GO:0120331 endothelial tube formation: Developmental Process, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0120331 endothelial tube formation describes the developmental process pertaining to the initial formation of an endothelial tube.
• Endothelial tube formation is a fundamental step in vasculogenesis and angiogenesis, requiring coordinated endothelial cell migration, adhesion, and morphogenesis [2, 4].
• Key molecular players include cadherin 5 (CDH5), CD31 (PECAM1), connexin32 (GJB1), and signaling through Hedgehog, PI3K, and MAPK/ERK pathways [1, 4, 5, 6, 8].
• Dysregulation of endothelial tube formation contributes to diabetic vascular complications, tumor angiogenesis, and lymphatic disorders [3, 7, 8].
• CRISPR-based knockout, knock-in, and overexpression models enable causal interrogation of genes involved in endothelial tube formation.
• EDITGENE provides comprehensive CRISPR services including library screening and bioinformatics to accelerate vascular biology research.
Description
Endothelial tube formation is the developmental process pertaining to the initial formation of an endothelial tube, a critical event in the establishment of the vascular system. This process is essential for vasculogenesis, the de novo formation of blood vessels, and angiogenesis, the sprouting of new vessels from existing ones [2, 4]. Understanding the molecular and cellular mechanisms governing endothelial tube formation is fundamental to vascular biology and has broad implications for human health and disease [2, 3, 8]. Researchers study endothelial tube formation to uncover how endothelial cells coordinate their behavior to form hollow tubes, a process that requires precise regulation of cell adhesion, migration, and signaling [2, 5, 6]. Defects in this process are linked to a range of pathological conditions, including diabetic vascular damage, tumor progression, and lymphatic dysfunction [3, 7, 8]. The QuickGO definition (GO:0120331) provides a precise ontological framework for annotating genes and proteins involved in this developmental process. This article synthesizes authoritative QuickGO data and verified PubMed literature to deliver a research-grade overview of endothelial tube formation, covering its definition, molecular mechanisms, key genes, disease relevance, and experimental models. It is designed for researchers seeking to understand or manipulate this process using CRISPR-based approaches.
endothelial tube formation At A Glance
| GO ID | GO:0120331 |
|---|---|
| GO term | endothelial tube formation |
| Ontology | biological_process |
| Synonym | None |
| Definition | The developmental process pertaining to the initial formation of an endothelial tube. |
| Major function | Formation of the primary endothelial tube during vasculogenesis and angiogenesis. |
| Related processes | Vasculogenesis, angiogenesis, endothelial cell migration, cell adhesion. |
| Key cellular events | Endothelial cell migration, adhesion, lumen formation, tube morphogenesis. |
What Is GO:0120331?
GO:0120331 endothelial tube formation is defined by QuickGO as the developmental process pertaining to the initial formation of an endothelial tube. In simpler terms, it encompasses the cellular and molecular events that lead to the creation of a hollow, tube-like structure composed of endothelial cells, which is the foundational step in building blood and lymphatic vessels. This process is distinct from later stages of vessel maturation and remodeling, focusing specifically on the initial morphogenetic events that generate the endothelial tube [2, 4].
Why Is endothelial tube formation Important in Cell Biology?
Endothelial tube formation is a cornerstone of vascular development and homeostasis, and its dysregulation is implicated in numerous human diseases [2, 3, 8]. Understanding this process at the molecular level is essential for developing therapeutic strategies targeting angiogenesis in cancer, diabetic vascular complications, and lymphatic disorders [3, 7, 8]. Moreover, the ability to manipulate genes involved in endothelial tube formation using CRISPR technologies offers unprecedented opportunities to dissect causal mechanisms and identify novel drug targets [1, 4, 5, 6].
• Critical for embryonic development and organogenesis, as endothelial tubes form the primitive vascular network [2, 4].
• Central to angiogenesis, which supports tumor growth and metastasis; targeting tube formation is a therapeutic strategy in oncology [7, 8].
• Dysregulated in diabetic vascular complications, including endothelial damage and impaired angiogenesis.
• Involved in lymphatic vessel formation; inhibition of lymphatic endothelial tube formation is relevant to lymphedema and tumor lymphangiogenesis.
• Requires coordinated signaling through Hedgehog, PI3K, MAPK/ERK, and VEGFR pathways [4, 6, 8].
• Cell adhesion molecules such as cadherin 5 and CD31 are essential for tube formation, linking cytoskeletal dynamics to morphogenesis.
• Connexin32 enhances endothelial tube formation and migration, highlighting the role of gap junctions.
• Decorin inhibits endothelial migration and tube-like structure formation via thrombospondin-1, indicating extracellular matrix regulation.
• Stromal cell-derived factor-1alpha induces tube-like structure formation through PI3K, linking chemokines to vascular morphogenesis.
• CRISPR screening and knockout models enable systematic discovery of genes regulating endothelial tube formation.
What Happens During endothelial tube formation?
Initiation and endothelial cell activation
In simple terms: Endothelial cells receive signals to start forming a tube.
Endothelial tube formation begins with the activation of endothelial cells by pro-angiogenic factors, leading to changes in gene expression and cell behavior [2, 4]. Hedgehog signaling is essential for endothelial tube formation during vasculogenesis, as demonstrated in mouse embryos where disruption of Hedgehog signaling prevents tube formation. This initiation phase involves the upregulation of adhesion molecules and the secretion of matrix metalloproteinases to remodel the extracellular matrix.
Cell migration and alignment
In simple terms: Endothelial cells move and line up to form the initial cord.
Following activation, endothelial cells migrate and align to form cord-like structures. Stromal cell-derived factor-1alpha induces tube-like structure formation through phosphoinositide 3-kinase (PI3K) signaling, highlighting the role of chemokine gradients in directing migration. Connexin32 enhances endothelial tube formation and cell migration, suggesting that gap junction communication coordinates collective cell movement. Decorin inhibits endothelial migration and tube-like structure formation via thrombospondin-1, indicating that extracellular matrix components can negatively regulate this step.
Adhesion and lumen formation
In simple terms: Cells stick together and create a hollow space inside the tube.
Cell-cell adhesion is critical for stabilizing the nascent tube and forming a lumen. Endothelial cell tube formation depends on cadherin 5 and CD31 interactions with filamentous actin, which mediate cytoskeletal rearrangements necessary for morphogenesis. These adhesion complexes coordinate the polarization of endothelial cells and the creation of a central lumen. The process is further regulated by signaling pathways such as MAPK/ERK, which can be modulated by paracrine factors from mesenchymal stem cells to protect endothelial function.
Tube stabilization and maturation
In simple terms: The tube becomes stable and ready to carry blood or lymph.
Once the initial tube is formed, it undergoes stabilization and maturation, involving the recruitment of mural cells and deposition of basement membrane. VEGFR-3-mediated ERK signaling is required for lymphatic endothelial tube formation, and its inhibition by ciclopirox blocks this process. Proper stabilization ensures the tube can withstand hemodynamic forces and function as a conduit.
Key Genes Involved in GO:0120331 endothelial tube formation
The following genes and proteins have been experimentally implicated in endothelial tube formation, as supported by the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| GJB1 (Connexin32) | Enhances endothelial tube formation and cell migration | Gap junction communication in angiogenesis |
| CDH5 (Cadherin 5) | Mediates endothelial cell-cell adhesion and lumen formation | Essential for tube formation via actin cytoskeleton |
| PECAM1 (CD31) | Endothelial cell adhesion molecule interacting with cadherin 5 | Required for tube formation and actin dynamics |
| SHH (Sonic Hedgehog) | Signaling molecule essential for endothelial tube formation during vasculogenesis | Embryonic vascular development |
| CXCL12 (SDF-1alpha) | Chemokine inducing tube-like structure formation via PI3K | Chemokine-driven angiogenesis |
| PIK3CA (PI3K) | Kinase mediating SDF-1alpha-induced tube formation | Intracellular signaling in endothelial morphogenesis |
| DCN (Decorin) | Inhibits endothelial migration and tube-like structure formation | Extracellular matrix regulation of angiogenesis |
| THBS1 (Thrombospondin-1) | Mediates decorin-induced inhibition of tube formation | Anti-angiogenic factor |
| VEGFR3 (FLT4) | Receptor for lymphatic endothelial tube formation | Lymphangiogenesis and ERK signaling |
| MAPK1/3 (ERK) | Kinase pathway downstream of VEGFR-3 and other receptors | Regulates tube formation in lymphatic and blood endothelium [3, 8] |
| MAP2K1 (MEK) | Upstream kinase in MAPK/ERK pathway | Paracrine protection of endothelium |
| KDR (VEGFR2) | Major pro-angiogenic receptor | Angiogenesis signaling (implied by [2, 4]) |
| FLT1 (VEGFR1) | Modulates VEGF signaling | Angiogenesis regulation (implied by [2, 4]) |
| NOTCH1 | Regulates endothelial cell fate and sprouting | Vascular development (implied by) |
| DLL4 | Notch ligand involved in tip/stalk cell selection | Angiogenesis (implied by) |
| ANGPT2 (Angiopoietin-2) | Regulates vessel stability and sprouting | Vascular remodeling (implied by) |
| TEK (TIE2) | Endothelial receptor tyrosine kinase | Vessel stabilization (implied by) |
How Is endothelial tube formation Regulated?
Endothelial tube formation is regulated by a complex interplay of signaling pathways. Hedgehog signaling is essential during vasculogenesis. The PI3K pathway mediates chemokine-induced tube formation, while the MAPK/ERK pathway is involved in both blood and lymphatic endothelial tube formation and can be modulated by paracrine factors [3, 8]. Additionally, gap junction proteins such as connexin32 enhance tube formation, and extracellular matrix components like decorin and thrombospondin-1 can inhibit it. These regulatory mechanisms ensure tight spatial and temporal control of vascular morphogenesis.
endothelial tube formation and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| MAPK1/3 | Diabetic vascular damage | Knockout or knock-in of MAPK1/3 in endothelial cells to study paracrine protection |
| VEGFR3 (FLT4) | Lymphatic disorders and tumor lymphangiogenesis | Knockout or point mutation to block ERK signaling and tube formation |
| CDH5 | Vascular malformations | Knockout or point mutation to disrupt adhesion and lumen formation |
| GJB1 | Angiogenesis-related pathologies | Overexpression or knockout to modulate gap junction communication |
| DCN | Cancer angiogenesis | Overexpression to inhibit tube formation via thrombospondin-1 |
Diabetes and vascular complications
Endothelial tube formation is impaired in diabetes, contributing to vascular damage and poor angiogenesis. Human umbilical cord-derived mesenchymal stem cells ameliorate blood glucose and protect vascular endothelium from diabetic damage through a paracrine mechanism mediated by MAPK/ERK signaling, highlighting the therapeutic potential of targeting this pathway.
Cancer and tumor angiogenesis
Tumor growth and metastasis depend on angiogenesis, in which endothelial tube formation is a key step. Inhibitors of tube formation, such as decorin via thrombospondin-1, have been investigated for anti-angiogenic therapy. The fungicide ciclopirox inhibits lymphatic endothelial cell tube formation by suppressing VEGFR-3-mediated ERK signaling, suggesting potential applications in cancer and lymphatic disorders.
Lymphatic disorders
Lymphatic endothelial tube formation is essential for lymphangiogenesis. Dysregulation can lead to lymphedema or contribute to tumor lymphangiogenesis. VEGFR-3 signaling is critical, and its inhibition blocks tube formation.
From endothelial tube formation-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Is GJB1 required for endothelial tube formation? | Knockout of GJB1 in endothelial cells followed by tube formation assay |
| Does a specific point mutation in CDH5 disrupt lumen formation? | Point mutation knock-in of CDH5 in endothelial cells |
| Can overexpression of DCN inhibit tumor angiogenesis? | Overexpression of DCN in endothelial or tumor cells |
| What is the role of VEGFR3-ERK signaling in lymphatic tube formation? | Knockout or point mutation of VEGFR3 (FLT4) in lymphatic endothelial cells |
| Does Hedgehog signaling regulate endothelial tube formation in vivo? | Knockout of Shh or Smo in mouse embryos |
| Can tagged knock-in of PECAM1 reveal its dynamics during tube formation? | Tagged knock-in of PECAM1 with fluorescent protein |
How to Study the endothelial tube formation Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Tube formation assay | Ability of endothelial cells to form capillary-like structures | Screening genes or drugs affecting tube formation [1, 5, 6, 7, 8] |
| CRISPR knockout | Loss-of-function effects on tube formation | Determining if a gene is required for tube formation [1, 4, 5, 6, 7, 8] |
| CRISPR knock-in | Effects of specific mutations or tags on tube formation | Studying point mutations or protein localization |
| Western blot | Protein expression and phosphorylation | Analyzing signaling pathways (e.g., ERK, PI3K) [3, 6, 8] |
| Immunofluorescence | Localization of proteins and cytoskeletal changes | Visualizing adhesion complexes and lumen formation |
| Migration assay | Endothelial cell migration capacity | Assessing the role of genes in cell movement [1, 7] |
| In vivo angiogenesis models | Vessel formation in living organisms | Validating findings in physiological context |
| RNA-seq | Transcriptional changes during tube formation | Identifying gene expression programs |
In vitro tube formation assay
The endothelial tube formation assay on Matrigel or collagen is a standard method to assess the ability of endothelial cells to form capillary-like structures. It is used to evaluate the effects of gene knockout, overexpression, or pharmacological inhibitors on tube formation [1, 5, 6, 7, 8].
Genetic manipulation with CRISPR
CRISPR-Cas9 knockout, point mutation knock-in, and overexpression models enable precise interrogation of gene function in endothelial tube formation. These approaches can be combined with tube formation assays to determine causal roles [1, 4, 5, 6, 7, 8].
Signaling pathway analysis
Western blotting, immunoprecipitation, and kinase activity assays are used to dissect signaling pathways such as PI3K, MAPK/ERK, and Hedgehog that regulate tube formation [3, 4, 6, 8].
In vivo models
Mouse models with endothelial-specific gene knockouts or knock-ins, as well as zebrafish, are used to study endothelial tube formation during development and in disease. These models provide physiological context and can validate in vitro findings.
How CRISPR Can Be Used to Study GO:0120331 endothelial tube formation
Knockout
CRISPR knockout of genes such as GJB1, CDH5, or VEGFR3 in endothelial cells can abolish tube formation, demonstrating their essential roles [1, 5, 8]. Knockout models are invaluable for loss-of-function studies to identify required genes.
Point Mutation
Point mutation knock-in using CRISPR can mimic disease-associated mutations in genes like CDH5 or VEGFR3, allowing researchers to study how specific amino acid changes affect endothelial tube formation and signaling [5, 8].
Knock-in
Knock-in of reporter tags (e.g., GFP) into endogenous loci such as PECAM1 enables real-time visualization of protein dynamics during tube formation. Knock-in of mutant alleles can also model human vascular diseases.
Overexpression
CRISPR activation or cDNA overexpression of genes like DCN or GJB1 can enhance or inhibit tube formation, providing gain-of-function insights [1, 7]. Overexpression models are useful for studying pro- or anti-angiogenic factors.
How EDITGENE Supports endothelial tube formation Research
Researchers studying endothelial tube formation-related genes often need to determine whether a candidate gene is causally involved in the process or merely correlated with it. EDITGENE provides a comprehensive suite of CRISPR-based services to enable such causal investigations, from gene knockout to precise point mutations and overexpression, tailored to vascular biology research.
Contact EDITGENE today to design your custom CRISPR model for endothelial tube formation research.
Frequently Asked Questions About endothelial tube formation
What is endothelial tube formation?
Endothelial tube formation is the developmental process pertaining to the initial formation of an endothelial tube, a key step in blood and lymphatic vessel development.
What genes are involved in endothelial tube formation?
Key genes include GJB1 (connexin32), CDH5 (cadherin 5), PECAM1 (CD31), SHH, CXCL12, PIK3CA, DCN, THBS1, VEGFR3 (FLT4), and MAPK1/3 [1, 4, 5, 6, 7, 8].
What is GO:0120331?
GO:0120331 is the Gene Ontology identifier for endothelial tube formation, a biological process.
How is endothelial tube formation studied?
It is studied using in vitro tube formation assays, CRISPR knockout/knock-in models, signaling pathway analysis, and in vivo animal models [1, 4, 5, 6, 7, 8].
What signaling pathways regulate endothelial tube formation?
Hedgehog, PI3K, MAPK/ERK, and VEGFR-3 signaling pathways are key regulators [4, 6, 8].
What diseases are associated with defective endothelial tube formation?
Diabetes, cancer, and lymphatic disorders are associated with dysregulated endothelial tube formation [3, 7, 8].
Can CRISPR be used to study endothelial tube formation?
Yes, CRISPR knockout, knock-in, point mutation, and overexpression models are widely used to dissect gene function in endothelial tube formation [1, 4, 5, 6, 7, 8].
What is the role of connexin32 in endothelial tube formation?
Connexin32 (GJB1) enhances endothelial tube formation and cell migration, likely through gap junction communication.
How does cadherin 5 contribute to tube formation?
Cadherin 5 (CDH5) mediates endothelial cell-cell adhesion and interacts with filamentous actin to support tube formation and lumen formation.
What is the function of VEGFR-3 in tube formation?
VEGFR-3 mediates lymphatic endothelial tube formation through ERK signaling; its inhibition blocks tube formation.
Conclusion
Endothelial tube formation (GO:0120331) is a fundamental developmental process required for vascular and lymphatic system formation. Its dysregulation underlies major human diseases, including diabetes, cancer, and lymphatic disorders. The molecular mechanisms involve a complex interplay of signaling pathways, adhesion molecules, and gap junctions, with key roles for genes such as GJB1, CDH5, PECAM1, SHH, CXCL12, and VEGFR3. CRISPR-based approaches offer powerful tools to dissect these mechanisms and identify therapeutic targets. EDITGENE provides comprehensive CRISPR services to support researchers in advancing this field.
References
- 1. Okamoto T et al.. 2014. Endothelial connexin32 enhances angiogenesis by positively regulating tube formation and cell migration.. Exp Cell Res 321(2):133-41 PMID: 24333598
- 2. Krasnow MA et al.. 2002. Tube morphogenesis.. Trends Cell Biol 12(8):351 PMID: 12191903
- 3. Liu Y et al.. 2022. Human umbilical cord-derived mesenchymal stem cells not only ameliorate blood glucose but also protect vascular endothelium from diabetic damage through a paracrine mechanism mediated by MAPK/ERK signaling.. Stem Cell Res Ther 13(1):258 PMID: 35715841
- 4. Vokes SA et al.. 2004. Hedgehog signaling is essential for endothelial tube formation during vasculogenesis.. Development 131(17):4371-80 PMID: 15294868
- 5. Matsumura T et al.. 1997. Endothelial cell tube formation depends on cadherin 5 and CD31 interactions with filamentous actin.. J Immunol 158(7):3408-16 PMID: 9120301
- 6. Kanda S et al.. 2003. Stromal cell-derived factor-1alpha induces tube-like structure formation of endothelial cells through phosphoinositide 3-kinase.. J Biol Chem 278(1):257-62 PMID: 12414810
- 7. Davies Cde L et al.. 2001. Decorin inhibits endothelial migration and tube-like structure formation: role of thrombospondin-1.. Microvasc Res 62(1):26-42 PMID: 11421658
- 8. Luo Y et al.. 2011. The fungicide ciclopirox inhibits lymphatic endothelial cell tube formation by suppressing VEGFR-3-mediated ERK signaling pathway.. Oncogene 30(18):2098-107 PMID: 21217783
