Human endothelial gelatinases and angiogenesis

doi:10.1016/S1357-2725(01)00007-3

The International Journal of Biochemistry & Cell Biology

Volume 33, Issue 10, October 2001, Pages 960-970

https://doi.org/10.1016/S1357-2725(01)00007-3 Get rights and content

Abstract

Endothelial cell invasion is an essential event during angiogenesis (formation of new blood vessels). The process involves the degradation of the basement membrane and the underlying interstitium. The matrix metalloproteinase (MMP) family is considered to be primarily responsible for matrix degradation. Two members of the family, gelatinase A and B play an important role in angiogenesis. This review outlines recent findings on their regulation in human endothelial cells. Latent gelatinase B is secreted from endothelial cells. This enzyme can also accumulate in the cytosol as an active enzyme, free of TIMP-1. In contrast, latent galatinase A is constitutively secreted from the cells. Unlike other MMPs, gelatinase A activation occurs on the cell membrane and is mediated by MT1-MMP. A number of physiological activators have recently been described. These include thrombin and activated protein C, both of which activate gelatinase A independent of the MT1-MMP pathway. These new findings may lead to therapeutic interventions for the treatment of angiogenic-dependent diseases such as cancer and arthritis.

Introduction

Angiogenesis is a process by which new blood vessels develop from the existing microvascular bed. Physiological angiogenesis, which occurs in reproduction, placental development and wound repair, usually occurs in short bursts and is self-limiting. In contrast, pathological angiogenesis, which occurs in a number of diseases such as solid tumours and rheumatoid arthritis, often persists indefinitely [1], [2]. Folkman was the first to propose that vascularization of tumour growth is essential for its survival [3].

Angiogenesis occurs by a series of sequential steps. In response to angiogenic stimuli, endothelial cells that line the existing microvessels degrade their basement membrane by secreting proteolytic enzymes including the matrix metalloproteinases (MMPs) and serine proteases [4]. The cells then migrate through the degraded basement membrane and continue to break down the interstitial stroma as they move. The migrating endothelial cells at the tip of sprout do not usually divide, whereas the trailing cells at the base of the new vessel undergo proliferation. The endothelium then aligns in a bipolar fashion to form a lumen. The newly formed hollow sprouts anastomose with each other to form a capillary through which, blood flows.

Recent findings have indicated that MMPs, particularly gelatinase A and B, play a central role during angiogenesis. Some characteristics of these enzymes are shown in Table 1. This review highlights recent advances in the regulation of these two enzymes in human endothelial cells as well as their role in angiogenesis.

Section snippets

Matrix metalloproteinases (MMPs)

MMPs are a family of enzymes that play a central role in ECM turnover and remodelling based on their ability to hydrolyze major protein components of the ECM [5]. They are secreted as inactive proenzymes that require activation by the removal of the propeptide, revealing the Zn-binding active site. MMPs are active at neutral pH and require Ca²⁺ for full activity. They are specifically inhibited by the tissue inhibitors of MMPs (TIMPs). There are four TIMPs, of which, TIMP-1 appears to be the

Gelatinase are vital during angiogenesis

Both gelatinase A and B are well known for their ability to degrade collagens present in the vascular basement membrane [29]. The gelatinases also assist the collagenases in the degradation of the interstitium. Gelatinase A can breakdown the interstitial components by directly cleaving type I collagen at a rate similar to that of interstitial collagenase [30]. Crabbe et al. [31] have reported that active gelatinase A can promote the activation of latent interstitial collagenase. The latter

Human gelatinase B

Human gelatinase B is synthesised as a polypeptide of M_r of 78 426 [38]. The proenzyme has a M_r of 92 kD, but migrates as an 88 kD protein under non-reducing conditions on SDS-polyacrylamide gel. The secreted gelatinase B is heavily glycosylated due to the presence of three N-linked glycosylation sites and several O-linked glycosylation sites, thus accounting for the extra mass of the mature enzyme. The structure of the human gelatinase B gene has been determined by Huhtala et al. [39]. The

Regulation of gelatinase B in endothelial cells

Human progelatinase B is secreted from endothelial cells in response to the tumour promoting chemical, phorbol myristate acetate (PMA) [40]. There have been few reports on the effects of cytokines or angiogenic/growth factors on gelatinase B secretion by human endothelial cells. Hanemaaijer et al. [41] have shown that TNF-α can enhance the effect of PMA, but it does not stimulate gelatinase B synthesis by endothelial cells when used alone. Marc et al. [42] reported that activated CD40 T cells

Human gelatinase A

Human gelatinase A has a M_r of 72 kD (migrates as a 66 kD protein under non-reducing conditions) on SDS-polyacrylamide gel, in agreement with its amino acid sequence. Although gelatinase A has a similar substrate specificity to that of gelatinase B, it is differently regulated at both transcriptional and extracellular levels [45]. Unlike other MMPs, the promoter of gelatinase A gene lacks the TRE sequence as well as the known transactivator sequences, AP-1 and PEA-3 [46]. This may explain the

Regulation of gelatinase A activation in endothelial cells

Human endothelial cells constitutively secrete latent gelatinase A. The expression of the latent enzyme is not upregulated by PMA [50], [51], although Hanemaaijer et al. [41] have shown that stimulation of umbilical vein endothelial cells (HUVE) with PMA resulted in the secretion of the two active forms (64 and 62 kD) of gelatinase A. Similarly, progelayinase A activation induced by PMA has been reported in neonatal foreskin microvascular endothelial cells [52]. Lewalle et al. [50] demonstrated

Current model of the role of gelatinases in angiogenesis

Based on recent findings, a model to explain the role of gelatinases during angiogenesis, in diseases such as cancer and arthritis, is described below and is depicted diagrammatically in Fig. 2. Endothelial cells continually secrete latent gelatinase A under basal conditions in vitro. In vivo, gelatinase A has been found to be strongly expressed by some endothelial cell types, including in human glioblastomas [63]. Thrombin, which is present at high levels in angiogenic situations such as

Conclusions and future considerations

It has become apparent in recent years that gelatinases are vital for angiogenesis. The recent discoveries revealing mechanisms of gelatinase regulation in human endothelial cells have greatly contributed to the understanding of their role in angiogenesis. However, a number of questions remain unanswered. What triggers the release of gelatinase B from secretory vessels in the endothelial cell? How is intracellular gelatinase B activated? Of the two known pathways involved in the activation of

References (71)

T.E. Cawston
Metalloproteinase inhibitors and the prevention of connective tissue breakdown
Pharm. Therapeu.
(1996)
S.K. Chandler et al.
Matrix metalloproteinases degrade myelin basic protein
Neurosci. Lett.
(1995)
K. Shofuda et al.
Expression of three membrane-type matrix metalloproteinase (MT-MMPs) in rat vascular smooth muscle cells and characterization of MT3-MMP with and without transmembrane domain
J. Biol. Chem.
(1997)
R. Montesano et al.
Tumor-promoting phorbol esters induce angiogenesis in vitro
Cell
(1985)
C. Fisher et al.
Interstitial collagenase is required for angiogenesis in vitro
Dev. Biol.
(1994)
G. Murphy et al.
Gelatinases A and B
Methods Enzymol.
(1995)
R.T. Aimes et al.
Matrix metalloproteinase-2 is an interstitial collagenase
J. Biol. Chem.
(1995)
P.C. Brooks et al.
Disruption of angiogenesis by PEX, a noncatalytic metalloproteinase fragment with integrin binding activity
Cell
(1998)
P.C. Brooks et al.
Localization of matrix metalloproteinase MMP-2 to the surface of invasive cells by interaction with integrin α_vβ₃
Cell
(1996)
T.H. Vu et al.
MMP-9/gelatinase B is a key regulator of growth plate angiogenesis and apoptosis of hypertrophic chondrocytes
Cell
(1998)

L.A. Cornelius et al.

Human dermal microvascular endothelial cells produce matrix metalloproteinases in response to angiogenic factors and migration

J. Invest. Dermatol.

(1995)

F. Mach et al.

Lymphocytes induce endothelial cell matrix metalloproteinase expression by a CD40L-dependent mechanism — implications for tubule formation

Am. J. Path.

(1999)

M. Nguyen et al.

Active and tissue inhibitor of matrix metalloproteinase-free gelatinase B accumulates within human microvascular endothelial vesicles

J. Biol. Chem.

(1998)

G.S. Butler et al.

The TIMP-2 membrane type 1 metalloproteinase receptor regulates the concentration and efficient activation of progelatinase A — a kinetic study

J. Biol. Chem.

(1998)

V.T. Chan et al.

Membrane-type matrix metalloproteinases in human dermal microvascular endothelial cells: expression and morphogenetic correlation

J. Invest. Dermatol.

(1998)

M. Nguyen et al.

Three-dimensional collagen matrices induce delayed but sustained activation of gelatinase A in human endothelial cells via MT1-MMP

Inter. J. Biochem. Cell Biol.

(2000)

M. Nguyen et al.

Activated protein C directly activates human endothelial gelatinase A

J. Biol. Chem.

(2000)

H. Wang et al.

Hepatocyte growth factor enhances MMP activity in human endothelial cells

Biochem. Biophys. Res. Commun.

(2000)

H.G. Kim et al.

Lipopolysaccharide activates matrix metalloproteinase-2 in endothelial cells through an NF-kB-dependent pathway

Biochem. Biophys. Res. Commun.

(2000)

G. Vince et al.

Heterogenenous regional expression patterns of matrix metalloproteinases in human malignant gliomas

Inter. J. Dev. Neuro.

(1999)

J. Folkman

Angiogenesis in cancer, vascular, rheumatoid and other disease

Nature Med.

(1995)

L. Liotta, P.S. Steeg, W.G. Stetler Stevenson, Cancer metastasis and angiogenesis: an imbalance of positive and...

J. Folkman

Tumour angiogenesis: therapeutic implications

New Eng. J. Med.

(1971)

P.N. Furness

Basement membrane synthesis and degradation

J. Pathol.

(1997)

H. Birkedalhansen

Role of matrix metalloproteinases in human periodontal diseases

J. Periodon.

(1993)

T.A. Butler et al.

Inhibition of ovulation in the perfused rat ovary by the synthetic collagenase inhibitor SC 44463

Biol. Reprod.

(1991)

C.H. Graham et al.

Mechanism of control of trophoblast invasion in situ

J. Cell. Phys.

(1991)

S. Gack et al.

Expression of interstitial collagenase during skeletal development of the mouse is restricted to osteoblast-like cells and hypertrophic chondrocytes

Cell Growth Differ.

(1995)

L.R. Lund et al.

Two distinct phases of apoptosis in mammary gland involution: proteinase-independent and -dependent pathways

Development

(1996)

S. Stetler

Matrix metalloproteinases in angiogenesis: a moving target for therapeutic intervention

J. Clin. Invest.

(1999)

D.E. Kleiner et al.

Matrix metalloproteinases and metastasis

Cancer Chemo. Pharmacol.

(1999)

H.I. Park, J. Ni, F.E. Gerkema, D. Liu, V.E. Belozerov, Q.X.A. Sang, Identification and characterization of human...

J.F.J. Woessner

Matrix metalloproteinases and their inhibitors in connective tissue remodelling

FASEB J.

(1991)

L.M. Matrisian

The matrix-degrading metalloproteinases

Bioessays

(1992)

H. Sato et al.

Matrix metalloproteinase expressed on the surface of invasive tumour cells

Nature

(1994)

Cited by (267)

Skin wound healing: The critical role of angiogenesis
2022, Biomaterials for Vasculogenesis and Angiogenesis
Management of skin wounds is of great clinical importance. Inducing the formation of new blood vessels (angiogenesis) can accelerate the healing process in patients suffering from acute and chronic wounds (e.g., burns and diabetic ulcers). It is currently accepted that angiogenesis plays a substantial role in all four overlapping phases of normal wound healing, that is, hemostasis, inflammation, proliferation, and remodeling. Many experimental studies have focused on the design and development of skin replacements capable of stimulating neovascularization within the skin defect regions. The use of stem/progenitor cells and bioactive molecules (e.g., growth factors and cytokines) in combination with three-dimensional (3D) scaffolds are among the most promising applied strategies for promoting angiogenesis, and subsequently accelerating wound healing. Each approach has its own pros and cons for managing skin wounds. In the present chapter, we first describe the physiology of skin tissue, and the diseases and disorders which affect wound healing. We then summarize current therapies with a focus on tissue-engineering (TE) approaches. The critical role of proangiogenic strategies are discussed to highlight their importance for future studies
In vitro angiogenesis inhibition with selective compounds targeting the key glycolytic enzyme PFKFB3
2021, Pharmacological Research
Abnormal glycolytic metabolism contributes to angiogenic sprouting involved in atherogenesis. We investigated the potential anti-angiogenic properties of specific 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (PFKFB3) inhibitors in endothelial cells (ECs). ECs were treated with PFKFB3 inhibitors (named PA-1 and PA-2) and their effects on metabolic and functional characteristics of ECs were investigated. The anti-glycolytic compound 3-(pyridinyl)− 1-(4-pyridinyl)− 2-propen-1-one (3PO) was used as reference compound. PFKFB3 expression and activity (IC₅₀ about 3–21 nM) was inhibited upon treatment with both compounds. Glucose uptake and lactate export were measured using commercial assays and showed a partial reduction up to 40%. PFKFB3 inhibition increased intracellular lactate accumulation, and reduced expression of monocarboxylate transporters-1 (MCT1) and MCT4. Furthermore, endothelial cell migration and proliferation assays demonstrated significant reduction upon treatment with both compounds. Matrix- metalloproteinase (MMP) activity, measured by gelatin zymography, and expression was significantly reduced (up to 25%). In addition, PA compounds downregulated the expression of VCAM-1, VE-cadherin, VEGFa, VEGFR2, TGF-β, and IL-1β, in inflamed ECs. Finally, PA-1 and PA-2 treatment impaired the formation of angiogenic sprouts measured by both morphogenesis and spheroid-based angiogenesis assays. Our data demonstrate that the anti-glycolytic PA compounds may affect several steps involved in angiogenesis. Targeting the key glycolytic enzyme PFKFB3 might represent an attractive therapeutic strategy to improve the efficacy of cancer treatments, or to be applied in other pathologies where angiogenesis is a detrimental factor.
Angiogenesis: Aspects in wound healing
2021, Endothelial Signaling in Vascular Dysfunction and Disease: From Bench to Bedside
The process of angiogenesis plays a dominant role in effective repairing of wound. In growing tissues freshly developed vessels of blood contribute in development of granulation tissue provisionally besides provide nutrition in addition to oxygen for filling the wound space. Angiogenesis is a complex mechanism that is highly regulated by both serum and underlying extracellular matrix (ECM) signals, in response to tissue injury. The utmost vital angiogenic cytokines in wound angiogenesis include vascular endothelial growth factor, angiopoietins, fibroblastic growth factor, and transforming growth factor-β. Proliferation of new capillaries occurs via a cascade of biological events. In this topic we have summarized angiogenesis, growth factors implicated in the process of angiogenesis, and phases of angiogenesis process in wound healing along with nanocarriers that are intended for promoting angiogenesis.
Self-assembled VEGF-R2 targeting DNA aptamer-collagen fibers stimulate an angiogenic-like endothelial cell phenotype
2021, Materials Science and Engineering C
Citation Excerpt :
Lastly, we measured secreted matrix metalloproteinase-2 (MMP-2), also known as gelatinase A and found that similar to ANGPT-2, there was a statistically significant increase in secreted MMP-2 by endothelial cells cultured on div-VEGF-R2 NACC fibers (Fig. 7). Matrix metalloproteinase-2 secretion is stimulated in endothelial cells following their exposure type I collagen [58]. It acts to degrade type IV collagen within the endothelial cell basement membrane– a necessary step in angiogenesis [42].
Vascularization of engineered tissue is one of the hallmark challenges of tissue engineering. Leveraging self-assembled nucleic acid-collagen complexes (NACCs), we mixed a VEGF-R2 targeting aptamer or its receptor agonist divalent assembly with type I collagen to assemble NACC microfibers. Human umbilical vein endothelial cells (HUVECs) quickly remodeled these fibers into tubulogenic-like structures over 48 h. Moreover, NACCs made with the receptor agonist divalent aptamer assembly promoted enhanced expression of von Willebrand factor (vWF), angiopoietin-2 (ANGPT-2), and matrix metalloproteinase-2 (MMP-2) by HUVECs as measured by either immunocytochemistry or ELISA. The findings suggest, endothelial cell phenotype was directed by both biochemical cues afforded by the agonist behavior of the divalent aptamer assembly as well as by the biophysical cues afforded by the fibrous topography. Collectively, these results support the development of an angiogenic endothelial cell phenotype stimulated by the VEGF-R2 agonist NACC fibers. Thus, the combination of engineered DNA aptamer nanotechnology and DNA-collagen complexation phenomena is a promising biofunctional natural scaffold material system for tissue engineering and regenerative medicine applications.
Immunohistochemical Profiles of Matrix Metalloproteinases and Vascular Endothelial Growth Factor Overexpression in the Antoni B Area of Vestibular Schwannomas
2020, World Neurosurgery
Citation Excerpt :
Endothelial cells shed membrane vesicles containing MMP-2, MMP-9, and MMP-14.30 MMP-2 and MMP-9 (gelatinases A and B) can preferentially cleave collagen type IV, a major component of basement membrane.31 In particular, MMP-14 can degrade several ECM components such as fibrillar collagens, fibronectin, vitronectin, fibrin, and gelatin.32
To evaluate the clinical manifestations of cystic vestibular schwannomas (VSs), investigate the immunohistochemical profiles of matrix metalloproteinases (MMPs) and vascular endothelial growth factor (VEGF) expression in Antoni A and B areas, and speculate the pathogenesis of cystic formation and intratumoral hemorrhage.
Clinical features and outcomes of 24 cases of cystic VSs and 38 cases of solid VSs were retrospectively compared. Immunohistochemical studies were conducted to evaluate the characteristics of MMPs and VEGF in cystic and solid VSs.
The tumor size was 38.92 ± 1.86 mm and 31.95 ± 1.74 mm in the cystic and solid VSs group, respectively (P = 0.011). Cystic VSs were rich in the Antoni B area. MMP-9 expression was low in the Antoni A and B areas. MMP-2 was moderately expressed. No significant difference in MMP-2 expression existed between the Antoni A and B areas (P > 0.05). VEGF and MMP-14 expression were moderate in the Antoni A area and intense in the Antoni B area, and the expression of both was significantly greater in the Antoni B area than in the Antoni A area (P < 0.001).
MMP-14 and VEGF expression were significantly greater in the Antoni B area than in the Antoni A area. Upregulated MMP-14 may degrade loose collagen in the Antoni B area and contribute to cystic formation. MMP-14 can enhance VEGF activity, which may induce extravasation of a plasma ultrafiltrate, cystic expansion, and intratumoral hemorrhage. Therefore, MMP-14 inhibition may be a therapeutic strategy for treating cystic VSs.
Matrix metalloproteinase: An upcoming therapeutic approach for idiopathic pulmonary fibrosis
2020, Pharmacological Research
Idiopathic pulmonary fibrosis (IPF) is a debilitating condition where excess collagen deposition occurs in the extracellular matrix. At first sight, it is expected that the level of different kinds of matrix metalloproteinases might be downregulated in IPF as it is a matrix degrading collagenase. However, the role of some matrix metalloproteinases (MMPs) is profibrotic where others have anti-fibrotic functions. These profibrotic MMPs effectively promote fibrosis development by stimulating the process of epithelial to mesenchymal transition. These profibrotic groups also induce macrophage polarization and fibrocyte migration. All of these events ultimately disrupt the balance between profibrotic and antifibrotic mediators, resulting aberrant repair process. Therefore, inhibition of these matrix metalloproteinases functions in IPF is a potential therapeutic approach. In addition to the use of synthetic inhibitor, various natural compounds, gene silencing act as potential natural MMP inhibitor to recover IPF.

View all citing articles on Scopus

View full text

ReviewHuman endothelial gelatinases and angiogenesis

Abstract

Introduction

Section snippets

Matrix metalloproteinases (MMPs)

Gelatinase are vital during angiogenesis

Human gelatinase B

Regulation of gelatinase B in endothelial cells

Human gelatinase A

Regulation of gelatinase A activation in endothelial cells

Current model of the role of gelatinases in angiogenesis

Conclusions and future considerations

Pharm. Therapeu.

Neurosci. Lett.

J. Biol. Chem.

Cell

Dev. Biol.

Methods Enzymol.

J. Biol. Chem.

Cell

Cell

Cell

J. Invest. Dermatol.

Am. J. Path.

J. Biol. Chem.

J. Biol. Chem.

J. Invest. Dermatol.

Inter. J. Biochem. Cell Biol.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

Inter. J. Dev. Neuro.

Angiogenesis in cancer, vascular, rheumatoid and other disease

Nature Med.

Tumour angiogenesis: therapeutic implications

New Eng. J. Med.

Basement membrane synthesis and degradation

J. Pathol.

Role of matrix metalloproteinases in human periodontal diseases

J. Periodon.

Inhibition of ovulation in the perfused rat ovary by the synthetic collagenase inhibitor SC 44463

Biol. Reprod.

Mechanism of control of trophoblast invasion in situ

J. Cell. Phys.

Expression of interstitial collagenase during skeletal development of the mouse is restricted to osteoblast-like cells and hypertrophic chondrocytes

Cell Growth Differ.

Two distinct phases of apoptosis in mammary gland involution: proteinase-independent and -dependent pathways

Development

Matrix metalloproteinases in angiogenesis: a moving target for therapeutic intervention

J. Clin. Invest.

Matrix metalloproteinases and metastasis

Cancer Chemo. Pharmacol.

Matrix metalloproteinases and their inhibitors in connective tissue remodelling

FASEB J.

The matrix-degrading metalloproteinases

Bioessays

Matrix metalloproteinase expressed on the surface of invasive tumour cells

Nature

Review
Human endothelial gelatinases and angiogenesis