GO:1903141 negative regulation of establishment of endothelial barrier: Barrier Breakdown Mechanisms, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1903141 describes any process that stops, prevents, or reduces the establishment of the endothelial barrier, a critical structure controlling vascular permeability.
• Loss of endothelial barrier integrity is a hallmark of blood-brain barrier damage in aging, diabetic intracerebral hemorrhage, and neuroinflammation.
• Key molecular players include CX43, PARP1, FUNDC1, ISG15, PKC-β, and focal adhesion kinase (FAK), which modulate barrier function through distinct signaling axes.
• Shear stress and the innate immune system are potent regulators of endothelial barrier establishment and its negative regulation.
• CRISPR-based models (knockout, point mutation, knock-in, overexpression) enable causal dissection of genes that negatively regulate endothelial barrier establishment.
• Understanding GO:1903141 has therapeutic implications for stroke, diabetic hemorrhage, neurodegeneration, and cancer immunotherapy.
Description
The endothelial barrier is a highly specialized structure that lines blood vessels and controls the passage of molecules and cells between the bloodstream and surrounding tissues. Its establishment requires coordinated signaling events that strengthen cell-cell junctions and focal adhesions. GO:1903141, negative regulation of establishment of endothelial barrier, captures the biological processes that oppose or reduce this barrier-forming program. This term is essential for understanding how pathological conditions such as aging, diabetes, and inflammation disrupt vascular integrity. Recent studies have identified specific molecular axes, including the CX43-PARP1 pathway in aging-induced blood-brain barrier damage and the Isg15-FUNDC1 axis in diabetic intracerebral hemorrhage, that actively suppress barrier establishment. Protein kinase C-β and focal adhesion kinase have also been shown to differentially regulate barrier-forming capacity in brain microvascular endothelial cells. Researchers studying vascular biology, neuroinflammation, and cancer immunotherapy need to understand GO:1903141 to identify therapeutic targets that preserve or restore endothelial barrier function.
negative regulation of establishment of endothelial barrier At A Glance
| GO ID | GO:1903141 |
|---|---|
| GO term | negative regulation of establishment of endothelial barrier |
| Ontology | biological_process |
| Synonym | down regulation of establishment of endothelial barrier, down-regulation of establishment of endothelial barrier, downregulation of establishment of endothelial barrier, inhibition of establishment of endothelial barrier |
| Major function | Inhibition or reduction of endothelial barrier formation, leading to increased vascular permeability |
| Related processes | Blood-brain barrier damage, vascular permeability, inflammation, aging, diabetic hemorrhage |
| Key regulators | CX43, PARP1, FUNDC1, ISG15, PKC-β, focal adhesion kinase (FAK) |
| Disease relevance | Stroke, diabetic intracerebral hemorrhage, neurodegeneration, cancer immunotherapy |
What Is GO:1903141?
GO:1903141 is a biological process term defined as any process that stops, prevents, or reduces the frequency, rate, or extent of establishment of endothelial barrier. In other words, it encompasses molecular and cellular events that actively inhibit the formation or maintenance of the endothelial barrier, leading to increased vascular permeability. This term is distinct from positive regulation of endothelial barrier establishment and is often studied in the context of blood-brain barrier breakdown, inflammation, and disease.
Why Is negative regulation of establishment of endothelial barrier Important in Cell Biology?
GO:1903141 is critically important because endothelial barrier dysfunction underlies a wide range of human diseases, including ischemic stroke, diabetic hemorrhage, neurodegeneration, and cancer metastasis. Understanding the negative regulatory mechanisms that suppress barrier establishment can reveal therapeutic targets to stabilize vascular integrity. For example, NAD+ rescue of aging-induced blood-brain barrier damage via the CX43-PARP1 axis highlights a druggable pathway. Similarly, semaglutide targeting the Isg15-FUNDC1 axis attenuates blood-brain barrier injury in diabetic intracerebral hemorrhage. These findings underscore the clinical relevance of this GO term.
• Endothelial barrier breakdown is a hallmark of blood-brain barrier damage in aging and neurodegenerative diseases.
• Diabetic intracerebral hemorrhage involves IFN-β overactivation and FUNDC1-mediated barrier injury.
• Protein kinase C-β differentially regulates barrier-forming capacity in brain microvascular endothelial cells.
• Focal adhesion kinase activation strengthens endothelial barrier, and its negative regulation contributes to permeability.
• Shear stress is a critical physiological regulator of blood-brain barrier endothelial physiology.
• Innate immune barriers influence antitumor immunity and endothelial barrier function.
• Endothelial cells play key roles in mammary gland health and disease, including barrier regulation.
• Targeting negative regulators of endothelial barrier establishment may treat stroke, edema, and inflammation.
• CRISPR screening can identify novel genes that negatively regulate endothelial barrier establishment.
• Understanding GO:1903141 aids in developing therapies for vascular leak syndromes and cancer immunotherapy.
What Happens During negative regulation of establishment of endothelial barrier?
Initiation of Barrier Destabilization
In simple terms: The barrier starts to weaken when specific signals tell endothelial cells to loosen their connections.
Negative regulation of endothelial barrier establishment can be initiated by pathological stimuli such as aging, hyperglycemia, or inflammatory cytokines. In aging, decreased NAD+ levels lead to PARP1 activation and CX43 dysfunction, which disrupts blood-brain barrier integrity. In diabetic intracerebral hemorrhage, IFN-β overactivation via the Isg15-FUNDC1 axis promotes barrier injury. These initial signals set off cascades that reduce the frequency or extent of barrier formation.
Signaling Cascades That Suppress Barrier Formation
In simple terms: A series of molecular signals actively block the proteins that would normally build a strong barrier.
Protein kinase C-β (PKC-β) distinctly regulates the blood-brain barrier-forming capacity of brain microvascular endothelial cells and outgrowth endothelial cells. Focal adhesion kinase (FAK) activation is normally required for endothelial barrier strengthening, so its inhibition or negative regulation contributes to barrier suppression. The CX43-PARP1 axis and Isg15-FUNDC1 axis represent specific signaling cascades that negatively regulate barrier establishment.
Cytoskeletal and Junctional Rearrangements
In simple terms: The cell's internal skeleton and the junctions between cells get reorganized to open up gaps.
Negative regulation of endothelial barrier establishment often involves reorganization of the actin cytoskeleton and disruption of adherens and tight junctions. FAK, a key regulator of focal adhesions, when negatively regulated, leads to weakened cell-matrix attachments and increased permeability. Shear stress can modulate these rearrangements, influencing blood-brain barrier endothelial physiology.
Immune and Inflammatory Amplification
In simple terms: Immune signals can make the barrier leaky, and this process can feed back to cause more damage.
Innate immune barriers, including those involving IFN-β, can amplify endothelial barrier disruption. In diabetic intracerebral hemorrhage, IFN-β overactivation via the Isg15-FUNDC1 axis exacerbates blood-brain barrier injury. Lifting innate immune barriers is also relevant to antitumor immunity, where endothelial barrier function affects immune cell infiltration.
Resolution or Chronic Dysfunction
In simple terms: Depending on the context, the barrier may recover, or the damage may become long-lasting.
The outcome of negative regulation of endothelial barrier establishment depends on the persistence of the initiating stimulus. In aging, NAD+ supplementation can rescue blood-brain barrier damage via the CX43-PARP1 axis, indicating reversibility. In diabetic intracerebral hemorrhage, semaglutide targeting the Isg15-FUNDC1 axis attenuates barrier injury, suggesting a therapeutic window. Chronic dysfunction contributes to neurodegeneration and edema.
Key Genes Involved in GO:1903141 negative regulation of establishment of endothelial barrier
The following genes and proteins have been experimentally implicated in the negative regulation of endothelial barrier establishment, based on verified PubMed literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| CX43 | Gap junction protein; NAD+ rescue of aging-induced BBB damage via CX43-PARP1 axis | Target for aging-related BBB dysfunction |
| PARP1 | DNA repair enzyme; activated in aging, contributes to BBB damage | Therapeutic target for NAD+ rescue |
| ISG15 | Interferon-stimulated gene; part of Isg15-FUNDC1 axis in diabetic ICH | Target for semaglutide in diabetic hemorrhage |
| FUNDC1 | Mitochondrial protein; mediates IFN-β overactivation and BBB injury | Target for attenuating BBB injury |
| PKC-β | Protein kinase C beta; distinctly regulates BBB-forming capacity | Modulator of endothelial barrier in BMECs |
| FAK | Focal adhesion kinase; activation strengthens endothelial barrier | Target for barrier strengthening |
| IFN-β | Inflammatory cytokine; overactivation in diabetic ICH | Biomarker and therapeutic target |
| NAD+ | Cofactor; rescues aging-induced BBB damage | Nutraceutical intervention |
| Semaglutide | GLP-1 receptor agonist; targets Isg15-FUNDC1 axis | Drug repurposing for diabetic ICH |
| Shear stress | Mechanical force; regulates BBB endothelial physiology | Biophysical modulator |
| Endothelial cells | Cellular building blocks of the barrier | Primary cell models |
| Innate immune system | Barriers to antitumor immunity | Cancer immunotherapy target |
| Adherens junctions | Cell-cell adhesion structures | Structural targets |
| Tight junctions | Barrier-forming junctions | Structural targets |
| Actin cytoskeleton | Cytoskeletal network | Dynamic regulator |
| VEGF | Vascular endothelial growth factor; increases permeability | Angiogenesis and permeability |
| TNF-α | Inflammatory cytokine; disrupts barrier | Inflammation model |
How Is negative regulation of establishment of endothelial barrier Regulated?
The negative regulation of endothelial barrier establishment is controlled by multiple signaling pathways. The CX43-PARP1 axis is regulated by NAD+ levels, which decline with aging. The Isg15-FUNDC1 axis is modulated by IFN-β and is a target of semaglutide. Protein kinase C-β differentially regulates barrier-forming capacity in brain microvascular endothelial cells. Focal adhesion kinase activity is required for barrier strengthening, and its negative regulation leads to increased permeability. Shear stress also acts as a physiological regulator of blood-brain barrier endothelial physiology. Innate immune signaling can lift barriers to antitumor immunity, indirectly affecting endothelial barrier function.
negative regulation of establishment of endothelial barrier and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CX43 | Aging-induced BBB damage | Knockout mouse, NAD+ supplementation |
| PARP1 | Aging-induced BBB damage | PARP1 inhibitor, knockout |
| ISG15 | Diabetic intracerebral hemorrhage | Knockout mouse, semaglutide treatment |
| FUNDC1 | Diabetic intracerebral hemorrhage | Knockout mouse, IFN-β modulation |
| PKC-β | BBB dysfunction | BMEC and OEC models, PKC-β inhibitors |
Aging-Induced Blood-Brain Barrier Damage
Aging is associated with decreased NAD+ levels, which leads to blood-brain barrier damage via the CX43-PARP1 axis. This negative regulation of endothelial barrier establishment contributes to neurodegeneration and cognitive decline. NAD+ supplementation rescues this damage, highlighting a potential therapeutic strategy.
Diabetic Intracerebral Hemorrhage
In diabetic intracerebral hemorrhage, IFN-β overactivation via the Isg15-FUNDC1 axis promotes blood-brain barrier injury. Semaglutide targets this axis, suppressing IFN-β overactivation and attenuating barrier damage. This demonstrates a direct link between metabolic disease and negative regulation of endothelial barrier establishment.
Cancer and Immunotherapy
Endothelial barrier function affects immune cell infiltration into tumors. Lifting innate immune barriers to antitumor immunity involves modulating endothelial barrier establishment. Negative regulation of this barrier can either promote or inhibit tumor progression depending on context, making it a complex therapeutic target.
Skin Vascular Permeability
Regulation of blood vascular permeability in the skin is critical for inflammatory responses and tissue homeostasis. Negative regulation of endothelial barrier establishment in skin microvessels contributes to edema and inflammation. Understanding these mechanisms may lead to treatments for skin inflammatory diseases.
From negative regulation of establishment of endothelial barrier-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does gene X negatively regulate endothelial barrier establishment? | CRISPR knockout in brain microvascular endothelial cells |
| Does a specific point mutation in gene Y affect barrier function? | CRISPR point mutation knock-in |
| Can a tagged version of protein Z reveal its localization during barrier disruption? | CRISPR knock-in with fluorescent tag |
| Does overexpression of gene W suppress barrier establishment? | CRISPR overexpression (CRISPRa) or lentiviral overexpression |
| Which genes are essential for barrier establishment? | Genome-wide CRISPR library screening |
| What are the transcriptomic changes during barrier disruption? | RNA-seq after CRISPR knockout |
How to Study the negative regulation of establishment of endothelial barrier Process
| Method | What It Measures | Typical Application |
|---|---|---|
| RNA-seq | Global gene expression changes | Identify pathways in barrier disruption |
| Proteomics | Protein abundance and modifications | Quantify PARP1, FAK, ISG15 |
| TEER assay | Barrier integrity | Functional validation of CRISPR knockouts |
| Permeability assay | Paracellular flux | Assess barrier leakiness |
| Immunofluorescence | Protein localization | Visualize junctional disruption |
| CRISPR library screening | Gene essentiality | Discover novel negative regulators |
| Phosphoproteomics | Signaling events | Map kinase cascades |
Transcriptomic Profiling
RNA sequencing (RNA-seq) can identify gene expression changes during negative regulation of endothelial barrier establishment. For example, comparing endothelial cells under aging or diabetic conditions to controls reveals pathways such as CX43-PARP1 or Isg15-FUNDC1. This method helps pinpoint candidate genes for further CRISPR validation.
Proteomic and Phosphoproteomic Analysis
Mass spectrometry-based proteomics can quantify protein abundance and post-translational modifications, such as PARP1 activation or FAK phosphorylation, during barrier disruption. Phosphoproteomics specifically identifies signaling events that negatively regulate barrier establishment.
Functional Barrier Assays
Transendothelial electrical resistance (TEER) and permeability assays using fluorescent tracers measure barrier integrity in vitro. These assays are essential to confirm that a gene or treatment negatively regulates barrier establishment. They can be combined with CRISPR knockout or overexpression.
Imaging and Localization Studies
Immunofluorescence and live-cell imaging can visualize junctional proteins, cytoskeletal rearrangements, and protein localization during barrier disruption. For example, tracking CX43 or FUNDC1 localization provides mechanistic insights.
How CRISPR Can Be Used to Study GO:1903141 negative regulation of establishment of endothelial barrier
Knockout
CRISPR knockout of candidate genes such as CX43, PARP1, ISG15, or FUNDC1 in endothelial cells can determine whether they are required for negative regulation of barrier establishment. For example, PARP1 knockout may rescue aging-induced barrier damage. Knockout models are essential for causal inference.
Point Mutation
CRISPR point mutation can introduce specific amino acid changes to test the function of domains or phosphorylation sites in proteins like FAK or PKC-β. This approach reveals mechanistic details of how these proteins regulate barrier establishment.
Knock-in
CRISPR knock-in of fluorescent tags or reporter genes allows real-time tracking of proteins such as CX43 or FUNDC1 during barrier disruption. Tagged knock-in models are valuable for imaging and biochemical studies.
Overexpression
CRISPR activation (CRISPRa) or lentiviral overexpression can test whether increasing the levels of a candidate gene suppresses barrier establishment. For example, overexpressing ISG15 or FUNDC1 may exacerbate barrier injury. Overexpression models complement knockout studies.
How EDITGENE Supports negative regulation of establishment of endothelial barrier Research
Researchers studying negative regulation of establishment of endothelial barrier-related genes often need to determine whether a candidate gene is causally involved in barrier disruption or protection. EDITGENE provides comprehensive CRISPR services to accelerate this discovery process.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of establishment of endothelial barrier research.
Frequently Asked Questions About negative regulation of establishment of endothelial barrier
What is GO:1903141?
GO:1903141 is a Gene Ontology biological process term defined as any process that stops, prevents, or reduces the frequency, rate, or extent of establishment of endothelial barrier.
What genes are involved in negative regulation of endothelial barrier establishment?
Key genes include CX43, PARP1, ISG15, FUNDC1, PKC-β, and FAK, as identified in recent studies.
How is the endothelial barrier negatively regulated in aging?
Aging reduces NAD+ levels, leading to PARP1 activation and CX43 dysfunction, which disrupts the blood-brain barrier.
What is the role of the Isg15-FUNDC1 axis in diabetic intracerebral hemorrhage?
The Isg15-FUNDC1 axis mediates IFN-β overactivation and blood-brain barrier injury in diabetic intracerebral hemorrhage.
Can semaglutide protect the blood-brain barrier?
Semaglutide targets the Isg15-FUNDC1 axis, suppressing IFN-β overactivation and attenuating blood-brain barrier injury in diabetic intracerebral hemorrhage.
How does protein kinase C-β regulate endothelial barrier?
PKC-β distinctly regulates the blood-brain barrier-forming capacity of brain microvascular endothelial cells and outgrowth endothelial cells.
What is the role of focal adhesion kinase in endothelial barrier?
FAK activation strengthens the endothelial barrier, and its negative regulation contributes to increased permeability.
How does shear stress affect the blood-brain barrier?
Shear stress is a critical regulator of blood-brain barrier endothelial physiology, influencing barrier establishment.
What diseases are associated with negative regulation of endothelial barrier establishment?
Diseases include aging-induced blood-brain barrier damage, diabetic intracerebral hemorrhage, neurodegeneration, and cancer.
How can CRISPR be used to study GO:1903141?
CRISPR knockout, point mutation, knock-in, and overexpression models allow causal testing of genes involved in negative regulation of endothelial barrier establishment.
Conclusion
GO:1903141, negative regulation of establishment of endothelial barrier, is a critical biological process that underlies vascular dysfunction in aging, diabetes, and cancer. Recent studies have identified specific molecular axes, such as CX43-PARP1 and Isg15-FUNDC1, that actively suppress barrier formation. Understanding these mechanisms offers therapeutic opportunities to stabilize the endothelial barrier and treat related diseases. CRISPR-based models and EDITGENE services provide powerful tools to dissect these pathways and accelerate discovery.
References
- 1. Zhan R et al.. 2023. NAD(+) rescues aging-induced blood-brain barrier damage via the CX43-PARP1 axis.. Neuron 111(22):3634-3649.e7 PMID: 37683629
- 2. Wang Y et al.. 2026. Semaglutide targets the Isg15-FUNDC1 axis: suppressing IFN-β overactivation and attenuating blood-brain barrier injury in diabetic intracerebral hemorrhage.. J Neuroinflammation 23(1) PMID: 42186030
- 3. Kadir RRA et al.. 2022. Protein kinase C-β distinctly regulates blood-brain barrier-forming capacity of Brain Microvascular endothelial cells and outgrowth endothelial cells.. Metab Brain Dis 37(6):1815-1827 PMID: 35763197
- 4. Ono S et al.. 2017. Regulation of blood vascular permeability in the skin.. Inflamm Regen 37:11 PMID: 29259710
- 5. Quadri SK et al.. 2003. Endothelial barrier strengthening by activation of focal adhesion kinase.. J Biol Chem 278(15):13342-9 PMID: 12556538
- 6. Cucullo L et al.. 2011. The role of shear stress in Blood-Brain Barrier endothelial physiology.. BMC Neurosci 12:40 PMID: 21569296
- 7. Ryman VE et al.. 2015. Role of endothelial cells in bovine mammary gland health and disease.. Anim Health Res Rev 16(2):135-49 PMID: 26303748
- 8. Rothlin CV et al.. 2020. Lifting the innate immune barriers to antitumor immunity.. J Immunother Cancer 8(1) PMID: 32273348
