Endothelial cell regulation of matrix metalloproteinases

2005 ◽  
Vol 83 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Tara L Haas
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Matrix Metalloproteinases ◽  
Basement Membrane ◽  
Matrix Proteins ◽  
Regulation Of Transcription ◽  
Cell Type ◽  
Sprouting Angiogenesis

The process of sprouting angiogenesis requires that the endothelial cells degrade the basement membrane matrix and migrate into the interstitial matrix. Matrix metalloproteinases are enzymes capable of cleaving numerous extracellular matrix proteins. Increased production and activity of matrix metalloproteinases in any cell type is associated with a more migratory and invasive phenotype. This paper describes results of recent in-vitro studies of the regulation of transcription and activation of MMP-2 and MT1-MMP in endothelial cells, as well as studies that examined roles of matrix metalloproteinases in activity-induced angiogenesis.Key words: proteolysis, extracellular matrix, angiogenesis, mechanotransduction.

scholarly journals PKM2 regulates endothelial cell junction dynamics and angiogenesis via ATP production

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Jesús Gómez-Escudero ◽  
Cristina Clemente ◽  
Diego García-Weber ◽  
Rebeca Acín-Pérez ◽  
Jaime Millán ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Inflammatory Disease ◽  
Chronic Inflammatory Disease ◽  
Cell Junctions ◽  
Cell Junction ◽  
Collective Migration ◽  
Sprouting Angiogenesis ◽  
Cell Cell

Abstract Angiogenesis, the formation of new blood vessels from pre-existing ones, occurs in pathophysiological contexts such as wound healing, cancer, and chronic inflammatory disease. During sprouting angiogenesis, endothelial tip and stalk cells coordinately remodel their cell-cell junctions to allow collective migration and extension of the sprout while maintaining barrier integrity. All these processes require energy, and the predominant ATP generation route in endothelial cells is glycolysis. However, it remains unclear how ATP reaches the plasma membrane and intercellular junctions. In this study, we demonstrate that the glycolytic enzyme pyruvate kinase 2 (PKM2) is required for sprouting angiogenesis in vitro and in vivo through the regulation of endothelial cell-junction dynamics and collective migration. We show that PKM2-silencing decreases ATP required for proper VE-cadherin internalization/traffic at endothelial cell-cell junctions. Our study provides fresh insight into the role of ATP subcellular compartmentalization in endothelial cells during angiogenesis. Since manipulation of EC glycolysis constitutes a potential therapeutic intervention route, particularly in tumors and chronic inflammatory disease, these findings may help to refine the targeting of endothelial glycolytic activity in disease.

scholarly journals Perturbations in the fibrinolytic pathway abolish cyst formation but not capillary-like organization of cultured murine endothelial cells

Blood ◽  
1994 ◽  
Vol 83 (11) ◽  
pp. 3206-3217 ◽  
Author(s):  
N Dubois-Stringfellow ◽  
A Jonczyk ◽  
VL Bautch
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Basement Membrane ◽  
Organizational Behavior ◽  
Cell Lines ◽  
Primary Cultures ◽  
Cyst Formation ◽  
Fibrin Gels

Abstract Fibrinolytic activity and its relation to morphogenesis was investigated in several transformed murine endothelial cell lines and primary cultures of endothelial cells. Two in vitro systems, fibrin gels and Matrigel (Collaborative Research, Bedford, MA), were used. Fibrin gels model a fibrin-rich extracellular matrix that frequently supports neovascularization in vivo, and Matrigel models the basement membrane surrounding quiescent endothelial cells in vivo. The transformed endothelial cell lines have higher levels of plasminogen activator (PA) mRNA than primary cultures of endothelial cells, and an increased PA-mediated proteolytic activity was correlated with formation of cysts in fibrin gels. Addition of neutralizing anti- urokinase antibodies, plasminogen depletion, or addition of a plasmin inhibitor prevented cyst formation. Addition of plasminogen restored the ability to form cysts in the plasminogen-depleted system. Normal endothelial cells organized into capillary-like structures in fibrin gels regardless of manipulations affecting the fibrinolytic pathway. In Matrigel, both transformed and primary cultures of endothelial cells rapidly formed a capillary-like network that was not affected by plasminogen depletion or addition of plasmin inhibitors. Thus, elements of the fibrinolytic pathway necessary for cyst formation are not critical in capillary-like structure formation on a reconstituted basement membrane. These results suggest that plasmin is essential for hemangioma formation but is not critical to the organizational behavior of normal endothelial cells.

scholarly journals Aberrant production of extracellular matrix proteins and dysfunction in kidney endothelial cells with a short duration of diabetes

2013 ◽  
Vol 304 (1) ◽  
pp. F19-F30 ◽  
Author(s):  
Cathy Grutzmacher ◽  
SunYoung Park ◽  
Yun Zhao ◽  
Margaret E. Morrison ◽  
Nader Sheibani ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Diabetic Nephropathy ◽  
Endothelial Cell ◽  
Short Duration ◽  
Cell Function ◽  
Matrix Proteins ◽  
Endothelial Cell Function ◽  
Duration Of Diabetes ◽  
Akita Mice

Diabetic nephropathy is the most common cause of end-stage renal disease and is a major risk factor for cardiovascular disease. In the United States, microvascular complications during diabetic nephropathy contribute to high morbidity and mortality rates. However, the cell-autonomous impact of diabetes on kidney endothelial cell function requires further investigation. Male Akita/+ [autosomal dominant mutation in the insulin II gene (Ins2)] mice reproducibly develop diabetes by 4 wk of age. Here, we examined the impact a short duration of diabetes had on kidney endothelial cell function. Kidney endothelial cells were prepared from nondiabetic and diabetic mice (4 wk of diabetes) to delineate the early changes in endothelial cell function. Kidney endothelial cells from Akita/+ mice following 4 wk of diabetes demonstrated aberrant expression of extracellular matrix proteins including decreased osteopontin and increased fibronectin expression which correlated with increased α5-integrin expression. These changes were associated with the attenuation of migration and capillary morphogenesis. Kidney endothelial cells from Akita/+ mice had decreased VEGF levels but increased levels of endothelial nitric oxide synthase(eNOS) and NO, suggesting uncoupling of VEGF-mediated NO production. Knocking down eNOS expression in Akita/+ kidney endothelial cells increased VEGF expression, endothelial cell migration, and capillary morphogenesis. Furthermore, attenuation of sprouting angiogenesis of aortas from Akita/+ mice with 8 wk of diabetes was restored in the presence of the antioxidant N-acetylcysteine. These studies demonstrate that aberrant endothelial cell function with a short duration of diabetes may set the stage for vascular dysfunction and rarefaction at later stages of diabetes.

scholarly journals Perturbations in the fibrinolytic pathway abolish cyst formation but not capillary-like organization of cultured murine endothelial cells

Blood ◽  
1994 ◽  
Vol 83 (11) ◽  
pp. 3206-3217 ◽  
Author(s):  
N Dubois-Stringfellow ◽  
A Jonczyk ◽  
VL Bautch
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Basement Membrane ◽  
Organizational Behavior ◽  
Cell Lines ◽  
Primary Cultures ◽  
Cyst Formation ◽  
Fibrin Gels

Fibrinolytic activity and its relation to morphogenesis was investigated in several transformed murine endothelial cell lines and primary cultures of endothelial cells. Two in vitro systems, fibrin gels and Matrigel (Collaborative Research, Bedford, MA), were used. Fibrin gels model a fibrin-rich extracellular matrix that frequently supports neovascularization in vivo, and Matrigel models the basement membrane surrounding quiescent endothelial cells in vivo. The transformed endothelial cell lines have higher levels of plasminogen activator (PA) mRNA than primary cultures of endothelial cells, and an increased PA-mediated proteolytic activity was correlated with formation of cysts in fibrin gels. Addition of neutralizing anti- urokinase antibodies, plasminogen depletion, or addition of a plasmin inhibitor prevented cyst formation. Addition of plasminogen restored the ability to form cysts in the plasminogen-depleted system. Normal endothelial cells organized into capillary-like structures in fibrin gels regardless of manipulations affecting the fibrinolytic pathway. In Matrigel, both transformed and primary cultures of endothelial cells rapidly formed a capillary-like network that was not affected by plasminogen depletion or addition of plasmin inhibitors. Thus, elements of the fibrinolytic pathway necessary for cyst formation are not critical in capillary-like structure formation on a reconstituted basement membrane. These results suggest that plasmin is essential for hemangioma formation but is not critical to the organizational behavior of normal endothelial cells.

scholarly journals Equine Endothelial Cells Support Productive Infection of Equine Infectious Anemia Virus

1998 ◽  
Vol 72 (11) ◽  
pp. 9291-9297 ◽  
Author(s):  
Wendy Maury ◽  
J. Lindsay Oaks ◽  
Sarahann Bradley
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Equine Infectious Anemia Virus ◽  
High Titer ◽  
Productive Infection ◽  
Cell Type ◽  
Cell Infection ◽  
Equine Infectious Anemia ◽  
Anemia Virus

ABSTRACT Previous cell infectivity studies have demonstrated that the lentivirus equine infectious anemia virus (EIAV) infects tissue macrophages in vivo and in vitro. In addition, some strains of EIAV replicate to high titer in vitro in equine fibroblasts and fibroblast cell lines. Here we report a new cell type, macrovascular endothelial cells, that is infectible with EIAV. We tested the ability of EIAV to infect purified endothelial cells isolated from equine umbilical cords and renal arteries. Infectivity was detected by cell supernatant reverse transcriptase positivity, EIAV antigen positivity within individual cells, and the detection of viral RNA by in situ hybridization. Virus could rapidly spread through the endothelial cultures, and the supernatants of infected cultures contained high titers of infectious virus. There was no demonstrable cell killing in infected cultures. Three of four strains of EIAV that were tested replicated in these cultures, including MA-1, a fibroblast-tropic strain, Th.1, a macrophage-tropic strain, and WSU5, a strain that is fibroblast tropic and can cause disease. Finally, upon necropsy of a WSU5-infected horse 4 years postinfection, EIAV-positive endothelial cells were detected in outgrowths of renal artery cultures. These findings identify a new cell type that is infectible with EIAV. The role of endothelial cell infection in the course of equine infectious anemia is currently unknown, but endothelial cell infection may be involved in the edema that can be associated with infection. Furthermore, the ability of EIAV to persistently infect endothelial cultures and the presence of virus in endothelial cells from a long-term carrier suggest that this cell type can serve as a reservoir for the virus during subclinical phases of infection.

Tensional forces in fibrillar extracellular matrices control directional capillary sprouting

1999 ◽  
Vol 112 (19) ◽  
pp. 3249-3258 ◽  
Author(s):  
T. Korff ◽  
H.G. Augustin
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Three Dimensional ◽  
Type I ◽  
Cellular Delivery ◽  
Form Complex ◽  
Cell Spheroid ◽  
In Vitro Angiogenesis

During angiogenesis, anastomosing capillary sprouts align to form complex three-dimensional networks of new blood vessels. Using an endothelial cell spheroid model that was developed to study endothelial cell differentiation processes, we have devised a novel collagen gel-based three-dimensional in vitro angiogenesis assay. In this assay, cell number-defined, gel-embedded endothelial cell spheroids act as a cellular delivery device, which serves as a focal starting point for the sprouting of lumenized capillary-like structures that can be induced to form complex anastomosing networks. Formation of capillary anastomoses is associated with tensional remodeling of the collagen matrix and directional sprouting of outgrowing capillaries towards each other. To analyze whether directional sprouting is dependent on cytokine gradients or on endothelial cell-derived tractional forces transduced through the extracellular matrix, we designed a matrix tension generator that enables the application of defined tensional forces on the extracellular matrix. Using this matrix tension generator, causal evidence is presented that tensional forces on a fibrillar extracellular matrix such as type I collagen, but not fibrin, are sufficient to guide directional outgrowth of endothelial cells. RGD peptides but not control RAD peptides disrupted the integrity of sprouting capillary-like structures and induced detachment of outgrowing endothelial cells cultured on top of collagen gels, but did not inhibit primary outgrowth of endothelial cells. The data establish the endothelial cell spheroid-based three-dimensional angiogenesis technique as a standardized, highly reproducible quantitative assay for in vitro angiogenesis studies and demonstrate that integrin-dependent matrix tensional forces control directional capillary sprouting and network formation.

Improved Adhesion and Proliferation of Human Endothelial Cells on Polyethylene Precoated with Monoclonal Antibodies Directed against Cell Membrane Antigens and Extracellular Matrix Proteins

1991 ◽  
Vol 66 (06) ◽  
pp. 715-724 ◽  
Author(s):  
Albert Dekker ◽  
André A Poot ◽  
Jan A van Mourik ◽  
Martin P A Workel ◽  
Tom Beugeling ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Monoclonal Antibodies ◽  
Endothelial Cell ◽  
Platelet Reactivity ◽  
Extracellular Matrix Proteins ◽  
Matrix Proteins ◽  
Specific Monoclonal Antibody ◽  
Human Endothelial Cells ◽  
Membrane Antigens

SummaryEndothelial cell seeding may improve the patency of synthetic vascular grafts provided that platelet reactivity of non-endothelialized sites is not increased. We have investigated if surface-adsorbed monoclonal antibodies directed against endothelial cell membrane proteins and against extracellular matrix proteins promote the adhesion and proliferation of cultured human endothelial cells, without causing platelet deposition at non-endothelialized sites. Adhesion of endothelial cells onto polyethylene coated with monoclonal antibodies directed against endothelial cell-specific membrane antigens, integrin receptors and glycoprotein CD31 was equal to or higher than adhesion onto fibronectin-coated polyethylene. Endothelial cells did not proliferate on these surface-adsorbed antibodies. However, pre-coating of polyethylene with mixtures of endothelial cell-specific monoclonal antibodies and monoclonal antibodies directed against fibronectin or von Willebrand factor, resulted in relatively high adhesion and optimal proliferation. Platelet reactivity of the polyethylene surface was found to significantly increase after adsorption of fibronectin, endothelial cell-specific monoclonal antibody or its Fc fragments. In contrast, adsorption of F(ab')2 fragments of endothelial cell-specific monoclonal antibody did not promote platelet deposition. Therefore, it is concluded that coating of vascular graft materials with mixtures of F(ab')2 fragments of monoclonal antibodies specifically directed against endothelial cells and against extracellular matrix proteins may be an effective way to both promote the growth of seeded endothelial cells and limit platelet-graft interaction.

Enforced expression of tissue inhibitor of matrix metalloproteinase-3 affects functional capillary morphogenesis and inhibits tumor growth in a murine tumor model

Blood ◽  
2002 ◽  
Vol 100 (9) ◽  
pp. 3361-3368 ◽  
Author(s):  
William W. Spurbeck ◽  
Catherine Y. C. Ng ◽  
Ted S. Strom ◽  
Elio F. Vanin ◽  
Andrew M. Davidoff
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Matrix Metalloproteinases ◽  
Tumor Growth ◽  
Tumor Cells ◽  
Severe Combined Immunodeficiency ◽  
Melanoma Tumor ◽  
Vitro Assay ◽  
Capillary Morphogenesis

Abstract Homeostasis of the extracellular matrix is a delicate balance between degradation and remodeling, the balance being maintained by the interaction of activated matrix metalloproteinases (MMPs) and specific tissue inhibitors of matrix metalloproteinases (TIMPs). Up-regulation of MMP activity, favoring proteolytic degradation of the basement membrane and extracellular matrix, has been linked to tumor growth and metastasis, as well as tumor-associated angiogenesis, whereas inhibition of MMP activity appears to restrict these processes. We have used retroviral-mediated gene delivery to effect sustained autocrine expression of TIMP-3 in murine neuroblastoma and melanoma tumor cells in order to further examine the ability of TIMPs to inhibit angiogenesis in vivo. Growth of both histologic types of gene-modified tumor cells in severe combined immunodeficiency (SCID) mice was significantly restricted when compared with controls. Grossly, these tumors were small and had few feeding vessels. Histologic evaluation revealed that although tumors overexpressing TIMP-3 had an increased number of CD31+endothelial cells, these endothelial cells had not formed functional tubules, as evidenced by decreased vessel continuity and minimal pericyte recruitment. This effect appears to be mediated, in part, by decreased expression of vascular endothelial (VE)–cadherin by endothelial cells in the presence of TIMP-3 as seen both in an in vitro assay and in TIMP-3–overexpressing tumors. Taken together, these results demonstrate that overexpression of TIMP-3 can inhibit angiogenesis and associated tumor growth, and that the antiangiogenic effects of TIMP-3 appear to be mediated through the inhibition of functional capillary morphogenesis.

Abstract 20152: Multimerin2 Influences Vascularization Through Modulation of the Extracellular Matrix

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Petra E Burgisser ◽  
Dennie Tempel ◽  
Caroline Cheng ◽  
Gerard Pasterkamp ◽  
Henricus Duckers
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Tube Formation ◽  
Endothelial Cell Proliferation ◽  
Mouse Development ◽  
New Genes ◽  
A Genome ◽  
Extracellular Matrix Components

Background: Vascular development is crucial in normal physiology and pathophysiology, and is in part orchestrated by a dynamic balance between extracellular matrix (ECM) and endothelial cells. The molecular mechanisms that govern this balance and as such play a role in the formation of new blood vessels remain largely unknown. In order to identify new genes involved in blood vessel formation, we performed a genome-wide gene expression array analysis in Flk1+ angioblasts during mouse development using a commercial platform (Affymetrix). Multimerin2 (Mmrn2) expression was found to be upregulated in angioblasts and endothelial cells and in the developing vasculature of zebrafish and mice. Methods & Results: Subsequent in vitro studies using an adenoviral construct to induce Mmrn2 overexpression showed a negative effect on angiogenesis. Mmrn2 overexpression resulted in a reduction of 2D tube formation capacity of human aortic endothelial cells (HAECs), as seen in a decrease in the number of junctions and tubules (P<0.05, N=3). Another observed effect after Mmrn2 overexpression was a G0/G1 cell cycle arrest in HAECs (P<0.001, N=8) and decreased migration of the cells in a Boyden chamber assay (P<0.05, N=10). Further in vitro experiments, using immunofluorescence, showed Mmrn2 was detected outside of HAECs, were it co-localized with the extracellular matrix components perlecan, laminin and fibronectin. Direct interaction with these proteins was confirmed using a combination of co-immunoprecipitation followed by proteomics (LC-MS), and western blot to identify possible binding partners. Interestingly, overexpression of Mmrn2 also resulted in enhanced expression of these extracellular components, suggesting that Mmrn2 changes the phenotype from a more proliferative endothelial cell towards a more productive endothelial cell. Conclusions: These findings suggest that Mmrn2 is an abundant and important extracellular matrix protein, secreted by endothelial cells, which influences the expression of other ECM proteins such as fibronectin, laminin and perlecan. We hypothesize that Mmrn2 switches the phenotype of the endothelial cell and subsequently diminishes endothelial cell proliferation, migration and overall tube formation.

Sign in / Sign up

Export Citation Format

Share Document