GO:1905563 negative regulation of vascular endothelial cell proliferation: Signaling Mechanisms, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1905563 describes any biological process that stops, prevents, or reduces the frequency, rate, or extent of vascular endothelial cell proliferation.
• Key signaling pathways include Notch1 activation, KLF15/VASN, ACVR1/ALK2-p21, and G protein subunit alpha i2 (GNAI2).
• Dysregulation of this process contributes to angiogenesis-related diseases such as retinal vasculopathy, cancer, and ischemia.
• CRISPR knockout, point mutation, knock-in, and overexpression models are essential to dissect causal roles of candidate genes in endothelial proliferation.
• High-throughput screening and bioinformatics can identify novel regulators of this GO term for therapeutic targeting.
• Understanding negative regulation is critical for developing anti-angiogenic or pro-angiogenic therapies in cardiovascular and oncological contexts.
Description
The Gene Ontology (GO) term GO:1905563, negative regulation of vascular endothelial cell proliferation, is defined as any process that stops, prevents, or reduces the frequency, rate, or extent of vascular endothelial cell proliferation. Vascular endothelial cells line the inner surface of blood vessels and their controlled proliferation is fundamental to angiogenesis, vascular repair, and homeostasis. This process is tightly regulated by a complex network of signaling pathways, transcription factors, and microRNAs. Dysregulation of negative regulation can lead to pathological angiogenesis, as seen in diabetic retinopathy, tumor growth, and inflammatory disorders. Therefore, understanding the molecular mechanisms that restrain endothelial cell proliferation is of paramount importance for both basic vascular biology and therapeutic development.
negative regulation of vascular endothelial cell proliferation At A Glance
| GO ID | GO:1905563 |
|---|---|
| GO term | negative regulation of vascular endothelial cell proliferation |
| Ontology | biological_process |
| Synonym | down regulation of vascular endothelial cell proliferation, down-regulation of vascular endothelial cell proliferation, downregulation of vascular endothelial cell proliferation, inhibition of vascular endothelial cell proliferation |
| Major function | Stops, prevents, or reduces the frequency, rate, or extent of vascular endothelial cell proliferation |
| Related pathways | Notch signaling, KLF15/VASN axis, ACVR1/ALK2-p21 signaling, G protein subunit alpha i2 (GNAI2) |
| Key regulators | KLF15, VASN, Notch1, ACVR1, p21, GNAI2, miR-24-3p, NKAP, LINC00460, Crip2, RAB3GAP2 |
| Disease relevance | Retinal vasculopathy, cancer, inflammation, ischemia |
What Is GO:1905563?
In simple terms, GO:1905563 encompasses all biological processes that put the brakes on the multiplication of endothelial cells that form blood vessels. According to QuickGO, it includes any mechanism that stops, prevents, or reduces the frequency, rate, or extent of vascular endothelial cell proliferation. This can occur through cell-intrinsic factors (e.g., cell cycle inhibitors, signaling cascades) or extrinsic cues (e.g., secreted molecules, cell-cell interactions).
Why Is negative regulation of vascular endothelial cell proliferation Important in Cell Biology?
Negative regulation of vascular endothelial cell proliferation is crucial for maintaining vascular quiescence and preventing excessive angiogenesis. Imbalances in this process are linked to a wide range of diseases, including cancer, where tumors stimulate uncontrolled endothelial growth, and ischemic disorders, where insufficient angiogenesis impairs tissue repair. Understanding the molecular players and mechanisms provides opportunities for targeted therapies, such as anti-angiogenic drugs or pro-angiogenic factors.
• Prevents pathological angiogenesis in diseases like diabetic retinopathy and cancer.
• Maintains vascular homeostasis and endothelial barrier function.
• Involved in developmental processes such as retinal vascularization.
• Dysregulation contributes to inflammatory diseases and tumor progression.
• Provides targets for anti-angiogenic therapy in oncology.
• Potential to enhance therapeutic angiogenesis in ischemic cardiovascular disease.
• Key to understanding endothelial heterogeneity and organ-specific vascular beds.
• Critical for tissue regeneration and wound healing.
• Serves as a model for studying cell cycle control and signaling crosstalk.
• Enables identification of novel biomarkers and drug targets via CRISPR screens.
What Happens During negative regulation of vascular endothelial cell proliferation?
Initiation by Extracellular Cues
In simple terms: External signals tell endothelial cells to stop dividing.
Negative regulation is often initiated by extracellular ligands such as Notch ligands (e.g., DLL4) or TGF-beta family members. For instance, the KLF15/VASN axis activates Notch1 signaling, which suppresses endothelial proliferation. Similarly, ACVR1/ALK2 signaling via p21 modulates venous endothelial proliferation in the retina.
Receptor Activation and Signal Transduction
In simple terms: Receptors on the cell surface pass the stop signal inside.
Upon ligand binding, receptors like Notch1 are cleaved and translocate to the nucleus to regulate target genes. G protein subunit alpha i2 (GNAI2) also plays a pivotal role in angiogenesis by modulating endothelial cell proliferation through G-protein coupled receptor signaling.
Intracellular Effector Pathways
In simple terms: Inside the cell, specific proteins carry out the stop order.
Key effectors include the cell cycle inhibitor p21, which is induced by ACVR1/ALK2 signaling and blocks cell cycle progression. MicroRNAs such as miR-24-3p can downregulate NKAP and NF-kB signaling to inhibit proliferation under hypoxia. Conversely, LINC00460 promotes proliferation by sponging miR-24-3p, thus relieving inhibition.
Cytoskeletal and Aggregation Changes
In simple terms: Cells may change shape or stick together to halt division.
Crip2 affects vascular development by fine-tuning endothelial cell aggregation and proliferation, suggesting that cytoskeletal dynamics and cell-cell adhesion contribute to negative regulation. RAB3GAP2 regulates skeletal muscle endothelial cell proliferation and capillary-to-fiber ratio, linking vesicular trafficking to proliferative control.
Integration with Angiogenic Sprouting
In simple terms: The stop signal must be coordinated with new blood vessel formation.
Negative regulation is integrated with pro-angiogenic signals such as VEGF. Notch signaling downstream of VEGF induces arterial specification and restricts excessive sprouting, thereby balancing angiogenesis. This integration ensures proper vascular patterning and prevents uncontrolled growth.
Key Genes Involved in GO:1905563 negative regulation of vascular endothelial cell proliferation
The following genes and proteins have been experimentally implicated in the negative regulation of vascular endothelial cell proliferation.
| Gene | Major Role | Research Relevance |
|---|---|---|
| KLF15 | Transcription factor activating VASN and Notch1 signaling | Inhibits angiogenesis; target for anti-angiogenic therapy |
| VASN | Secreted protein activating Notch1 | Mediates KLF15 effects on endothelial proliferation |
| Notch1 | Receptor mediating cell-cell communication | Central to negative regulation and arterial specification |
| ACVR1 | Receptor for BMP signaling | Activates p21 to inhibit venous endothelial proliferation |
| p21 | Cyclin-dependent kinase inhibitor | Effector of ACVR1/ALK2 signaling; blocks cell cycle |
| GNAI2 | G protein subunit alpha i2 | Pivotal role in angiogenesis; modulates proliferation |
| miR-24-3p | MicroRNA | Downregulates NKAP/NF-kB to inhibit proliferation under hypoxia |
| NKAP | NF-kB associated protein | Target of miR-24-3p; promotes proliferation when upregulated |
| LINC00460 | Long non-coding RNA | Sponges miR-24-3p to promote proliferation |
| Crip2 | Cysteine-rich protein 2 | Fine-tunes endothelial aggregation and proliferation |
| RAB3GAP2 | Rab3 GTPase activating protein | Regulates skeletal muscle endothelial proliferation |
| VEGF | Vascular endothelial growth factor | Pro-angiogenic factor; interacts with Notch to balance proliferation |
| DLL4 | Notch ligand | Activates Notch1 to restrict sprouting |
| NF-kB | Transcription factor | Mediates inflammatory and hypoxic responses; modulated by miR-24-3p |
| KLF15/VASN axis | Signaling axis | Inhibits angiogenesis via Notch1 |
| ACVR1/ALK2-p21 axis | Signaling axis | Modulates venous endothelial proliferation |
| GNAI2-GPCR | G protein coupled receptor pathway | Regulates angiogenesis |
| miR-24-3p/NKAP/NF-kB axis | Signaling axis | Responds to IL-1β and hypoxia to inhibit proliferation |
How Is negative regulation of vascular endothelial cell proliferation Regulated?
The negative regulation of vascular endothelial cell proliferation is itself tightly regulated at multiple levels. Transcriptional control includes KLF15 activating VASN and Notch1. Post-transcriptional regulation involves microRNAs such as miR-24-3p, which is modulated by IL-1β and hypoxia and targets NKAP/NF-kB. Long non-coding RNAs like LINC00460 can sponge miR-24-3p, thereby relieving inhibition. Signaling pathways such as ACVR1/ALK2-p21 are responsive to BMP ligands and control cell cycle entry. Additionally, G protein signaling via GNAI2 integrates diverse cues to modulate proliferation. These layers ensure precise control of endothelial quiescence and activation.
negative regulation of vascular endothelial cell proliferation and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ACVR1 | Retinal vasculopathy | Endothelial-specific KO in mouse retina |
| KLF15 | Pathological angiogenesis | Overexpression in zebrafish or mouse |
| GNAI2 | Cancer angiogenesis | Xenograft tumor models with GNAI2 KO |
| miR-24-3p | Inflammatory angiogenesis | Hypoxia-induced endothelial cells with miR-24-3p mimic |
| RAB3GAP2 | Skeletal muscle capillary rarefaction | Muscle-specific KO mouse |
Retinal Vasculopathy
In the retina, ACVR1/ALK2-p21 signaling modulates venous endothelial proliferation, and its dysregulation can lead to pathological retinal angiogenesis, as seen in retinopathy of prematurity and diabetic retinopathy. The KLF15/VASN axis also inhibits angiogenesis, and its loss may contribute to excessive vascular growth.
Cancer
Tumor angiogenesis relies on endothelial cell proliferation. Negative regulators such as Notch1 and GNAI2 can restrict tumor vascularization; their downregulation or mutation may promote tumor growth. Targeting these pathways is a strategy for anti-angiogenic cancer therapy.
Inflammatory and Hypoxic Diseases
IL-1β promotes hypoxic vascular endothelial cell proliferation through the miR-24-3p/NKAP/NF-κB axis, linking inflammation to angiogenesis. LINC00460 stimulates proliferation by downregulating miR-24-3p, suggesting a role in inflammatory angiogenesis.
Skeletal Muscle Capillary Rarefaction
RAB3GAP2 regulates skeletal muscle endothelial cell proliferation and capillary-to-fiber ratio; its dysfunction may contribute to capillary rarefaction in metabolic diseases.
From negative regulation of vascular endothelial cell proliferation-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does gene X inhibit endothelial proliferation? | CRISPR KO in HUVECs followed by proliferation assay |
| Does point mutation in gene Y affect its function? | Knock-in of point mutant in endothelial cells |
| Does overexpression of gene Z suppress angiogenesis? | Lentiviral overexpression in mouse retina |
| What is the role of gene W in developmental angiogenesis? | Zebrafish knockout or knock-in |
| Can we identify novel negative regulators? | Genome-wide CRISPR library screening in endothelial cells |
| Does gene V regulate capillary density in muscle? | Endothelial-specific inducible KO mouse |
How to Study the negative regulation of vascular endothelial cell proliferation Process
| Method | What It Measures | Typical Application |
|---|---|---|
| CRISPR KO screening | Gene essentiality for proliferation | Identify novel negative regulators |
| RNA-seq | Transcriptional changes | Downstream effects of KLF15 overexpression |
| Phosphoproteomics | Signaling pathway activity | ACVR1/ALK2-p21 axis activation |
| EdU incorporation | DNA synthesis rate | Quantify endothelial proliferation |
| Retinal whole-mount | Vascular density and sprouting | In vivo angiogenesis models |
| Spheroid sprouting | Endothelial sprouting capacity | Assess negative regulation in 3D |
| Luciferase reporter | miRNA target regulation | miR-24-3p/NKAP interaction |
| Chromatin immunoprecipitation | Transcription factor binding | Notch1 target gene regulation |
CRISPR Screening
Genome-wide CRISPR knockout or activation screens in primary endothelial cells can identify genes whose loss or gain alters proliferation under baseline or stimulated conditions. This unbiased approach has uncovered microRNA and lncRNA regulators.
Transcriptomics and Epigenomics
RNA-seq and ATAC-seq of endothelial cells with perturbations in candidate genes (e.g., KLF15, ACVR1) reveal downstream transcriptional programs and chromatin accessibility changes that mediate negative regulation.
Proteomics and Phosphoproteomics
Mass spectrometry-based proteomics can quantify changes in signaling pathways (e.g., Notch, BMP) and identify post-translational modifications on key effectors like p21 after induction of negative regulation.
Imaging and Functional Assays
Live-cell imaging of proliferation markers (e.g., Ki-67, EdU) combined with spheroid sprouting assays or retinal whole-mount staining allows spatial and temporal assessment of negative regulation in vitro and in vivo.
How CRISPR Can Be Used to Study GO:1905563 negative regulation of vascular endothelial cell proliferation
Knockout
CRISPR knockout of candidate negative regulators (e.g., KLF15, ACVR1) in endothelial cells or mouse models can confirm their role in suppressing proliferation. For example, ACVR1 knockout in retinal endothelial cells increases proliferation.
Point Mutation
Introducing point mutations (e.g., in the Notch1 cleavage site or p21 phosphorylation sites) via CRISPR knock-in allows precise dissection of signaling events without complete loss of protein.
Knock-in
Knock-in of tagged versions (e.g., GFP, HA) of genes like VASN or GNAI2 enables live-cell imaging and biochemical isolation of protein complexes to study their function in negative regulation.
Overexpression
CRISPR activation (CRISPRa) or lentiviral overexpression of negative regulators such as KLF15 or miR-24-3p can suppress endothelial proliferation and angiogenesis in vitro and in vivo.
How EDITGENE Supports negative regulation of vascular endothelial cell proliferation Research
Researchers studying negative regulation of vascular endothelial cell proliferation-related genes often need to determine whether a candidate gene is causally involved in suppressing endothelial growth. This requires precise genetic manipulation and functional validation. EDITGENE provides a comprehensive suite of CRISPR services to accelerate such investigations.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of vascular endothelial cell proliferation research.
Frequently Asked Questions About negative regulation of vascular endothelial cell proliferation
What is GO:1905563?
GO:1905563 is the Gene Ontology term for negative regulation of vascular endothelial cell proliferation, defined as any process that stops, prevents, or reduces the frequency, rate, or extent of vascular endothelial cell proliferation.
What genes are involved in negative regulation of vascular endothelial cell proliferation?
Key genes include KLF15, VASN, Notch1, ACVR1, p21, GNAI2, miR-24-3p, NKAP, LINC00460, Crip2, and RAB3GAP2.
How does Notch signaling inhibit endothelial cell proliferation?
Notch1 activation by ligands such as DLL4 leads to cleavage and nuclear translocation of the Notch intracellular domain, which regulates target genes that suppress proliferation and promote arterial specification.
What is the role of ACVR1/ALK2-p21 signaling in endothelial cells?
ACVR1/ALK2 signaling induces p21, a cyclin-dependent kinase inhibitor, which blocks cell cycle progression and inhibits venous endothelial proliferation in the retinal vasculature.
How do microRNAs regulate endothelial cell proliferation?
MicroRNAs such as miR-24-3p can downregulate NKAP and NF-kB signaling to inhibit proliferation under hypoxia, while lncRNAs like LINC00460 can sponge miR-24-3p to promote proliferation.
What diseases are associated with dysregulation of negative regulation of vascular endothelial cell proliferation?
Diseases include retinal vasculopathy, cancer, inflammatory angiogenesis, and skeletal muscle capillary rarefaction.
What research methods are used to study negative regulation of vascular endothelial cell proliferation?
Methods include CRISPR screening, RNA-seq, proteomics, imaging, and functional assays such as EdU incorporation and spheroid sprouting.
How can CRISPR be used to study negative regulation of vascular endothelial cell proliferation?
CRISPR knockout, point mutation, knock-in, and overexpression models allow precise manipulation of candidate genes to test their causal role in suppressing endothelial proliferation.
What is the role of GNAI2 in angiogenesis?
G protein subunit alpha i2 (GNAI2) plays a pivotal role in angiogenesis by modulating endothelial cell proliferation through G-protein coupled receptor signaling.
How does KLF15 inhibit angiogenesis?
KLF15 activates the expression of VASN, which in turn activates Notch1 signaling, leading to inhibition of endothelial cell proliferation and angiogenesis.
Conclusion
Negative regulation of vascular endothelial cell proliferation (GO:1905563) is a critical biological process that maintains vascular homeostasis and prevents pathological angiogenesis. Key signaling pathways such as Notch1, ACVR1/ALK2-p21, and GNAI2, along with microRNAs and lncRNAs, orchestrate this regulation. Dysregulation contributes to diseases including retinal vasculopathy, cancer, and inflammatory disorders. Advances in CRISPR-based models and high-throughput screening are accelerating the discovery of novel regulators and therapeutic targets. Continued research into this process holds promise for developing precision therapies for angiogenesis-related diseases.
References
- 1. Zhang J et al.. 2025. Endothelial KLF15/VASN Axis Inhibits Angiogenesis via Activation of Notch1 Signaling.. Circ Res 136(12):1595-1609 PMID: 40297901
- 2. Yang S et al.. 2025. Crip2 affects vascular development by fine-tuning endothelial cell aggregation and proliferation.. Cell Mol Life Sci 82(1):110 PMID: 40074973
- 3. Huang J et al.. 2022. IL-1β promotes hypoxic vascular endothelial cell proliferation through the miR-24-3p/NKAP/NF-κB axis.. Biosci Rep 42(1) PMID: 35005769
- 4. Jia R et al.. 2022. LINC00460 Stimulates the Proliferation of Vascular Endothelial Cells by Downregulating miRNA-24-3p.. Dis Markers 2022:2524156 PMID: 35222741
- 5. Ström K et al.. 2026. RAB3GAP2 is a regulator of skeletal muscle endothelial cell proliferation and associated with capillary-to-fiber ratio.. Cell Rep 45(2):116961 PMID: 41678332
- 6. Bai CW et al.. 2024. G protein subunit alpha i2's pivotal role in angiogenesis.. Theranostics 14(5):2190-2209 PMID: 38505600
- 7. Hirashima M. 2009. Regulation of endothelial cell differentiation and arterial specification by VEGF and Notch signaling.. Anat Sci Int 84(3):95-101 PMID: 19259767
- 8. Pak B et al.. 2024. ACVR1/ALK2-p21 signaling axis modulates proliferation of the venous endothelium in the retinal vasculature.. Angiogenesis 27(4):765-777 PMID: 38955953
