Nobiletin, a citrus polymethoxyflavonoid, suppresses multiple angiogenesis-related endothelial cell functions and angiogenesis in vivo

ABSTRACTAngiogenesis is a complex morphogenetic process that involves intimate interactions between multicellular endothelial structures and their extracellular milieu. In vitro models of angiogenesis can aid in reducing the complexity of the in vivo microenvironment and provide mechanistic insight into how soluble and physical extracellular matrix cues regulate this process. To investigate how microenvironmental cues regulate angiogenesis and the function of resulting microvasculature, we multiplexed an established angiogenesis-on-a-chip platform that affords higher throughput investigation of 3D endothelial cell sprouting emanating from a parent vessel through defined biochemical gradients and extracellular matrix. We found that two fundamental endothelial cell functions, migration and proliferation, dictate endothelial cell invasion as single cells vs. multicellular sprouts. Microenvironmental cues that elicit excessive migration speed incommensurate with proliferation resulted in microvasculature with poor barrier function and an inability to transport fluid across the microvascular bed. Restoring the balance between migration speed and proliferation rate rescued multicellular sprout invasion, providing a new framework for the design of pro-angiogenic biomaterials that guide functional microvasculature formation for regenerative therapies.

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BNO 1095, a Standardized Dry Extract from the Fruits of Vitex agnus-castus, Impairs Angiogenesis-related Endothelial Cell Functions In Vitro

Planta Medica ◽

10.1055/a-1351-1038 ◽

2021 ◽

Author(s):

Iris Bischoff-Kont ◽

Laura Brabenec ◽

Rebecca Ingelfinger ◽

Bernhard Nausch ◽

Robert Fürst

Keyword(s):

Endothelial Cell ◽

Ex Vivo ◽

Collagen Gel ◽

Tissue Growth ◽

Endothelial Cell Proliferation ◽

Dry Extract ◽

Cell Functions ◽

Agnus Castus

AbstractBNO 1095, a standardized dry extract from the fruits of Vitex agnus-castus, represents an approved herbal medicinal product for the treatment of premenstrual syndrome. Angiogenesis, the formation of new blood vessels from pre-existing capillaries, plays a major role in physiological situations, such as wound healing or tissue growth in female reproductive organs, but it is also of great importance in pathophysiological conditions such as chronic inflammatory diseases or cancer. Angiogenesis is a highly regulated multi-step process consisting of distinct key events that can be influenced pharmacologically. Few studies suggested anti-angiogenic actions of V. agnus-castus fruit extracts in in vivo and ex vivo models. Here, we provide for the first time profound in vitro data on BNO 1095-derived anti-angiogenic effects focusing on distinct angiogenesis-related endothelial cell functions that are inevitable for the process of new blood vessel formation. We found that V. agnus-castus extract significantly attenuated undirected and chemotactic migration of primary human endothelial cells. Moreover, the extract efficiently inhibited endothelial cell proliferation and reduced the formation of tube-like structures on Matrigel. Of note, the treatment of endothelial cell spheroids almost blocked endothelial sprouting in a 3D collagen gel. Our data present new and detailed insights into the anti-angiogenic actions of BNO 1095 and, therefore, suggest a novel scope of potential therapeutic applications of the extract for which these anti-angiogenic properties are required.

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Extracellular Nanovesicles Secreted by Human Osteosarcoma Cells Promote Angiogenesis

Cancers ◽

10.3390/cancers11060779 ◽

2019 ◽

Vol 11 (6) ◽

pp. 779 ◽

Cited By ~ 5

Author(s):

Francesca Perut ◽

Laura Roncuzzi ◽

Nicoletta Zini ◽

Annamaria Massa ◽

Nicola Baldini

Keyword(s):

Endothelial Cell ◽

Chorioallantoic Membrane ◽

Tube Formation ◽

Osteosarcoma Cells ◽

Cell Functions ◽

Human Osteosarcoma ◽

Human Osteosarcoma Cells ◽

Neutral Conditions

Angiogenesis involves a number of different players among which extracellular nanovesicles (EVs) have recently been proposed as an efficient cargo of pro-angiogenic mediators. Angiogenesis plays a key role in osteosarcoma (OS) development and progression. Acidity is a hallmark of malignancy in a variety of cancers, including sarcomas, as a result of an increased energetic metabolism. The aim of this study was to investigate the role of EVs derived from osteosarcoma cells on angiogenesis and whether extracellular acidity, generated by tumor metabolism, could influence EVs activity. For this purpose, we purified and characterized EVs from OS cells maintained at either acidic or neutral pH. The ability of EVs to induce angiogenesis was assessed in vitro by endothelial cell tube formation and in vivo using chicken chorioallantoic membrane. Our findings demonstrated that EVs derived from osteosarcoma cells maintained either in acidic or neutral conditions induced angiogenesis. The results showed that miRNA and protein content of EVs cargo are correlated with pro-angiogenic activity and this activity is increased by the acidity of tumor microenvironment. This study provides evidence that EVs released by human osteosarcoma cells act as carriers of active angiogenic stimuli that are able to promote endothelial cell functions relevant to angiogenesis.

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New in vivo model to assess venous endothelial cell functions. Effect of defibrotide

European Journal of Clinical Pharmacology ◽

10.1007/bf00558499 ◽

1989 ◽

Vol 37 (4) ◽

pp. 351-357 ◽

Cited By ~ 4

Author(s):

T. Di Perri ◽

F. Laghi Pasini ◽

C. Frigerio ◽

P. L. Capecchi ◽

G. L. Messa ◽

...

Keyword(s):

Endothelial Cell ◽

In Vivo Model ◽

Cell Functions

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Dual role for TWEAK in angiogenic regulation

Journal of Cell Science ◽

10.1242/jcs.115.2.267 ◽

2002 ◽

Vol 115 (2) ◽

pp. 267-274 ◽

Cited By ~ 1

Author(s):

Aniela Jakubowski ◽

Beth Browning ◽

Matvey Lukashev ◽

Irene Sizing ◽

Jeffrey S. Thompson ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Growth Factor ◽

Cell Types ◽

Dual Role ◽

Epithelial Tumor ◽

Potent Inducer ◽

Angiogenic Potential ◽

Cell Functions

Angiogenic regulators modulate endothelial cell functions, including proliferation, migration, secretion, and adhesion, through their action on endothelial cells or other cell types. TWEAK, a novel member of the tumor necrosis factor family, appears to be a pro-angiogenic agent on the basis of previous studies demonstrating its ability to induce interleukin-8 production by epithelial tumor lines, stimulate proliferation of human vascular cell types and neovascularization in rat corneas. Here, we further characterized the angiogenic potential of TWEAK, revealing a dual role for TWEAK as an angiogenic regulator. We demonstrate that TWEAK is a potent inducer of endothelial cell survival and cooperates with basic fibroblast growth factor to induce the proliferation and migration of human endothelial cells and morphogenesis of capillary lumens. In contrast, TWEAK antagonizes the morphogenic response of endothelial cells to vascular endothelial growth factor (VEGF) without inhibiting VEGF-induced survival or proliferation. Thus, our observations suggest that TWEAK may differentially regulate microvascular growth, remodeling and/or maintenance in vivo, depending upon the angiogenic context.

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Ets1 and Ets2 are required for endothelial cell survival during embryonic angiogenesis

Blood ◽

10.1182/blood-2009-03-211391 ◽

2009 ◽

Vol 114 (5) ◽

pp. 1123-1130 ◽

Cited By ~ 98

Author(s):

Guo Wei ◽

Ruchika Srinivasan ◽

Carmen Z. Cantemir-Stone ◽

Sudarshana M. Sharma ◽

Ramasamy Santhanam ◽

...

Keyword(s):

Endothelial Cell ◽

Target Genes ◽

Cell Function ◽

Endothelial Cell Function ◽

Cell Functions ◽

And Function ◽

Target Promoters ◽

Vascular Branching

Abstract The ras/Raf/Mek/Erk pathway plays a central role in coordinating endothelial cell activities during angiogenesis. Transcription factors Ets1 and Ets2 are targets of ras/Erk signaling pathways that have been implicated in endothelial cell function in vitro, but their precise role in vascular formation and function in vivo remains ill-defined. In this work, mutation of both Ets1 and Ets2 resulted in embryonic lethality at midgestation, with striking defects in vascular branching having been observed. The action of these factors was endothelial cell autonomous as demonstrated using Cre/loxP technology. Analysis of Ets1/Ets2 target genes in isolated embryonic endothelial cells demonstrated down-regulation of Mmp9, Bcl-XL, and cIAP2 in double mutants versus controls, and chromatin immunoprecipitation revealed that both Ets1 and Ets2 were loaded at target promoters. Consistent with these observations, endothelial cell apoptosis was significantly increased both in vivo and in vitro when both Ets1 and Ets2 were mutated. These results establish essential and overlapping functions for Ets1 and Ets2 in coordinating endothelial cell functions with survival during embryonic angiogenesis.

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Divergent effects of quercetin conjugates on angiogenesis

British Journal Of Nutrition ◽

10.1079/bjn20061753 ◽

2006 ◽

Vol 95 (5) ◽

pp. 1016-1023 ◽

Cited By ~ 49

Author(s):

Sandra Donnini ◽

Federica Finetti ◽

Lorenzo Lusini ◽

Lucia Morbidelli ◽

Veronique Cheynier ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Inhibitory Effect ◽

Endothelial Cell Proliferation ◽

Vegf Receptor ◽

Cell Functions ◽

Extracellular Signal ◽

Opposing Effects ◽

Endothelial Growth Factor

The present study reports the activities of quercetin and its main circulating conjugates in man (quercetin-3′-sulphate (Q3′S) and quercetin-3-glucuronide (Q3G)) on in vivo angiogenesis induced by vascular endothelial growth factor (VEGF) and examines the effects of these molecules on cultured endothelial cells. We found opposing effects of quercetin and its metabolites on angiogenesis. While quercetin and Q3G inhibited VEGF-induced endothelial cell functions and angiogenesis, Q3′S per se promoted endothelial cell proliferation and angiogenesis. The inhibitory effect elicited by Q3G was linked to inhibition of extracellular signal-regulated kinases 1/2 (ERK1/2) phosphorylation elicited by VEGF. The activation of endothelial cells by Q3′S was associated to stimulation of VEGF receptor-2 and to downstream signalling activation (phosphatidylinositol-3 kinase/Akt and nitric oxide synthase pathways), ultimately responsible for ERK1/2 phosphorylation. These data indicate that the effects of circulating quercetin conjugates on angiogenesis are different depending on the nature of the conjugate. Q3G andQ3′S are the two major conjugates in plasma, but their ratio is dependenton several factors, so thatinhibition or activation of angiogenesis could be subtly shifted as a result of metabolismin vivo.

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Induction and detection of a human endothelial activation antigen in vivo.

Journal of Experimental Medicine ◽

10.1084/jem.164.2.661 ◽

1986 ◽

Vol 164 (2) ◽

pp. 661-666 ◽

Cited By ~ 243

Author(s):

R S Cotran ◽

M A Gimbrone ◽

M P Bevilacqua ◽

D L Mendrick ◽

J S Pober

Keyword(s):

Endothelial Cell ◽

Umbilical Vein ◽

Endothelial Activation ◽

Immunoperoxidase Technique ◽

Positive Staining ◽

Inflammatory Reactions ◽

Cell Functions ◽

Activation Antigen

We used a murine mAb, H4/18, raised by immunization with IL-1-treated human umbilical vein endothelial cell cultures, to localize an endothelial activation antigen in induced human delayed hypersensitivity reactions (DHR) and in pathological tissues. We used streptococcus varidase to elicit DHR in human skin and we examined sequential skin biopsies with the immunoperoxidase technique. There was no staining for H4/18 binding antigen in normal endothelium of skin and other tissues; strong positive staining, localized to vascular endothelium, was seen at 16 and 23 h but disappeared by 6 d, when the DHR had faded. H4/18 binding antigen, also confined to endothelium, was detected in lymph nodes, skin, and other tissues exhibiting immune/inflammatory reactions. The studies indicate that H4/18 is a useful marker for activated endothelium in vivo and they support the relevance of in vitro studies on inducible endothelial cell functions.

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