Aberrant fibrin formation and cross-linking of fibrinogen Nieuwegein, a variant with a shortened Aα-chain, alters endothelial capillary tube formation

Blood ◽  
2001 ◽  
Vol 97 (4) ◽  
pp. 973-980 ◽  
Author(s):  
Annemie Collen ◽  
Annemarie Maas ◽  
Teake Kooistra ◽  
Florea Lupu ◽  
Jos Grimbergen ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Capillary Tube ◽  
Stop Codon ◽  
Tissue Transglutaminase ◽  
Tube Formation ◽  
Factor Xiiia ◽  
Cross Linking ◽  
Molecular Abnormalities ◽  
Fibrin Network

Abstract A congenital dysfibrinogenemia, fibrinogenNieuwegein, was discovered in a young man without any thromboembolic complications or bleeding. A homozygous insertion of a single nucleotide (C) in codon Aα 453 (Pro) introduced a stop codon at position 454, which resulted in the deletion of the carboxyl-terminal segment Aα 454-610. The ensuing unpaired cysteine at Aα 442 generated fibrinogen-albumin complexes of different molecular weights. The molecular abnormalities of fibrinogenNieuwegein led to a delayed clotting and a fibrin network with a low turbidity. Electron microscopy confirmed that thin fibrin bundles were organized in a fine network. The use of fibrinogenNieuwegein-derived fibrin (fibrinNieuwegein) in an in vitro angiogenesis model resulted in a strong reduction of tube formation. The ingrowth of human microvascular endothelial cells (hMVEC) was independent of αvβ3, indicating that the reduced ingrowth is not due to the absence of the RGD-adhesion site at position Aα 572-574. Rather, the altered structure of fibrinNieuwegeinis the cause, since partial normalization of the fibrin network by lowering the pH during polymerization resulted in an increased tube formation. Whereas factor XIIIa further decreased the ingrowth of hMVEC in fibrinNieuwegein, tissue transglutaminase (TG), which is released in areas of vessel injury, did not. This is in line with the absence of the cross-linking site for TG in the α-chains of fibrinogenNieuwegein. In conclusion, this newly discovered congenital dysfibrinogenemia has a delayed clotting time and leads to the formation of an altered fibrin structure, which could not be cross-linked by TG and which is less supportive for ingrowth of endothelial cells.

Abstract P211: N-Acetyl-Seryl-Aspartyl-Lysyl-Proline (Ac-SDKP) Promotes Tube Formation in Human Coronary Artery Endothelial Cells

Hypertension ◽  
2016 ◽  
Vol 68 (suppl_1) ◽  
Author(s):  
Ginette Bordcoch ◽  
Pablo Nakagawa ◽  
Cesar A Romero ◽  
Oscar A Romero
Keyword(s):  
Endothelial Cells ◽  
Coronary Artery ◽  
Capillary Tube ◽  
Tube Formation ◽  
Endothelial Cell Migration ◽  
Tube Length ◽  
Human Coronary Artery ◽  
Double Phase ◽  
Angiogenic Effect

Ac-SDKP is an endogenous peptide with anti-inflammation and anti-fibrotic effects in hypertensive and cardiovascular diseases. It is cleaved from Thymosin β4 (Tβ4) and hydrolyzed by angiotensin converting enzyme (ACE). Ac-SDKP plasma concentration increases after treatment with ACE inhibitors (ACEi) and some of the beneficial effects of ACEi treatment has been ascribed to Ac-SDKP. Ac-SDKP is a mediator of angiogenesis in in-vitro and in-vivo animal models. Ac-SDKP stimulates rodents derived immortalized aortic endothelial cells migration and capillary-like structures formation (tube formation). Similarly, Ac-SDKP increases capillary density after myocardial infarction in rats. The mechanism related to angiogenesis induced by Ac-SDKP is not known. Tβ4 (Ac-SDKP precursor) promotes endothelial cell migration and angiogenesis by the activation of the VEGF/AKT pathway. Our objective is to evaluate the Ac-SDKP pro-angiogenic effect in Human Coronary Artery Endothelial Cells (HCAEC) and the mechanism that regulates the angiogenic effect of Ac-SDKP. HCAEC do not produce VEGF, thus we hypothesize that Ac-SDKP increases VEGF expression in fibroblasts and that indirectly could promote capillary tube formation in endothelial cells. We used primary culture of rat cardiac fibroblast (RCF) and we treated these cells with 10nM Ac-SDKP for 24 hours. VEGF concentration in cell supernatant was measured by ELISA. Cells were starved without serum overnight before the Ac-SDKP treatment. For capillary tube formation assay, HCAEC cells were seeded into matrigel and incubated in presence of 10nM Ac-SDKP for 12 hours, pictures were taken by double phase contrast microscope and tube length was quantified with image J software and the results were expressed as percentage of control. After Ac-SDKP treatment, VEGF concentration did not increase in the supernatant of RCF (control: 0.12±0.07 vs. Ac-SDKP: 0.14±0.09 mg/ml; p=0.7). However, Ac-SDKP treatment induced the development of tube formation in HCAECs by 7±2% respect to control (p=0.037). We conclude that Ac-SDKP induces capillary tube formation not only in rodent but also in human derived endothelial cells. The mechanism by which Ac-SDKP promotes tube formation in HCAECs is still unknown.

Factor XIIIa supports microvascular endothelial cell adhesion and inhibits capillary tube formation in fibrin

Blood ◽  
2000 ◽  
Vol 95 (8) ◽  
pp. 2586-2592
Author(s):  
Susan M. Dallabrida ◽  
Lisa A. Falls ◽  
David H. Farrell
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Endothelial Cell ◽  
Capillary Tube ◽  
Coagulation Factor ◽  
Tube Formation ◽  
Factor Xiiia ◽  
Dependent Manner ◽  
Dose Dependent ◽  
Endothelial Cell Adhesion

Coagulation factor XIIIa is a transglutaminase that catalyzes covalent cross-link formation in fibrin clots. In this report, we demonstrate that factor XIIIa also mediates adhesion of endothelial cells and inhibits capillary tube formation in fibrin. The adhesive activity of factor XIIIa was not dependent on the transglutaminase activity, and did not involve the factor XIIIb-subunits. The adhesion was inhibited by 99% using a combination of monoclonal antibodies directed against integrin vβ3 and β1-containing integrins, and was dependent on Mg2+ or Mn2+. Soluble factor XIIIa also bound to endothelial cells in solution, as detected by flow cytometry. In addition, factor XIIIa inhibited endothelial cell capillary tube formation in fibrin in a dose-dependent manner. Furthermore, the extent of inhibition differed in 2 types of fibrin. The addition of 10 to 100 μg/mL factor XIIIa produced a dose-dependent reduction in capillary tube formation of 60% to 100% in γA/γA fibrin, but only a 10% to 37% decrease in γA/γ′ fibrin. These results show that factor XIIIa supports endothelial cell adhesion in an integrin-dependent manner and inhibits capillary tube formation.

scholarly journals Transglutaminase-mediated oligomerization of the fibrin(ogen) αC domains promotes integrin-dependent cell adhesion and signaling

Blood ◽  
2005 ◽  
Vol 105 (9) ◽  
pp. 3561-3568 ◽  
Author(s):  
Alexey M. Belkin ◽  
Galina Tsurupa ◽  
Evgeny Zemskov ◽  
Yuri Veklich ◽  
John W. Weisel ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Tissue Transglutaminase ◽  
Covalent Modification ◽  
Factor Xiiia ◽  
Cross Linking ◽  
Extracellular Signal ◽  
Integrin Clustering ◽  
Dependent Cell ◽  
Integrin Ligands

AbstractInteractions of endothelial cells with fibrin(ogen) are implicated in inflammation, angiogenesis, and wound healing. Cross-linking of the fibrinogen αC domains with factor XIIIa generates ordered αC oligomers mimicking polymeric arrangement of the αC domains in fibrin. These oligomers and those prepared with tissue transglutaminase were used to establish a mechanism of the αC domain–mediated interaction of fibrin with endothelial cells. Cell adhesion and chemical cross-linking experiments revealed that oligomerization of the αC domains by both transglutaminases significantly increases their RGD (arginyl–glycyl–aspartate)–dependent interaction with endothelial αVβ3 and to a lesser extent with αVβ5 and α5β1 integrins. The oligomerization promotes integrin clustering, thereby increasing cell adhesion, spreading, formation of prominent peripheral focal contacts, and integrin-mediated activation of focal adhesion kinase (FAK) and extracellular signal-regulated kinase (ERK) signaling pathways. The enhanced integrin clustering is likely caused by ordered juxtaposition of RGD-containing integrin-binding sites upon oligomerization of the αC domains and increased affinity of these domains for integrins. Our findings provide new insights into the mechanism of the αC domain–mediated interaction of endothelial cells with fibrin and imply its potential involvement in cell migration. They also suggest a new role for transglutaminases in regulation of integrin-mediated adhesion and signaling via covalent modification of integrin ligands.

scholarly journals Autocrine Role for Interleukin-8 in Bartonella henselae-Induced Angiogenesis

2006 ◽  
Vol 74 (9) ◽  
pp. 5185-5190 ◽  
Author(s):  
Amy M. McCord ◽  
Sandra I. Resto-Ruiz ◽  
Burt E. Anderson
Keyword(s):  
Endothelial Cells ◽  
Capillary Tube ◽  
Bartonella Henselae ◽  
Tube Formation ◽  
Interleukin 8 ◽  
Endothelial Cell Proliferation ◽  
Chemokine Receptor Cxcr2 ◽  
In Vitro Angiogenesis

ABSTRACT The gram-negative bacterium Bartonella henselae is capable of causing angiogenic lesions as a result of infection. Previously, it has been shown that B. henselae infection can result in production of the chemokine interleukin-8 (IL-8). In this study, we demonstrated that monocytes, endothelial cells, and hepatocytes produce IL-8 in response to B. henselae infection. We also investigated the role of IL-8 in B. henselae-induced endothelial cell proliferation and capillary tube formation. Both in vitro angiogenesis assays were IL-8 dependent. B. henselae-mediated inhibition of apoptosis, as indicated by gene expression of Bax and Bcl-2, was also shown to be IL-8 dependent in endothelial cells. Furthermore, infection of endothelial cells with B. henselae stimulated upregulation of the IL-8 chemokine receptor CXCR2. Infection of human endothelial cells by B. henselae resulting in IL-8 production likely plays a central role in the ability of this organism to cause angiogenesis during infection.

Factor XIIIa supports microvascular endothelial cell adhesion and inhibits capillary tube formation in fibrin

Blood ◽  
2000 ◽  
Vol 95 (8) ◽  
pp. 2586-2592 ◽  
Author(s):  
Susan M. Dallabrida ◽  
Lisa A. Falls ◽  
David H. Farrell
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Endothelial Cell ◽  
Capillary Tube ◽  
Coagulation Factor ◽  
Tube Formation ◽  
Factor Xiiia ◽  
Dependent Manner ◽  
Dose Dependent ◽  
Endothelial Cell Adhesion

Abstract Coagulation factor XIIIa is a transglutaminase that catalyzes covalent cross-link formation in fibrin clots. In this report, we demonstrate that factor XIIIa also mediates adhesion of endothelial cells and inhibits capillary tube formation in fibrin. The adhesive activity of factor XIIIa was not dependent on the transglutaminase activity, and did not involve the factor XIIIb-subunits. The adhesion was inhibited by 99% using a combination of monoclonal antibodies directed against integrin vβ3 and β1-containing integrins, and was dependent on Mg2+ or Mn2+. Soluble factor XIIIa also bound to endothelial cells in solution, as detected by flow cytometry. In addition, factor XIIIa inhibited endothelial cell capillary tube formation in fibrin in a dose-dependent manner. Furthermore, the extent of inhibition differed in 2 types of fibrin. The addition of 10 to 100 μg/mL factor XIIIa produced a dose-dependent reduction in capillary tube formation of 60% to 100% in γA/γA fibrin, but only a 10% to 37% decrease in γA/γ′ fibrin. These results show that factor XIIIa supports endothelial cell adhesion in an integrin-dependent manner and inhibits capillary tube formation.

scholarly journals TAFI and Pancreatic Carboxypeptidase B Modulate In Vitro Capillary Tube Formation by Human Microvascular Endothelial Cells

2007 ◽  
Vol 27 (10) ◽  
pp. 2157-2162 ◽  
Author(s):  
Ana H.C. Guimarães ◽  
Nancy Laurens ◽  
Ester M. Weijers ◽  
Pieter Koolwijk ◽  
Victor W.M. van Hinsbergh ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Capillary Tube ◽  
Tube Formation ◽  
Microvascular Endothelial Cells ◽  
Carboxypeptidase B ◽  
Microvascular Endothelial

scholarly journals Applications of Engineering Techniques in Microvasculature Design

2021 ◽  
Vol 8 ◽  
Author(s):  
Aleen Al Halawani ◽  
Ziyu Wang ◽  
Linyang Liu ◽  
Miao Zhang ◽  
Anthony S. Weiss
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Capillary Tube ◽  
Tube Formation ◽  
Engineered Tissue ◽  
Post Implantation

Achieving successful microcirculation in tissue engineered constructs in vitro and in vivo remains a challenge. Engineered tissue must be vascularized in vitro for successful inosculation post-implantation to allow instantaneous perfusion. To achieve this, most engineering techniques rely on engineering channels or pores for guiding angiogenesis and capillary tube formation. However, the chosen materials should also exhibit properties resembling the native extracellular matrix (ECM) in providing mechanical and molecular cues for endothelial cells. This review addresses techniques that can be used in conjunction with matrix-mimicking materials to further advance microvasculature design. These include electrospinning, micropatterning and bioprinting. Other techniques implemented for vascularizing organoids are also considered for their potential to expand on these approaches.

scholarly journals Identification of IGPR-1 as a novel adhesion molecule involved in angiogenesis

2012 ◽  
Vol 23 (9) ◽  
pp. 1646-1656 ◽  
Author(s):  
Nader Rahimi ◽  
Kobra Rezazadeh ◽  
John E. Mahoney ◽  
Edward Hartsough ◽  
Rosana D. Meyer
Keyword(s):  
Endothelial Cells ◽  
Cell Migration ◽  
Capillary Tube ◽  
Cell Aggregation ◽  
Cell Interaction ◽  
Tube Formation ◽  
Fiber Formation ◽  
Actin Stress Fiber ◽  
Cell Cell

Angiogenesis—the growth of new blood vessels from preexisting vessels—is an important physiological process and is considered to play a key role in tumor growth and metastasis. We identified the immunoglobulin-containing and proline-rich receptor-1 (IGPR-1, also called TMIGD2) gene as a novel cell adhesion receptor that is expressed in various human organs and tissues, mainly in cells with epithelium and endothelium origins. IGPR-1 regulates cellular morphology, homophilic cell aggregation, and cell–cell interaction. IGPR-1 activity also modulates actin stress fiber formation and focal adhesion and reduces cell migration. Silencing of expression of IGPR-1 by small interfering RNA (siRNA) and by ectopic overexpression in endothelial cells showed that IGPR-1 regulates capillary tube formation in vitro, and B16F melanoma cells engineered to express IGPR-1 displayed extensive angiogenesis in the mouse Matrigel angiogenesis model. Moreover, IGPR-1, through its proline-rich cytoplasmic domain, associates with multiple Src homology 3 (SH3)–containing signaling proteins, including SH3 protein interacting with Nck (SPIN90/WISH), bullous pemphigoid antigen-1, and calcium channel β2. Silencing of expression of SPIN90/WISH by siRNA in endothelial cells showed that SPIN90/WISH is required for capillary tube formation. These features of IGPR-1 suggest that IGPR-1 is a novel receptor that plays an important role in cell–cell interaction, cell migration, and angiogenesis.

Abstract WMP43: Neurorestorative Therapy of Stroke in T2DM Mice With Exosomes Derived From Brain Endothelial Cells

Stroke ◽  
2017 ◽  
Vol 48 (suppl_1) ◽  
Author(s):  
Jieli Chen ◽  
Chengcheng Cui ◽  
Alex Zacharek ◽  
Poornima Venkat ◽  
Peng Yu ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Capillary Tube ◽  
Artery Occlusion ◽  
Tube Formation ◽  
Vessel Density ◽  
Lesion Volume ◽  
Treatment Groups ◽  
Type Two Diabetes ◽  
Cerebral Artery Occlusion

MicroRNA-126 is primarily expressed in endothelial cells (ECs), and is emerging as a key player in the pathogenesis of type two diabetes (T2DM) induced vascular damage. Stroke in T2DM increases vascular and white matter (WM) damage in the ischemic brain, and has a poor prognosis compared to non-DM stroke. In this study, we investigated the neurorestorative effects of exosomes derived from mouse brain ECs (EC-Exo) as a treatment of stroke in T2DM mice. Methods: Adult male BKS.Cg-m+/+Lepr db /J (T2DM) and db/+ (non-DM) control mice (3-4 m) were subjected to right extraluminal permanent distal middle cerebral artery occlusion (dMCAo), and intravenous injection at 3 days after dMCAo with: 1) PBS; 2) Liposome mimic as vehicle control (3X10 10 ); 3): EC-Exo (3x10 10 ); 4) Knockdown of miR-126 in EC-Exo (miR-126-/-EC-Exo, 3X10 10 ); 5) Knock-in miR-126 in EC-Exo (miR-126+/+EC-Exo, 3X10 10 ). Mice were sacrificed at 28 days after dMCAo. Results: 1) T2DM-dMCAo mice exhibit significantly decreased blood serum, brain tissue and brain-EC miR-126 expression compared to non-DM-dMCAo mice. 2) EC-Exo contain higher levels of miR-126 than exosomes derived from other types of cells (smooth muscle cells, neurons, astrocytes and marrow stromal cells); 3) EC-Exo treatment significantly increases axonal outgrowth and increases capillary tube formation in vitro. Compared to PBS or liposome mimic treatment groups, EC-Exo treatment of stroke in T2DM-dMCAo mice: 4) is safe and does not appear to induce adverse side effects in physiological parameters and does not decrease lesion volume, but significantly improves neurological outcome and cognitive function after dMCAo; 5) significantly increases axon, myelin, vessel density and increases artery diameter in the ischemic boundary zone; 6) significantly increases perfused vessel density measured by FITC-dextran injection. 7) miR-126-/-EC-Exo treatment significantly decreases, but miR-126+/+EC-Exo increases miR-126 expression compared to EC-Exo control. 8) miR-126+/+EC-Exo treatment significantly improves functional outcomes when compared to miR-126-/-EC-Exo, or PBS control. Conclusion: Treatment of stroke with EC-Exo increases miR-126 expression and promotes neurorestorative effects after stroke in T2DM mice.

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