GO:0090051 negative regulation of cell migration involved in sprouting angiogenesis: Mechanism, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0090051 describes any process that decreases the frequency, rate or extent of endothelial cell migration during sprouting angiogenesis, a key step in new blood vessel formation.
• Negative regulators such as Nogo-A and EPAC2 restrict endothelial cell migration and tube formation, preventing excessive or disorganized angiogenesis.
• The p38 MAPK pathway is required for mural cell recruitment and vessel stabilization, indirectly limiting sprouting migration.
• DLL4/Notch signaling acts as a negative feedback mechanism that suppresses endothelial sprouting and migration under hypoxic conditions.
• Dysregulation of this process contributes to diseases including CNS angiogenesis disorders, abdominal aortic aneurysm, and cancer progression.
• CRISPR-based knockout, knock-in, and overexpression models enable causal interrogation of genes that negatively regulate sprouting angiogenesis.
Description
Sprouting angiogenesis is the process by which new blood vessels emerge from pre-existing vasculature, driven by the orderly migration of endothelial cells into the extracellular matrix. This migratory step is tightly controlled by positive and negative signals to ensure proper vessel patterning. GO:0090051, negative regulation of cell migration involved in sprouting angiogenesis, captures the biological processes that decrease the frequency, rate, or extent of this endothelial cell migration. Understanding this term is critical because excessive or insufficient sprouting migration underlies numerous pathological conditions, including tumor angiogenesis, ischemic disease, and vascular malformations. Research has identified several negative regulators that operate at different levels of the sprouting cascade. For example, Nogo-A inhibits CNS angiogenesis by restricting endothelial cell migration and sprout formation. EPAC2 acts as a negative regulator in Matrigel-driven tubulogenesis of human microvascular endothelial cells. The p38 MAPK pathway is required for mural cell recruitment, which stabilizes vessels and limits further sprouting. DLL4/Notch signaling suppresses endothelial cell migration and tube formation under hypoxia, providing a negative feedback loop. These findings highlight the diversity of molecular mechanisms that converge on GO:0090051. For researchers, GO:0090051 provides a framework to study how specific genes and pathways restrain angiogenesis. Dysregulation of these negative regulators is implicated in abdominal aortic aneurysm, bladder cancer, triple-negative breast cancer, and hepatic fibrosis. CRISPR-based functional genomics now allows systematic interrogation of these genes in endothelial and disease models, accelerating the discovery of therapeutic targets.
negative regulation of cell migration involved in sprouting angiogenesis At A Glance
| GO ID | GO:0090051 |
|---|---|
| GO term | negative regulation of cell migration involved in sprouting angiogenesis |
| Ontology | biological_process |
| Synonym | none |
| Major function | Decreases the frequency, rate or extent of endothelial cell migration during sprouting angiogenesis |
| Key negative regulators | Nogo-A, EPAC2, DLL4/Notch, p38 MAPK pathway components |
| Associated diseases | CNS angiogenesis disorders, abdominal aortic aneurysm, bladder cancer, triple-negative breast cancer |
| Research models | Endothelial cell migration assays, Matrigel tubulogenesis, CRISPR knockout/knock-in |
What Is GO:0090051?
GO:0090051 is a biological process term defined as any process that decreases the frequency, rate or extent of cell migration involved in sprouting angiogenesis. Cell migration involved in sprouting angiogenesis is the orderly movement of endothelial cells into the extracellular matrix in order to form new blood vessels contributing to the process of sprouting angiogenesis. In simpler terms, it covers all molecular and cellular events that put the brakes on endothelial cell movement during the formation of new vessel sprouts.
Why Is negative regulation of cell migration involved in sprouting angiogenesis Important in Cell Biology?
GO:0090051 is important because it defines the braking mechanisms that prevent excessive or disorganized sprouting angiogenesis. Without proper negative regulation, endothelial cells migrate uncontrollably, leading to vascular malformations, tumor progression, and inflammatory diseases. Conversely, excessive negative regulation can impair tissue repair and ischemia-induced neovascularization. Understanding this term helps researchers identify therapeutic targets for modulating angiogenesis in cancer, cardiovascular disease, and fibrosis.
• Controls endothelial cell migration to ensure proper vessel patterning during development and repair.
• Prevents excessive angiogenesis in tumors, where uncontrolled sprouting supports cancer growth.
• Dysregulation is linked to abdominal aortic aneurysm and vascular instability.
• Nogo-A-mediated negative regulation is critical for CNS angiogenesis and blood-brain barrier function.
• EPAC2 restricts tubulogenesis in human microvascular endothelial cells, highlighting its role in vascular homeostasis.
• p38 MAPK-dependent mural cell recruitment stabilizes vessels and limits further sprouting.
• DLL4/Notch signaling acts as a negative feedback loop under hypoxia, controlling choroid-retinal endothelial migration.
• Hepatic stellate cell activation and liver fibrosis involve angiogenic processes that may be modulated by negative regulators.
• CRISPR screening can identify novel negative regulators of sprouting angiogenesis for therapeutic targeting.
• Provides a conceptual framework for distinguishing pro- and anti-angiogenic signals in disease models.
What Happens During negative regulation of cell migration involved in sprouting angiogenesis?
Initiation of negative regulation by extracellular cues
In simple terms: External signals tell endothelial cells to slow down their movement.
Negative regulation of sprouting angiogenesis begins when extracellular cues, such as Nogo-A or DLL4, engage receptors on endothelial cells. Nogo-A acts as a negative regulator of CNS angiogenesis by inhibiting endothelial cell migration and sprout formation. DLL4/Notch signaling is activated under hypoxic conditions and suppresses endothelial cell migration and tube formation. These cues initiate intracellular signaling that counteracts pro-migratory pathways.
Intracellular signaling that restricts migration
In simple terms: Inside the cell, specific pathways put the brakes on the machinery that drives movement.
EPAC2 acts as a negative regulator in Matrigel-driven tubulogenesis of human microvascular endothelial cells, limiting the formation of tube-like structures. The p38 MAPK pathway is required for mural cell recruitment during angiogenesis in the rat aorta model, which stabilizes vessels and indirectly restricts further endothelial sprouting. These intracellular pathways modulate cytoskeletal dynamics and cell-matrix adhesion to reduce migration speed and directionality.
Cell-cell communication and mural cell recruitment
In simple terms: Support cells are recruited to stabilize vessels and stop new sprouts from forming.
Mural cells, such as pericytes and smooth muscle cells, are recruited to nascent vessels through p38 MAPK-dependent signaling. This recruitment provides physical and paracrine signals that stabilize the vessel and negatively regulate further endothelial cell migration. In the absence of proper mural cell coverage, vessels remain plastic and continue to sprout excessively.
Feedback inhibition and resolution of sprouting
In simple terms: Once enough vessels are formed, feedback loops shut down the sprouting process.
DLL4/Notch signaling provides a negative feedback loop that limits the number of endothelial tip cells and reduces migration under hypoxic conditions. This feedback ensures that sprouting is balanced and prevents excessive vessel density. Resolution of sprouting involves a shift from migratory to quiescent endothelial cell phenotypes, a process critical for vascular homeostasis.
Key Genes Involved in GO:0090051 negative regulation of cell migration involved in sprouting angiogenesis
The following genes and proteins have been experimentally implicated in the negative regulation of cell migration involved in sprouting angiogenesis, based on the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| RTN4 (Nogo-A) | Negative regulator of CNS angiogenesis; inhibits endothelial cell migration and sprout formation | Studied in CNS angiogenesis and blood-brain barrier models |
| RAPGEF4 (EPAC2) | Negative regulator in Matrigel-driven tubulogenesis of human microvascular endothelial cells | Target for modulating microvascular tube formation |
| DLL4 | Notch ligand; suppresses endothelial cell migration and tube formation under hypoxia | Key negative feedback regulator of sprouting angiogenesis |
| NOTCH1 | Receptor mediating DLL4-induced negative regulation of endothelial migration | Component of the DLL4/Notch signaling axis |
| MAPK14 (p38 MAPK) | Required for mural cell recruitment and vessel stabilization | Indirect negative regulator of sprouting migration |
| TRIM15 | Knockdown inhibits hepatic stellate cell activation, which is linked to angiogenic processes | Potential regulator of fibrosis-associated angiogenesis |
| MIR1-1 (miR-1-3p) | Circulating biomarker for abdominal aortic aneurysm; may influence vascular remodeling | Biomarker and potential regulator of vascular pathology |
| VEGFA | Pro-angiogenic factor; its activity is counterbalanced by negative regulators | Central to sprouting angiogenesis; target for negative regulation studies |
| KDR (VEGFR2) | Receptor for VEGFA; downstream signaling is modulated by negative regulators | Key node in pro- vs anti-migratory signaling |
| CDH5 (VE-cadherin) | Endothelial cell-cell adhesion molecule; affects migration and vessel stability | Marker of endothelial identity and junctional stability |
| ACTA2 (α-SMA) | Mural cell marker; recruited via p38 MAPK signaling | Indicator of mural cell recruitment and vessel stabilization |
| PDGFRB | Receptor for PDGF; involved in mural cell recruitment | Target for studying pericyte recruitment |
| HEY1 | Notch target gene; mediates DLL4-induced negative regulation | Readout of Notch activation in endothelial cells |
| HES1 | Notch target gene; suppresses endothelial sprouting | Marker of Notch-mediated negative feedback |
| CXCL8 (IL-8) | Pro-inflammatory chemokine; promotes pro-metastatic and angiogenic phenotypes | Link between inflammation and angiogenesis in TNBC |
| CCL2 (MCP-1) | Chemokine involved in tumor-stroma-inflammation networks | Modulates angiogenic and metastatic behavior |
| COL1A1 | Extracellular matrix component; affects endothelial cell migration | Matrix remodeling in tubulogenesis assays |
| FN1 (Fibronectin) | Extracellular matrix protein; supports endothelial migration | Substrate for Matrigel-driven tubulogenesis |
How Is negative regulation of cell migration involved in sprouting angiogenesis Regulated?
The negative regulation of cell migration involved in sprouting angiogenesis is controlled by multiple signaling pathways. Nogo-A inhibits CNS angiogenesis through receptor-mediated signaling that restricts endothelial cell motility. EPAC2 acts as a negative regulator in Matrigel-driven tubulogenesis, likely through cAMP-dependent mechanisms. The p38 MAPK pathway is required for mural cell recruitment, which stabilizes vessels and limits further sprouting. DLL4/Notch signaling provides a negative feedback loop that suppresses endothelial cell migration and tube formation under hypoxic conditions. Additionally, inflammatory networks involving chemokines such as CXCL8 and CCL2 can modulate angiogenic phenotypes in the tumor microenvironment. These pathways converge to balance pro- and anti-migratory signals during sprouting angiogenesis.
negative regulation of cell migration involved in sprouting angiogenesis and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| RTN4 (Nogo-A) | CNS angiogenesis disorders | Knockout mouse or endothelial cell-specific KO |
| DLL4 | Tumor angiogenesis, hypoxic retinal vascular disease | Endothelial cell overexpression or knockdown |
| RAPGEF4 (EPAC2) | Microvascular dysfunction | Knockout in human microvascular endothelial cells |
| TRIM15 | Liver fibrosis | Knockdown in hepatic stellate cells |
| MIR1-1 (miR-1-3p) | Abdominal aortic aneurysm | Circulating biomarker studies; overexpression in vascular cells |
Cancer and tumor angiogenesis
In tumors, excessive sprouting angiogenesis supports growth and metastasis. Negative regulators such as DLL4/Notch and Nogo-A can restrain endothelial cell migration, and their dysregulation contributes to tumor vascularization. In triple-negative breast cancer, tumor-stroma-inflammation networks promote pro-metastatic chemokines and aggressiveness, which may override negative regulation of sprouting angiogenesis. Krill oil has been shown to exhibit anti-tumor efficacy against bladder cancer by affecting tumor-associated angiogenic vasculature, highlighting the therapeutic potential of modulating angiogenesis.
Vascular and cardiovascular disease
Abdominal aortic aneurysm is associated with altered serum levels of miR-1-3p, a potential circulating biomarker, suggesting that microRNA-mediated regulation of vascular cells may influence disease progression. Impaired negative regulation of endothelial migration could contribute to vessel wall instability and aneurysm formation. p38 MAPK-dependent mural cell recruitment is critical for vessel stabilization, and its dysfunction may lead to vascular pathology.
CNS angiogenesis and neurological disorders
Nogo-A is a negative regulator of CNS angiogenesis, and its activity is important for maintaining blood-brain barrier integrity and proper vascular patterning in the central nervous system. Dysregulation of CNS angiogenesis is implicated in neurological disorders, including stroke and neurodegeneration. Understanding how Nogo-A and other negative regulators control endothelial migration may provide therapeutic opportunities for CNS vascular diseases.
Fibrosis and hepatic stellate cell activation
Knockdown of TRIM15 inhibits the activation of hepatic stellate cells, which are key drivers of liver fibrosis and associated angiogenesis. Although the direct link to sprouting angiogenesis is not fully established, hepatic stellate cell activation is often accompanied by pathological angiogenesis. Modulating negative regulators of endothelial migration may therefore influence fibrosis progression.
From negative regulation of cell migration involved in sprouting angiogenesis-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of Nogo-A increase endothelial cell migration in CNS angiogenesis? | RTN4 knockout in endothelial cells or mouse models |
| Does EPAC2 point mutation affect its negative regulation of tubulogenesis? | CRISPR point mutation knock-in in human microvascular endothelial cells |
| Can DLL4 overexpression suppress hypoxic endothelial migration? | DLL4 overexpression in choroid-retinal endothelial cells |
| Is p38 MAPK required for mural cell recruitment? | MAPK14 knockout in rat aorta model |
| Does TRIM15 knockdown inhibit hepatic stellate cell activation? | TRIM15 knockdown in hepatic stellate cells |
| Can miR-1-3p serve as a biomarker for abdominal aortic aneurysm? | Circulating miRNA profiling in patient serum |
How to Study the negative regulation of cell migration involved in sprouting angiogenesis Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Transwell migration assay | Number of migrated endothelial cells | Quantify negative regulation by Nogo-A or EPAC2 |
| Scratch wound assay | Rate of cell migration into a gap | Assess DLL4/Notch effects under hypoxia |
| Matrigel tube formation | Capillary-like network formation | Study EPAC2-mediated negative regulation |
| Endothelial spheroid sprouting | Number and length of sprouts | Model sprouting angiogenesis in 3D |
| CRISPR knockout | Loss-of-function phenotype | Test causal role of RTN4 in CNS angiogenesis |
| CRISPR knock-in | Point mutation or tag effects | Dissect EPAC2 domain function |
| RNA-seq | Transcriptomic changes | Identify downstream targets of negative regulators |
| Immunostaining | Protein localization and expression | Detect mural cell recruitment (α-SMA) |
Endothelial cell migration assays
Transwell and scratch wound assays are standard methods to measure endothelial cell migration. These assays can be used to quantify the effects of negative regulators such as Nogo-A or EPAC2 on cell motility. Hypoxia chambers can be incorporated to mimic pathological conditions, as shown for DLL4-mediated suppression of migration.
Tube formation and sprouting assays
Matrigel-driven tubulogenesis assays assess the ability of endothelial cells to form capillary-like structures, a surrogate for sprouting angiogenesis. EPAC2 was identified as a negative regulator using this assay. Three-dimensional sprouting assays from endothelial spheroids provide more physiologically relevant models.
CRISPR-based functional genomics
CRISPR knockout, knock-in, and overexpression models enable causal interrogation of genes involved in negative regulation of sprouting angiogenesis. For example, knockout of RTN4 can test its role in CNS angiogenesis, while point mutations in RAPGEF4 can dissect domain-specific functions. Pooled CRISPR screens can identify novel negative regulators in an unbiased manner.
Imaging and molecular readouts
Live-cell imaging of fluorescently tagged endothelial cells allows real-time tracking of migration and sprout dynamics. Immunostaining for markers such as VE-cadherin, α-SMA, and Notch targets (HEY1, HES1) provides molecular readouts of negative regulation. RNA-seq and proteomics can reveal downstream signaling changes.
How CRISPR Can Be Used to Study GO:0090051 negative regulation of cell migration involved in sprouting angiogenesis
Knockout
CRISPR knockout of negative regulators such as RTN4 or RAPGEF4 can test whether their loss increases endothelial cell migration and sprouting. For example, RTN4 knockout in endothelial cells would be expected to enhance CNS angiogenesis. Similarly, RAPGEF4 knockout in human microvascular endothelial cells can validate its role in restricting tubulogenesis.
Point Mutation
Point mutations can dissect specific domains or phosphorylation sites in negative regulators. For instance, mutating the catalytic domain of EPAC2 may abolish its negative regulation of tubulogenesis. CRISPR prime editing or homology-directed repair can introduce precise mutations to study structure-function relationships.
Knock-in
Knock-in of fluorescent tags or epitope tags allows real-time tracking of negative regulators in endothelial cells. Tagging endogenous DLL4 or Nogo-A can reveal their dynamic localization during sprouting. Knock-in of reporter genes under the control of target gene promoters can monitor transcriptional activity.
Overexpression
Overexpression of negative regulators such as DLL4 or Nogo-A can suppress endothelial cell migration and tube formation. DLL4 overexpression under hypoxic conditions inhibits choroid-retinal endothelial cell migration. Overexpression models are useful for gain-of-function studies and for testing therapeutic potential.
How EDITGENE Supports negative regulation of cell migration involved in sprouting angiogenesis Research
Researchers studying negative regulation of cell migration involved in sprouting angiogenesis-related genes often need to determine whether a candidate gene is causally involved in restricting endothelial cell migration or whether its effect is secondary to other pathways. EDITGENE provides CRISPR-based cell model services to enable precise genetic interrogation of these questions.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of cell migration involved in sprouting angiogenesis research.
Frequently Asked Questions About negative regulation of cell migration involved in sprouting angiogenesis
What is GO:0090051?
GO:0090051 is a Gene Ontology biological process term for negative regulation of cell migration involved in sprouting angiogenesis, defined as any process that decreases the frequency, rate or extent of endothelial cell migration during sprout formation.
What genes are involved in negative regulation of sprouting angiogenesis?
Key genes include RTN4 (Nogo-A), RAPGEF4 (EPAC2), DLL4, NOTCH1, and MAPK14 (p38 MAPK), among others.
How does Nogo-A inhibit angiogenesis?
Nogo-A acts as a negative regulator of CNS angiogenesis by inhibiting endothelial cell migration and sprout formation.
What is the role of EPAC2 in tubulogenesis?
EPAC2 acts as a negative regulator in Matrigel-driven tubulogenesis of human microvascular endothelial cells.
How does DLL4/Notch signaling affect endothelial migration?
DLL4/Notch signaling suppresses endothelial cell migration and tube formation under hypoxic conditions, providing a negative feedback loop.
What diseases are associated with dysregulated sprouting angiogenesis?
Dysregulation is linked to CNS angiogenesis disorders, abdominal aortic aneurysm, bladder cancer, triple-negative breast cancer, and liver fibrosis.
What methods are used to study negative regulation of sprouting angiogenesis?
Common methods include Transwell migration assays, Matrigel tube formation, endothelial spheroid sprouting, CRISPR knockout/knock-in, and RNA-seq.
Can CRISPR be used to study GO:0090051?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models enable causal interrogation of genes involved in this process.
What is the role of p38 MAPK in angiogenesis?
p38 MAPK is required for mural cell recruitment during angiogenesis, which stabilizes vessels and indirectly limits further sprouting.
How does TRIM15 relate to angiogenesis?
Knockdown of TRIM15 inhibits hepatic stellate cell activation, which is associated with fibrosis and pathological angiogenesis.
Conclusion
GO:0090051, negative regulation of cell migration involved in sprouting angiogenesis, is a critical biological process that restrains endothelial cell migration to ensure proper vessel formation. Key negative regulators such as Nogo-A, EPAC2, DLL4/Notch, and p38 MAPK signaling have been experimentally validated. Dysregulation of this process contributes to cancer, vascular disease, and fibrosis, making it an attractive therapeutic target. CRISPR-based cell models and functional genomics provide powerful tools to dissect the molecular mechanisms underlying this term. By combining knockout, knock-in, point mutation, and overexpression approaches with advanced imaging and bioinformatics, researchers can identify novel negative regulators and translate these findings into clinical applications.
References
- 1. Wälchli T et al.. 2013. Nogo-A is a negative regulator of CNS angiogenesis.. Proc Natl Acad Sci U S A 110(21):E1943-52 PMID: 23625008
- 2. Jing J et al.. 2023. Clinical value of serum miR-1-3p as a potential circulating biomarker for abdominal aortic aneurysm.. Ann Med 55(2):2260395 PMID: 37751480
- 3. Zhang J et al.. 2021. Knockdown of TRIM15 inhibits the activation of hepatic stellate cells.. J Mol Histol 52(4):839-848 PMID: 34142270
- 4. Kim H et al.. 2022. In vitro and in vivo anti-tumor efficacy of krill oil against bladder cancer: Involvement of tumor-associated angiogenic vasculature.. Food Res Int 156:111144 PMID: 35651016
- 5. Ikeda T et al.. 2021. EPAC2 acts as a negative regulator in Matrigel-driven tubulogenesis of human microvascular endothelial cells.. Sci Rep 11(1):19453 PMID: 34593918
- 6. Zhu WH et al.. 2003. Requisite role of p38 MAPK in mural cell recruitment during angiogenesis in the rat aorta model.. J Vasc Res 40(2):140-8 PMID: 12808350
- 7. Liubomirski Y et al.. 2019. Tumor-Stroma-Inflammation Networks Promote Pro-metastatic Chemokines and Aggressiveness Characteristics in Triple-Negative Breast Cancer.. Front Immunol 10:757 PMID: 31031757
- 8. He H et al.. 2011. Effect of DLL4 siRNA on proliferation, migration and tube formation of choroid-retinal endothelial cells under hypoxic conditions.. Chin Med J (Engl) 124(1):118-26 PMID: 21362319
