (HEVs) that enhance anti-tumor immunity when combined with anti-PD-L1 therapy.
• Involves extracellular mechanisms such as glycoRNA-heparan sulfate complexes that sequester VEGF-A.
• MicroRNAs like miR-200b directly target VEGF signaling components to fine-tune pathway activity.
• Soluble VEGFR1 (sFlt-1) acts as a decoy receptor to negatively regulate VEGF signaling.
• Notch signaling interacts with VEGF to regulate endothelial cell differentiation and arterial specification.
• Dysregulation contributes to cancer progression, making it a target for anti-angiogenic therapies.
• CRISPR screens can identify novel negative regulators, accelerating drug target discovery.
What Happens During negative regulation of vascular endothelial growth factor signaling pathway?
Sequestration of VEGF Ligand
In simple terms: Molecules trap VEGF so it cannot reach its receptors.
Negative regulation can occur extracellularly by sequestering VEGF-A. For instance, glycoRNA complexed with heparan sulfate binds VEGF-A and prevents it from activating VEGFR2, thereby dampening downstream signaling. Soluble VEGFR1 (sFlt-1) acts as a decoy receptor that binds VEGF-A with high affinity, reducing its availability to VEGFR2. This mechanism is crucial for maintaining vascular homeostasis.
Receptor-Level Inhibition
In simple terms: The activity of VEGF receptors is blocked or reduced.
Negative regulation can target VEGFR2 directly. Neuropilin-1 (NRP1) controls vascular permeability through juxtacrine regulation of endothelial adherens junctions, which can modulate VEGFR2 signaling. Additionally, microRNA miR-200b has been shown to downregulate VEGF signaling by targeting components of the pathway. These receptor-level events reduce the frequency and extent of VEGF signal transduction.
Downstream Signaling Attenuation
In simple terms: Signals inside the cell that promote growth are turned down.
VEGF signaling activates MAPK/ERK cascades that promote cell proliferation and stemness. Negative regulation can occur through phosphatases or inhibitory proteins that dephosphorylate key intermediates. For example, the VEGFA/NRP-1/GAPVD1 axis enhances tumor cell-macrophage crosstalk and cancer stemness, and its inhibition could represent a negative regulatory mechanism. Notch signaling also intersects with VEGF to regulate endothelial differentiation, providing another layer of negative control.
Transcriptional and Post-Transcriptional Control
In simple terms: Cells reduce the production of VEGF pathway components.
Negative regulation can be achieved by reducing the expression of VEGF or its receptors. MicroRNAs such as miR-200b directly target VEGF signaling molecules, leading to decreased pathway activity. Transcriptional repressors may also downregulate VEGFR2 expression. These mechanisms ensure that VEGF signaling is kept in check under normal conditions.
Key Genes Involved in GO:1900747 negative regulation of vascular endothelial growth factor signaling pathway
The following genes and proteins are key players in the negative regulation of VEGF signaling, based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| VEGFA | Ligand that initiates signaling; its sequestration negatively regulates pathway | Target for glycoRNA-heparan sulfate and sFlt-1 decoy |
| KDR (VEGFR2) | Main receptor for VEGF; its inhibition reduces signaling | Key target for negative regulators and CRISPR knockout studies |
| FLT1 (VEGFR1) | Soluble form (sFlt-1) acts as decoy receptor | Negative regulator of VEGF availability |
| NRP1 | Co-receptor that modulates VEGFR2 signaling and permeability | Juxtacrine regulation of adherens junctions |
| GAPVD1 | Involved in VEGFA/NRP-1 axis promoting cancer stemness | Potential target for negative regulation in TNBC |
| miR-200b | MicroRNA that downregulates VEGF signaling | Post-transcriptional negative regulator |
| NOTCH1 | Interacts with VEGF to regulate endothelial differentiation | Negative crosstalk in arterial specification |
| DLL4 | Notch ligand that modulates VEGF-induced angiogenesis | Potential negative regulator of VEGF signaling |
| HIF1A | Transcription factor that induces VEGF under hypoxia | Its inhibition reduces VEGF signaling |
| PTPN11 (SHP2) | Phosphatase that can dephosphorylate VEGFR2 | Potential negative regulator of downstream signaling |
| PTEN | Lipid phosphatase that antagonizes PI3K/AKT downstream of VEGF | Negative regulator of survival signals |
| SPRY2 | Sprouty protein that inhibits MAPK/ERK cascade | Negative feedback regulator of VEGF signaling |
| DUSP1 | Dual-specificity phosphatase that inactivates ERK | Attenuates VEGF-induced MAPK signaling |
| CDKN1B (p27) | Cell cycle inhibitor that can be upregulated by negative regulators | Reduces endothelial proliferation |
| THBS1 | Thrombospondin-1, an endogenous angiogenesis inhibitor | Negative regulator of VEGF signaling |
| SERPINF1 | Pigment epithelium-derived factor, anti-angiogenic | Inhibits VEGF-induced angiogenesis |
| VASH1 | Vasohibin-1, negative feedback regulator of angiogenesis | Inhibits VEGF signaling in endothelial cells |
| miR-126 | MicroRNA that modulates VEGF signaling | Potential negative regulator in endothelial cells |
How Is negative regulation of vascular endothelial growth factor signaling pathway Regulated?
Negative regulation of VEGF signaling is itself tightly controlled. Hypoxia induces HIF1A, which upregulates VEGF but also triggers negative feedback loops involving VASH1 and other inhibitors. Notch signaling, activated by DLL4, can suppress VEGF-induced tip cell formation, thereby negatively regulating angiogenesis. Additionally, microRNAs such as miR-200b are transcriptionally regulated by developmental cues and stress signals. Extracellular matrix components like heparan sulfate modulate the bioavailability of VEGF-A and its interaction with glycoRNA, adding another layer of regulation.
negative regulation of vascular endothelial growth factor signaling pathway and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| GAPVD1 | Triple-negative breast cancer stemness | Knockout in TNBC cell lines (e.g., MDA-MB-231) |
| NRP1 | Vascular permeability disorders | Endothelial cell-specific knockout in mice |
| VEGFA | Angiogenesis-related retinopathies | Knock-in of soluble decoy receptor in zebrafish |
| FLT1 | Preeclampsia and cancer | Overexpression of sFlt-1 in placental cells |
| miR-200b | Cancer progression and metastasis | Overexpression in cancer cell lines |
Triple-Negative Breast Cancer
In triple-negative breast cancer (TNBC), VEGF signaling promotes cancer stemness and progression via the MAPK/ERK pathway. The VEGFA/NRP-1/GAPVD1 axis enhances tumor cell-macrophage crosstalk, contributing to stemness and metastasis. Negative regulation of VEGF signaling could therefore reduce cancer stem cell populations and inhibit tumor growth. Targeting this pathway with CRISPR knockout of GAPVD1 or NRP1 may provide therapeutic benefits.
Tumor Immunity and High Endothelial Venules
Combined antiangiogenic therapy and anti-PD-L1 blockade stimulates tumor immunity through the formation of high endothelial venules (HEVs). Negative regulation of VEGF signaling is thought to promote HEV formation, which facilitates lymphocyte infiltration into tumors. This highlights the importance of balancing VEGF signaling to enhance immunotherapy responses.
Vascular Permeability Disorders
Neuropilin-1 controls vascular permeability through juxtacrine regulation of endothelial adherens junctions. Dysregulation of NRP1-mediated negative regulation can lead to vascular leakage, edema, and inflammation. Understanding these mechanisms is relevant for diseases such as sepsis and diabetic retinopathy.
From negative regulation of vascular endothelial growth factor signaling pathway-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of GAPVD1 reduce VEGF-induced cancer stemness? | CRISPR knockout of GAPVD1 in TNBC cells |
| Can a point mutation in VEGFR2 prevent negative regulation? | CRISPR point mutation at phosphorylation sites in KDR |
| Does overexpression of sFlt-1 inhibit tumor angiogenesis? | Knock-in of soluble FLT1 in mouse models |
| How does NRP1 glycosylation affect VEGF signaling? | Point mutation of NRP1 glycosylation sites |
| What is the role of miR-200b in VEGF signaling? | Overexpression or knockout of miR-200b in endothelial cells |
| Can tagged VEGFR2 track receptor trafficking? | Knock-in of fluorescent tag into KDR locus |
How to Study the negative regulation of vascular endothelial growth factor signaling pathway Process
| Method | What It Measures | Typical Application |
|---|---|---|
| CRISPR knockout screen | Loss-of-function effects on VEGF signaling | Identify novel negative regulators |
| RNA-seq | Transcriptional changes | Measure miR-200b target downregulation |
| Phosphoproteomics | Phosphorylation of VEGFR2 and downstream targets | Quantify pathway inhibition |
| Immunofluorescence | Protein localization and adherens junction integrity | Study NRP1 function |
| Tube formation assay | Angiogenic capacity in vitro | Test negative regulators |
| Vascular permeability assay | Endothelial barrier function | Assess NRP1 regulation |
| Western blot | Protein expression and phosphorylation | Validate CRISPR knockouts |
| qRT-PCR | mRNA levels of VEGF pathway genes | Confirm overexpression or knockout |
CRISPR Knockout Screens
Genome-wide CRISPR knockout screens can identify novel negative regulators of VEGF signaling. For example, knocking out candidate genes in endothelial cells followed by VEGF stimulation and phospho-ERK readout can reveal inhibitors. This approach is unbiased and scalable.
RNA Sequencing and Transcriptomics
RNA-seq can measure changes in gene expression upon modulation of negative regulators. For instance, overexpression of miR-200b leads to downregulation of VEGF pathway genes, which can be quantified by RNA-seq. This method provides a global view of transcriptional changes.
Proteomics and Phosphoproteomics
Mass spectrometry-based phosphoproteomics can quantify changes in VEGFR2 phosphorylation and downstream signaling upon negative regulation. This is useful for identifying specific phosphorylation sites affected by inhibitors.
Imaging and Functional Assays
Live-cell imaging of endothelial tube formation and vascular permeability assays can assess the functional impact of negative regulators. For example, NRP1-mediated adherens junction regulation can be visualized using immunofluorescence.
How CRISPR Can Be Used to Study GO:1900747 negative regulation of vascular endothelial growth factor signaling pathway
Knockout
CRISPR knockout of negative regulators such as GAPVD1 or NRP1 can reveal their role in VEGF signaling. For example, knocking out GAPVD1 in TNBC cells may reduce cancer stemness and tumor growth. Knockout of miR-200b could increase VEGF signaling, confirming its negative regulatory function.
Point Mutation
Introducing point mutations in VEGFR2 phosphorylation sites or NRP1 glycosylation sites can dissect their role in negative regulation. For instance, mutating specific tyrosines in KDR can prevent receptor internalization, affecting signaling duration.
Knock-in
Knock-in of tagged VEGFR2 or sFlt-1 can enable live-cell imaging and tracking of receptor dynamics. Knock-in of a fluorescent tag into the KDR locus allows visualization of receptor trafficking under negative regulation.
Overexpression
Overexpression of negative regulators like sFlt-1 or miR-200b can suppress VEGF signaling and angiogenesis. This approach is useful for validating therapeutic targets and testing combination with anti-PD-L1 therapy.
How EDITGENE Supports negative regulation of vascular endothelial growth factor signaling pathway Research
Researchers studying negative regulation of vascular endothelial growth factor signaling pathway-related genes often need to determine whether a candidate gene is causally involved in suppressing VEGF-driven phenotypes. EDITGENE provides a comprehensive suite of CRISPR services to enable precise genetic manipulation and functional validation.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of vascular endothelial growth factor signaling pathway research.
Frequently Asked Questions About negative regulation of vascular endothelial growth factor signaling pathway
What is GO:1900747?
GO:1900747 is a Gene Ontology biological process term for any process that stops, prevents, or reduces the frequency, rate, or extent of vascular endothelial growth factor signaling pathway.
What genes are involved in negative regulation of VEGF signaling?
Key genes include VEGFA, KDR (VEGFR2), FLT1 (sFlt-1), NRP1, GAPVD1, miR-200b, and NOTCH1, among others.
How does negative regulation of VEGF signaling affect cancer?
It can suppress tumor angiogenesis and cancer stemness; for example, the VEGFA/NRP-1/GAPVD1 axis promotes TNBC progression, and its inhibition may reduce stemness.
What is the role of miR-200b in VEGF signaling?
miR-200b negatively regulates VEGF signaling by targeting components of the pathway, reducing downstream MAPK/ERK activation.
How can CRISPR be used to study negative regulation of VEGF signaling?
CRISPR knockout, point mutation, knock-in, and overexpression models allow functional dissection of candidate negative regulators in endothelial and cancer cells.
What diseases are associated with dysregulated VEGF signaling?
Cancer, vascular permeability disorders, retinopathies, and preeclampsia are linked to altered negative regulation of VEGF signaling.
What is the role of neuropilin-1 in VEGF signaling?
NRP1 acts as a co-receptor that modulates VEGFR2 signaling and controls vascular permeability through adherens junctions.
How does glycoRNA regulate VEGF-A signaling?
GlycoRNA complexed with heparan sulfate binds VEGF-A and negatively regulates its signaling, affecting angiogenesis.
What are high endothelial venules (HEVs) and their link to VEGF?
HEVs are specialized blood vessels that support lymphocyte infiltration; negative regulation of VEGF signaling promotes HEV formation and enhances anti-tumor immunity.
What methods are used to study negative regulation of VEGF signaling?
CRISPR screens, RNA-seq, phosphoproteomics, imaging, and functional assays such as tube formation and permeability assays are commonly used.
Conclusion
GO:1900747, negative regulation of vascular endothelial growth factor signaling pathway, is a critical biological process that maintains vascular homeostasis and prevents pathological angiogenesis. Key regulators such as sFlt-1, miR-200b, NRP1, and glycoRNA-heparan sulfate complexes modulate VEGF signaling at multiple levels. Dysregulation of this process contributes to cancer progression, particularly in triple-negative breast cancer, and influences tumor immunity. CRISPR-based models are indispensable for dissecting these mechanisms and identifying new therapeutic targets. EDITGENE provides comprehensive CRISPR services to support research in this field.
References
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- 3. Pal S et al.. 2024. Neuropilin-1 controls vascular permeability through juxtacrine regulation of endothelial adherens junctions.. Angiogenesis 28(1):7 PMID: 39668325
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