Involvement of human PECAM-1 in angiogenesis and in vitro endothelial cell migration

2002 ◽  
Vol 282 (5) ◽  
pp. C1181-C1190 ◽  
Author(s):  
Gaoyuan Cao ◽  
Christopher D. O'Brien ◽  
Zhao Zhou ◽  
Samuel M. Sanders ◽  
Jordan N. Greenbaum ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Tumor Angiogenesis ◽  
Umbilical Vein ◽  
Severe Combined Immunodeficiency ◽  
Human Tumor ◽  
Tube Formation ◽  
Endothelial Cell Migration ◽  
Simultaneous Treatment

Platelet endothelial cell adhesion molecule (PECAM)-1 has been implicated in angiogenesis, but a number of issues remain unsettled, including the independent involvement of human PECAM-1 (huPECAM-1) in tumor angiogenesis and the mechanisms of its participation in vessel formation. We report for tumors grown in human skin transplanted on severe combined immunodeficiency mice that antibodies against huPECAM-1 (without simultaneous treatment with anti-VE-cadherin antibody) decreased the density of human, but not murine, vessels associated with the tumors. Anti-huPECAM-1 antibody also inhibited tube formation by human umbilical vein endothelial cells (HUVEC) and the migration of HUVEC through Matrigel-coated filters or during the repair of wounded cell monolayers. The involvement of huPECAM-1 in these processes was confirmed by the finding that expression of huPECAM-1 in cellular transfectants induced tube formation and enhanced cell motility. These data provide evidence of a role for PECAM-1 in human tumor angiogenesis (independent of VE-cadherin) and suggest that during angiogenesis PECAM-1 participates in adhesive and/or signaling phenomena required for the motility of endothelial cells and/or their subsequent organization into vascular tubes.

CYLD regulates angiogenesis by mediating vascular endothelial cell migration

Blood ◽  
2010 ◽  
Vol 115 (20) ◽  
pp. 4130-4137 ◽  
Author(s):  
Jinmin Gao ◽  
Lei Sun ◽  
Lihong Huo ◽  
Min Liu ◽  
Dengwen Li ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Migration ◽  
Endothelial Cell ◽  
Vascular Endothelial Cells ◽  
Vascular Endothelial Cell ◽  
Tube Formation ◽  
Endothelial Cell Migration ◽  
Vascular Endothelial

Cylindromatosis (CYLD) is a deubiquitinase that was initially identified as a tumor suppressor and has recently been implicated in diverse normal physiologic processes. In this study, we have investigated the involvement of CYLD in angiogenesis, the formation of new blood vessels from preexisting ones. We find that knockdown of CYLD expression significantly impairs angiogenesis in vitro in both matrigel-based tube formation assay and collagen-based 3-dimensional capillary sprouting assay. Disruption of CYLD also remarkably inhibits angiogenic response in vivo, as evidenced by diminished blood vessel growth into the angioreactors implanted in mice. Mechanistic studies show that CYLD regulates angiogenesis by mediating the spreading and migration of vascular endothelial cells. Silencing of CYLD dramatically decreases microtubule dynamics in endothelial cells and inhibits endothelial cell migration by blocking the polarization process. Furthermore, we identify Rac1 activation as an important factor contributing to the action of CYLD in regulating endothelial cell migration and angiogenesis. Our findings thus uncover a previously unrecognized role for CYLD in the angiogenic process and provide a novel mechanism for Rac1 activation during endothelial cell migration and angiogenesis.

scholarly journals Bone morphogenic protein antagonist Drm/gremlin is a novel proangiogenic factor

Blood ◽  
2006 ◽  
Vol 109 (5) ◽  
pp. 1834-1840 ◽  
Author(s):  
Helena Stabile ◽  
Stefania Mitola ◽  
Emanuela Moroni ◽  
Mirella Belleri ◽  
Stefania Nicoli ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Intracellular Signaling ◽  
Lung Tumor ◽  
Human Tumor ◽  
Bone Morphogenic Protein ◽  
Endothelial Cell Migration ◽  
Migration And Invasion ◽  
Collagen Gels

Abstract Angiogenesis plays a key role in various physiologic and pathologic conditions, including tumor growth. Drm/gremlin, a member the Dan family of bone morphogenic protein (BMP) antagonists, is commonly thought to affect different processes during growth, differentiation, and development by heterodimerizing various BMPs. Here, we identify Drm/gremlin as a novel proangiogenic factor expressed by endothelium. Indeed, Drm/gremlin was purified to homogeneity from the conditioned medium of transformed endothelial cells using an endothelial-cell sprouting assay to follow protein isolation. Accordingly, recombinant Drm/gremlin stimulates endothelial-cell migration and invasion in fibrin and collagen gels, binds with high affinity to various endothelial cell types, and triggers tyrosine phosphorylation of intracellular signaling proteins. Also, Drm/gremlin induces neovascularization in the chick embryo chorioallantoic membrane. BMP4 does not affect Drm/gremlin interaction with endothelium, and both molecules exert a proangiogenic activity in vitro and in vivo when administered alone or in combination. Finally, Drm/gremlin is produced by the stroma of human tumor xenografts in nude mice, and it is highly expressed in endothelial cells of human lung tumor vasculature when compared with non-neoplastic lung. Our observations point to a novel, previously unrecognized capacity of Drm/gremlin to interact directly with target endothelial cells and to modulate angiogenesis.

Abstract 1461: BLADE Is A Novel Membrane Protein That Regulates Endothelial Apoptosis and Consequently Angiogenesis Both In Vitro and In Vivo

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Koji Ikeda ◽  
Ritsuko Nakano ◽  
Thomas Quertermous ◽  
Hiroaki Matsubara
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Membrane Protein ◽  
Umbilical Vein ◽  
Hepatoma Cell ◽  
Endothelial Cell Migration ◽  
Late Time ◽  
Endothelial Apoptosis

Endothelial apoptosis is a pivotal process for angiogenesis during embryogenesis as well as postnatal. Here, we identified a novel gene, termed BLADE, that regulates endothelial apoptosis and consequently angiogenesis. BLADE was highly expressed in a variety of human cultured endothelial cells such as endothelial cells from umbilical vein (HUVEC), aorta, coronary artery and microvessels while no expression was observed in non-endothelial cells including HeLa cell, hepatoma cell and vascular smooth muscle cells. Furthermore, BLADE was expressed predominantly in blood vessels during embryogenesis, suggesting its role in angiogenesis. Amino acid sequence analysis revealed that BLADE was a novel membrane protein with no conserved domain reported before. BLADE expression was altered by treatment with VEGF, TNF-alpha and TGF-beta in HUVEC. Knockdown of BLADE in HUVEC using short interference RNA resulted in significant reduction of endothelial apoptosis. In contrast, BLADE knockdown affected neither endothelial cell migration nor proliferation. Among factors associated with apoptosis, Bcl-2 expression was drastically increased in HUVEC by BLADE-knockdown. Further analysis revealed that this increase of Bcl-2 is due to reduced ubiquitination and less subsequent degradation of Bcl-2. Inhibition of Bcl-2 completely abolished the anti-apoptotic effect of BLADE-knockdown. In vitro tube-formation of HUVEC on Matrigel was reduced concomitantly with reduced apoptosis by BLADE-knockdown at early time (12h). Nevertheless, significantly more tubes were preserved at late time (96h) by BLADE-knockdown. Moreover, in vivo angiogenesis assessed by Matrigel-plug was dramatically enhanced by BLADE-knockdown. Taken together, BLADE is a novel factor regulating endothelial apoptosis as well as angiogenesis both in vitro and in vivo. Detailed analysis of BLADE will provide new insights in the regulation of endothelial apoptosis, and in the molecular link between endothelial apoptosis and angiogenesis. Because BLADE is highly preferentially expressed in endothelial cells, inhibition of BLADE might be an ideal approach to enhance endothelial cell survival and angiogenesis without affecting the survival of other type of cells.

Functionally defining the endothelial transcriptome, from Robo4 to ECSCR

2009 ◽  
Vol 37 (6) ◽  
pp. 1214-1217 ◽  
Author(s):  
Ana Raquel Verissimo ◽  
John M.J. Herbert ◽  
Victoria L. Heath ◽  
John A. Legg ◽  
Helen Sheldon ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Migration ◽  
Endothelial Cell ◽  
Critical Role ◽  
Tube Formation ◽  
Endothelial Cell Migration ◽  
In Vitro Assays ◽  
Filamin A ◽  
Interfering Rna

We have applied search algorithms to expression databases to identify genes whose expression is restricted to the endothelial cell. Such genes frequently play a critical role in endothelial biology and angiogenesis. Two such genes are the roundabout receptor Robo4 and the ECSCR (endothelial-cell-specific chemotaxis regulator). Endothelial cells express both Robo1 and Robo4, which we have knocked down using siRNA (small interfering RNA) and then studied the effect in a variety of in vitro assays. Both Robo4 and Robo1 knockdown inhibited in vitro tube formation on Matrigel™. Transfection of Robo4 into endothelial cells increased the number of filopodial extensions from the cell, but failed to do so in Robo1-knockdown cells. Separate immunoprecipitation studies showed that Robo1 and Robo4 heterodimerize. We conclude from this and other work that a heteroduplex of Robo1 and Robo4 signals through WASP (Wiskott–Aldrich syndrome protein) and other actin nucleation-promoting factors to increase the number of filopodia and cell migration. Knockdown of the transmembrane ECSCR protein in endothelial cells also reduced chemotaxis and impaired tube formation on Matrigel™. Yeast two-hybrid analysis and immunoprecipitation studies showed that, in contrast with the roundabouts, ECSCR binds to the actin-modulatory filamin A. We conclude that all three of these genes are critical for effective endothelial cell migration and, in turn, angiogenesis.

scholarly journals Exosomal CircHIPK3 Released from Hypoxia-Induced Cardiomyocytes Regulates Cardiac Angiogenesis after Myocardial Infarction

2020 ◽  
Vol 2020 ◽  
pp. 1-19 ◽  
Author(s):  
Yan Wang ◽  
Ranzun Zhao ◽  
Changyin Shen ◽  
Weiwei Liu ◽  
Jinson Yuan ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Cycle Progression ◽  
Cell Activity ◽  
Cell Communication ◽  
Tube Formation ◽  
Endothelial Cell Migration ◽  
Endothelial Cell Proliferation

Exosomes play critical roles in mediating cell-to-cell communication by delivering noncoding RNAs (including miRNAs, lncRNAs, and circRNAs). Our previous study found that cardiomyocytes (CMs) subjected to hypoxia released circHIPK3-rich exosomes to regulate oxidative stress damage in cardiac endothelial cells. However, the role of exosomes in regulating angiogenesis after myocardial infarction (MI) remains unknown. The aim of this study was to establish the effects of exosomes derived from hypoxia-induced CMs on the migration and angiogenic tube formation of cardiac endothelial cells. Here, we reported that hypoxic exosomes (HPC-exos) can effectively reduce the infarct area and promote angiogenesis in the border surrounding the infarcted area. HPC-exos can also promote cardiac endothelial cell migration, proliferation, and tube formation in vitro. However, these effects were weakened after silencing circHIPK3 in hypoxia-induced CMs. We further verified that silencing and overexpressing circHIPK3 changed cardiac endothelial cell proliferation, migration, and tube formation in vitro by regulating the miR-29a expression. In addition, exosomal circHIPK3 derived from hypoxia-induced CMs first led to increased VEGFA expression by inhibiting miR-29a activity and then promoted accelerated cell cycle progression and proliferation in cardiac endothelial cells. Overexpression of miR-29a mimicked the effect of silencing circHIPK3 on cardiac endothelial cell activity in vitro. Thus, our study provides a novel mechanism by which exosomal circRNAs are involved in the communication between CMs and cardiac endothelial cells.

scholarly journals Organizational behavior of human umbilical vein endothelial cells.

1982 ◽  
Vol 94 (3) ◽  
pp. 511-520 ◽  
Author(s):  
T Maciag ◽  
J Kadish ◽  
L Wilkins ◽  
M B Stemerman ◽  
R Weinstein
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Organizational Behavior ◽  
Umbilical Vein ◽  
Ultrastructural Level ◽  
Tube Formation ◽  
Culture Conditions ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells

Culture conditions that favor rapid multiplication of human umbilical vein endothelial cells (HUV-EC) also support long-term serial propagation of the cells. This is routinely achieved when HUV-EC are grown in Medium 199 (M-199) supplemented with fetal bovine serum (FBS) and endothelial cell growth factor (ECGF), on a human fibronectin (HFN) matrix. The HUV-EC can shift from a proliferative to an organized state when the in vitro conditions are changed from those favoring low density proliferation to those supporting high density survival. When ECGF and HFN are omitted, cultures fail to achieve confluence beyond the first or second passage: the preconfluent cultures organize into tubular structures after 4-6 wk. Some tubes become grossly visible and float in the culture medium, remaining tethered to the plastic dish at either end of the tube. On an ultrastructural level, the tubes consist of cells, held together by junctional complexes, arranged so as to form a lumen. The smallest lumens are formed by one cell folding over to form a junction with itself. The cells contain Weibel-Palade bodies and factor VIII-related antigen. The lumens contain granular, fibrillar and amorphous debris. Predigesting the HFN matrix with trypsin (10 min, 37 degrees C) or plasmin significantly accelerates tube formation. Thrombin and plasminogen activator had no apparent effect. Disruption of the largest tubes with trypsin/EDTA permits the cells to revert to a proliferative state if plated on HFN, in M-199, FBS, and ECGF. These observations indicate that culture conditions that do not favor proliferation permit attainment of a state of nonterminal differentiation (organization) by the endothelial cell. Furthermore, proteolytic modification of the HFN matrix may play an important role in endothelial organization.

scholarly journals Comparative analysis of the effects of opioids in angiogenesis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Tao Feng ◽  
Si Zeng ◽  
Jie Ding ◽  
Gong Chen ◽  
Bin Wang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Umbilical Vein ◽  
Tube Formation ◽  
Mitogen Activated Protein Kinase ◽  
Mitogen Activated Protein ◽  
Apoptotic Effect ◽  
Toxicity Of Drugs ◽  
Umbilical Vein Endothelial Cells

Abstract Background Angiogenesis, the formation of blood vessel from pre-existing ones, plays an important role in many pathophysiological diseases, such as cancer. Opioids are often used in clinic for the management of chronic pain in cancer patients at terminal phases. Here, we investigated and compared the effects and mechanisms of four opioids on angiogenesis. Methods We performed angiogenesis assays on human umbilical vein endothelial cells (HUVEC) that represent an in vitro model to assess the toxicity of drugs to endothelium. Results Morphine and oxycodone at 0.1 μM to 100 μM dose-dependently increased endothelial cell tube formation and proliferation. We observed the same in endothelial cells exposed to fentanyl at 0.1 μM to 10 μM but there was a gradual loss of stimulation by fentanyl at 100 μM and 1000 μM. Morphine and fentanyl reduced endothelial cell apoptosis-induced by serum withdrawal whereas oxycodone did not display anti-apoptotic effect, via decreasing Bax level. Oxycodone at the same concentrations was less potent than morphine and fentanyl. Different from other three opioids, codeine at all tested concentrations did not affect endothelial cell tube formation, proliferation and survival. Mechanism studies demonstrated that opioids acted on endothelial cells via μ-opioid receptor-independent pathway. Although we observed the increased phosphorylation of mitogen-activated protein kinase (MAPK) in cells exposed to morphine, fentanyl and oxycodone, the rescue studies demonstrated that the stimulatory effects of morphine but not fentanyl nor oxycodone were reversed by a specific MAPK inhibitor. Conclusion Our work demonstrates the differential effects and mechanisms of opioids on angiogenesis.

scholarly journals Nogo-B receptor is essential for angiogenesis in zebrafish via Akt pathway

Blood ◽  
2010 ◽  
Vol 116 (24) ◽  
pp. 5423-5433 ◽  
Author(s):  
Baofeng Zhao ◽  
Changzoon Chun ◽  
Zhong Liu ◽  
Mark A. Horswill ◽  
Kallal Pramanik ◽  
...  
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Growth Factor ◽  
Umbilical Vein ◽  
Endothelial Cell Migration ◽  
Vascular Endothelial ◽  
Endothelial Growth Factor

Abstract Our previous work has shown that axon guidance gene family Nogo-B and its receptor (NgBR) are essential for chemotaxis and morphogenesis of endothelial cells in vitro. To investigate NogoB-NgBR function in vivo, we cloned the zebrafish ortholog of both genes and studied loss of function in vivo using morpholino antisense technology. Zebrafish ortholog of Nogo-B is expressed in somite while expression of zebrafish NgBR is localized in intersomitic vessel (ISV) and axial dorsal aorta during embryonic development. NgBR or Nogo-B knockdown embryos show defects in ISV sprouting in the zebrafish trunk. Mechanistically, we found that NgBR knockdown not only abolished its ligand Nogo-B–stimulated endothelial cell migration but also reduced the vascular endothelial growth factor (VEGF)–stimulated phosphorylation of Akt and vascular endothelial growth factor–induced chemotaxis and morphogenesis of human umbilical vein endothelial cells. Further, constitutively activated Akt (myristoylated [myr]Akt) or human NgBR can rescue the NgBR knockdown umbilical vein endothelial cell migration defects in vitro or NgBR morpholino-caused ISV defects in vivo. These data place Akt at the downstream of NgBR in both Nogo-B– and VEGF-coordinated sprouting of ISVs. In summary, this study identifies the in vivo functional role for Nogo-B and its receptor (NgBR) in angiogenesis in zebrafish.

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