Input:
Evaluation of electrical stimulation for ischemic wound therapy: a feasibility study using the lapine wound model.

Chronic wounds are a major secondary complication for many people with impaired mobility. Electrical stimulation (ES) has been recommended as a adjunctive therapy, however optimal treatment paradigms have not been established. Our group seeks to determine the basic mechanisms underlying ES wound therapy, an area where understanding is currently limited. A feasibility study was carried out to develop the Ahn/Mustoe lapine wound model for systematic investigation of the effects of electrical stimulation on ischemic wound therapy. A standardized surgical procedure incorporated a hybrid stimulation system comprising an implantable mini-stimulator and surface electrodes, with creation of repeatable ischemic wounds. Twenty mature male New Zealand white rabbits (3 kg weight) were employed to evaluate the effects of two empirically selected stimulation paradigms applied continuously for 7-21 days, using each animal as its own control. Outcomes measures included transcutaneous blood gas levels, histology, total RNA content and analysis of alpha2 (I) collagen (COL-I), type IV collagen (COL-IV), alpha1 (V) collagen (COL-V), and vascular endothelial growth factor (VEGF) expression using real-time quantitative PCR. All markers for stimulated wounds showed increased activity relative to non-stimulated control wounds between 7 and 14 days following injury, with peak activity at 14 days. By 21 days post-injury, all activity had returned to near baseline level. VEGF and COL-IV levels were found to be significantly higher for pattern A (110 mus pulse width) compared to pattern B (5 mus pulse width) at 14 days, implying that pattern A may be more effective at promoting angiogenesis. All wounds were fully re-epithelialized by 10 days post-injury. Both COL-I and COL-V showed statistically significant (P  less than  0.05) increased activity between day 7 and day 14 for pattern A, potentially indicating a continued effect on matrix remodeling. The early closure of all wounds implies that the rabbit ear model may not be valid for chronic wound studies.

angiogenesis

expression

alpha2 (I) collagen

expression

type IV collagen

expression

alpha1 (V) collagen

expression

vascular endothelial growth factor

promoting

angiogenesis

activity

COL-I

activity

COL-V

effect

COL-I

remodeling

effect

COL-V

remodeling

remodeling

matrix


Input:
VEGF, angiopoietin-1 and -2 in bronchial asthma: new molecular targets in airway angiogenesis and microvascular remodeling.

Airway angiogenesis and microvascular remodeling are known features of bronchial asthma, but the mechanisms of these structural alterations are just beginning to be elucidated. Vascular endothelial growth factor (VEGF), one of the most potent angiogenic factors, stimulates endothelial cell proliferation and induces the angiogenesis. Recently, considerable attentions have been devoted to the physiological roles of angiopoietin (Ang)-1 and -2 as regulatory factors of VEGF. Ang-1 has been shown to induce the migration and sprouting of endothelial cells, and coexpression of Ang-1 and VEGF enhanced angiogenesis. In the presence of high levels of VEGF, Ang-2 also promotes rapid increase in capillary diameter, remodeling of the basal lamina, proliferation and migration of endothelial cells, and stimulates sprouting of new blood vessels. Thus, VEGF, Ang-1 and -2 may play complementary and coordinated roles in airway angiogenesis and microvascular remodeling, and these structural changes are potentially reversible by therapeutic intervention. The scope of the present review is to discuss from a clinical point of view the potential interactions between VEGF and angiopoietins in the asthmatic airways, and focus on the therapeutic implications targeting for these angiogenic factors. Recently, there is an increasing number of patents which have been focused on the inhibitors of VEGF action. These inhibitors are directed towards the receptors of VEGF or intracellular substrates for the receptors. We will also discuss several patents regarding inhibitors of VEGF action in the present review.

interactions

VEGF

angiopoietins

angiogenesis

angiogenesis

angiogenic

angiogenesis

angiogenesis

angiogenesis

angiogenic

proliferation

endothelial cell

proliferation

endothelial cells

sprouting

endothelial cells

sprouting

blood vessels

coexpression

VEGF

migration

endothelial cells

presence

VEGF

migration

endothelial cells

stimulates

Vascular endothelial growth factor

proliferation

induces

Vascular endothelial growth factor

angiogenesis

induce

migration

induce

sprouting

enhanced

coexpression

angiogenesis

promotes

capillary

promotes

remodeling

promotes

proliferation

promotes

migration

stimulates

sprouting

play complementary and coordinated roles

VEGF

angiogenesis

play complementary and coordinated roles

VEGF

remodeling

play complementary and coordinated roles

angiogenesis

play complementary and coordinated roles

remodeling

play complementary and coordinated roles

angiogenesis

play complementary and coordinated roles

remodeling

remodeling

microvascular

remodeling

microvascular

remodeling

basal lamina

remodeling

microvascular


Input:
Effect of transplanted mesenchymal stem cells from rats of different ages on the improvement of heart function after acute myocardial infarction.

BACKGROUND: Mesenchymal stem cells (MSCs) transplantation is of therapeutic potential after ischemic injury in both experimental and clinical studies. Clinically, elderly patients are more vulnerable to acute myocardial infarction (AMI). But little is known about the characteristics of young donor-derived MSCs transplanted to old patients with AMI. The present study was designed to investigate the effect of transplanted MSCs from rats of different ages on the improvement of heart function after AMI. METHODS: MSCs from Sprague-Dawley (SD) rats were isolated and cultured in vitro. The apoptosis characteristics of MSCs were observed under conditions of ischemia and anoxia. SD rats underwent MI received intramyocardial injection of MSCs from young donor rats (n = 8), old donor rats (n = 8), respectively. AMI control group received equal volume physiological saline. Immunofluorescence was used to observe the differentiation of the grafted cells into cardiomyocytes. Four weeks after cell transplantation, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry for vascular endothelial growth factor (VEGF), VIII-factor immunohistochemistry for vessel density, TUNEL, caspase-3 for cardiomyocyte apoptosis, echocardiography and hemodynamic detection for heart function were performed. RESULTS: The apoptosis rate of the old donor-derived MSCs group was significantly higher than that of the young donor-derived MSCs group under conditions of ischemia and anoxia (P  less than  0.05). Engrafted MSCs survived, proliferated and differentiated into myocardium-like cells. VEGF gene expression and capillary density in the old donor-derived group were lower than those in the young donor-derived group but higher than those in the control group (P  less than  0.05). The transplantation of old donor-derived MSCs attenuated apoptosis of cardiomyocytes in the peri-infarct region compared with the control group and the effect was elevated in young donor-derived MSCs (P  less than  0.05). The heart functions (left ventricle ejection fraction (LVEF), left ventricle fractional shortening (LVFS)) were improved more significantly in the old donor-derived MSCs group than in the control group and the heart function in the young donor-derived MSCs group further improved (P  less than  0.05). CONCLUSIONS: Young donor-derived MSCs can improve heart function significantly through angiogenesis and decreasing cardiomyocyte apoptosis when transplanted to the infarcted area.

angiogenesis

proliferated

MSCs

apoptosis

MSCs

apoptosis

cardiomyocyte

apoptosis

MSCs

apoptosis

MSCs

survived

MSCs

apoptosis

cardiomyocytes

apoptosis

cardiomyocyte

differentiation

cells

differentiated

MSCs

expression

VEGF

attenuated

transplantation

apoptosis

decreasing

MSCs

apoptosis

transplanted

mesenchymal stem cells

transplantation

Mesenchymal stem cells

transplanted

MSCs

patients

transplanted

MSCs

isolated

MSCs

Sprague-Dawley (SD) rats

cultured

MSCs

injection

SD rats

MSCs

grafted

cells

transplantation

cell

Engrafted

MSCs

transplantation

MSCs

transplanted

MSCs

elevated

effect

improved

heart

improve

improve

heart

effect

MSCs

improve

MSCs

angiogenesis


Input:
Clinical significance of chicken ovalbumin upstream promoter-transcription factor II expression in human colorectal cancer.

Chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) plays an essential role in angiogenesis and development. A previous study showed that the expression of COUP-TFII enhanced invasiveness of human lung carcinoma cells. However, no published data are available concerning the biological and clinical significance of COUP-TFII expression in colorectal cancer. Thus, our objective was to explore the expression of COUP-TFII in colorectal cancer as well as its association with clinicopathological features, and to evaluate the role of COUP-TFII as a prognostic indicator in colorectal cancer. We investigated the presence of COUP-TFII in human colorectal cancer tissues and adjacent normal tissues from 95 primary colorectal cancer patients by immunohistochemistry. The correlation between the expression of COUP-TFII and clinicopathologic features was investigated. The 3-year disease-free survival (DFS) and overall survival (OS) of patients with tumors expressing different levels of COUP-TFII were evaluated by the Kaplan-Meier method. No significant correlation was found between COUP-TFII expression and age at surgery, gender, histopathologic differentiation, vessel invasion, carcinoembryonic antigen (CEA), or nodal involvement. However, survival analysis showed that the COUP-TFII-positive group had a significantly better OS compared to COUP-TFII-negative group (80.4% vs. 57.7%, P=0.0491). Based on our results, COUP-TFII may represent a biomarker for good prognosis in colorectal cancer.

angiogenesis

expression

chicken ovalbumin upstream promoter-transcription factor II

expression

COUP-TFII

expression

COUP-TFII

expression

COUP-TFII

expression

COUP-TFII

expressing

COUP-TFII

expression

COUP-TFII

positive

COUP-TFII

negative

COUP-TFII

invasiveness

lung carcinoma cells

presence

COUP-TFII

colorectal cancer tissues

presence

COUP-TFII

normal tissues

invasion

vessel

enhanced

expression

invasiveness

plays an essential role

Chicken ovalbumin upstream promoter-transcription factor II

angiogenesis

role

COUP-TFII

colorectal cancer


Input:
Involvement of PTEN promoter methylation in cerebral cavernous malformations.

BACKGROUND AND PURPOSE: Cerebral cavernous malformations (CCMs) are prevalent cerebral vascular lesions involving aberrant angiogenesis. However, the underlying mechanism is poorly understood. Phosphatase and tension homolog deleted on chromosome 10 (PTEN), a tumor suppressor, is frequently deficient in various pathologies due to mutation or epigenetic alterations. PTEN promoter hypermethylation is a major epigenetic silencing mechanism leading to activation of angiogenesis in tumors. The present study aimed to investigate whether PTEN promoter methylation was involved in CCMs. METHODS: PTEN promoter methylation was detected in surgical specimens of CCMs (n=69) by methylation-specific polymerase chain reaction. The methylation status was correlated to the clinical manifestations and to PTEN expression, which was analyzed by both Western blot and immunohistochemistry. To investigate the endothelial proliferation and the potential signaling pathways affected by PTEN methylation, proliferating cell nuclear antigen as well as phosphor-Akt and phosphor-Erk1,2 were detected by immunofluorescence and Western blot, respectively, in CCM specimens. RESULTS: Methylation-specific polymerase chain reaction revealed PTEN promoter methylation in 15.9% CCMs. Strikingly, 5 of 6 familial CCMs showed PTEN promoter methylation (83.3%), which was significantly higher than in sporadic cases (9.4%; P<0.001). In addition, PTEN promoter methylation appeared more frequently in multiple CCMs, including familial cases (46.7%), than that in single-lesioned CCMs (11.8%; P<0.05). Immunostaining and Western blot revealed a more significant PTEN downregulation in PTEN-methylated CCMs in comparison to PTEN-unmethylated CCMs. Reduced PTEN expression was inversely correlated to the expression of proliferating cell nuclear antigen and to the activation of Erk1,2, but not of Akt. CONCLUSIONS: We reported here for the first time the involvement of PTEN promoter methylation in CCMs, particularly in familial CCMs, suggesting this epigenetic alteration as a potential pathomechanism of CCMs. The identification of Erk1,2 as triggered signaling in the lesions may be valuable for the development of effective therapy for this disease.

angiogenesis

angiogenesis

tumors

proliferation

endothelial

methylation

PTEN

promoter

hypermethylation

PTEN

promoter

methylation

PTEN

promoter

methylation

PTEN

promoter

methylation

PTEN

methylation

PTEN

promoter

methylation

PTEN

promoter

methylation

PTEN

promoter

methylated

PTEN

unmethylated

PTEN

methylation

PTEN

promoter

deficient

Phosphatase and tension homolog deleted on chromosome 10

expression

PTEN

expression

PTEN

expression

proliferating cell nuclear antigen

downregulation

PTEN

Reduced

expression

leading

hypermethylation

activation

activation

angiogenesis

activation

activation

activation

Akt

affected

proliferation

methylation


Input:
Aflibercept (AVE0005): an alternative strategy for inhibiting tumour angiogenesis by vascular endothelial growth factors.

BACKGROUND: Aberrant angiogenesis is a landmark feature in cancer, which is important for proliferation, growth and metastasis, and is mediated by various pro-angiogenic factors. The VEGF pathway is one of the most important and best-studied angiogenic pathways. Inhibition of this pathway may provide clinical benefits to cancer patients. OBJECTIVES: Strategies to inhibit the VEGF pathway, including antibodies to VEGF, antibodies to the extracellular domain of VEGFR-1 or VEGFR-2, decoy receptors for VEGF and tyrosine kinase inhibitors of VEGFRs, are summarized. METHODS: This review outlines and compares the latest development of these strategies, with emphasis on aflibercept, a novel decoy fusion protein of domain 2 of VEGFR-1 and domain 3 of VEGFR-2 with the Fc fragment of IgG1. RESULTS: Aflibercept was shown to have early clinical activity. Multiple studies are ongoing to determine the clinical benefits of aflibercept in cancer patients.

angiogenesis

tumour

angiogenesis

angiogenic

angiogenic

inhibiting

angiogenesis

vascular endothelial growth factors

Inhibition

pathway

inhibit

pathway

antibodies to VEGF

inhibit

pathway

antibodies to the extracellular domain of VEGFR-1

inhibit

pathway

inhibit

pathway

tyrosine kinase inhibitors

inhibit

pathway

pathway

VEGF

pathway

VEGF

Aberrant

angiogenesis

mediated

angiogenesis


Input:
The effect of perfluorocarbon-based artificial oxygen carriers on tissue-engineered trachea.

The biological effect of the perfluorocarbon-based artificial oxygen carrier (Oxygent) was investigated in tissue-engineered trachea (TET) construction. Media supplemented with and without 10% Oxygent were compared in all assessments. Partial tissue oxygen tension (PtO(2)) was measured with polarographic microprobes; epithelial metabolism was monitored by microdialysis inside the TET epithelium perfused with the medium underneath. Chondrocyte-DegraPol constructs were cultured for 1 month with the medium before glycosaminoglycan assessment and histology. Tissue reaction of TET epithelial scaffolds immersed with the medium was evaluated on the chick embryo chorioallantoic membrane. Oxygent perfusion medium increased the TET epithelial PtO(2) (51.2 +/- 0.3 mm Hg vs. 33.4 +/- 0.3 mm Hg at 200 microm thickness; 12.5 +/- 0.1 mm Hg vs. 3.1 +/- 0.1 mm Hg at 400 microm thickness, p  less than  0.01) and decreased the lactate concentration (0.63 +/- 0.08 vs. 0.80 +/- 0.06 mmol/L, p  less than  0.05), lactate/pyruvate (1.87 +/- 0.26 vs. 3.36 +/- 10.13, p  less than  0.05), and lactate/glucose ratios (0.10 +/- 0.00 vs. 0.29 +/- 0.14, p  less than  0.05). Chondrocyte-DegraPol in Oxygent group presented lower glycosaminoglycan value (0.03 +/- 0.00 vs. 0.13 +/- 0.00, p  less than  0.05); histology slides showed poor acid mucopolysaccharides formation. Orthogonal polarization spectral imaging showed no difference in functional capillary density between the scaffolds cultured on chorioallantoic membranes. The foreign body reaction was similar in both groups. We conclude that Oxygent increases TET epithelial PtO(2), improves epithelial metabolism, does not impair angiogenesis, and tends to slow cartilage tissue formation.

angiogenesis

formation

cartilage tissue

impair

Oxygent

angiogenesis

slow

Oxygent

formation

cultured

capillary

increases

Oxygent

TET epithelial

effect

tissue-engineered trachea

effect

Oxygent

tissue-engineered trachea

formation

mucopolysaccharides


Input:
The semaphorin 7A receptor Plexin C1 is lost during melanoma metastasis.

The transformation of normal melanocytes, or melanocyte stem cells, to melanoma, is a complex process involving multiple mechanisms. Loss of tumor suppressor proteins, which function as brakes on cell growth, migration, or cell survival, was recognized early on as an important mechanism for initiation and progression of melanoma. Semaphorins and their cognate receptors, Plexins and neuropilins, are involved in neuronal pathfinding, immune function, and tumor progression through effects on blood vessel growth and cell migration. Semaphorin 7A (Sema7A) is a membrane-linked semaphorin that is expressed by human keratinocytes, and we have shown that Sema7A binds to human melanocytes through beta1-integrins and the Plexin C1 receptor. Functional studies showed that Sema7A stimulates cytoskeletal reorganization in human melanocytes, resulting in adhesion and dendrite formation. Downstream targets of Plexin C1 signaling in human melanocytes include cofilin and LIM kinase II, both of which are critical mediators of cell adhesion and migration. In this report, we analyzed the expression of Plexin C1 using immunohistochemistry on sections of primary and matched metastatic lesions from 19 subjects and in a large melanoma tumor microarray. Our data show a significant loss of Plexin C1 in metastatic melanoma compared with primary melanoma, suggesting the possibility that the Plexin C1 receptor is a tumor suppressor protein for melanoma.

linked

Semaphorin 7A

membrane

binds

melanocytes

binds

binds

adhesion

melanocytes

adhesion

cell

growth

cell

survival

cell

transformation

melanoma

progression

tumor

dendrite formation

melanocytes

expressed

Semaphorin 7A

expression

Plexin C1

growth

blood vessel

metastasis

melanoma

migration

cell

migration

cell

migration

cell

brakes

growth

brakes

migration

brakes

survival

signaling

Plexin C1

through

progression

effects

stimulates

reorganization

resulting

adhesion

resulting

dendrite formation

effects

Semaphorins

growth

effects

Semaphorins

migration

effects

Plexins

growth

effects

neuropilins

growth

reorganization

cytoskeletal

targets

signaling

cofilin

targets

signaling

LIM kinase II


Input:
Characterization of the metabolic changes underlying growth factor angiogenic activation: identification of new potential therapeutic targets.

Angiogenesis is a fundamental process to normal and abnormal tissue growth and repair, which consists of recruiting endothelial cells toward an angiogenic stimulus. The cells subsequently proliferate and differentiate to form endothelial tubes and capillary-like structures. Little is known about the metabolic adaptation of endothelial cells through such a transformation. We studied the metabolic changes of endothelial cell activation by growth factors using human umbilical vein endothelial cells (HUVECs), [1,2-(13)C(2)]-glucose and mass isotopomer distribution analysis. The metabolism of [1,2-(13)C(2)]-glucose by HUVEC allows us to trace many of the main glucose metabolic pathways, including glycogen synthesis, the pentose cycle and the glycolytic pathways. So we established that these pathways were crucial to endothelial cell proliferation under vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) stimulation. A specific VEGF receptor-2 inhibitor demonstrated the importance of glycogen metabolism and pentose cycle pathway. Furthermore, we showed that glycogen was depleted in a low glucose medium, but conserved under hypoxic conditions. Finally, we demonstrated that direct inhibition of key enzymes to glycogen metabolism and pentose phosphate pathways reduced HUVEC viability and migration. In this regard, inhibitors of these pathways have been shown to be effective antitumoral agents. To sum up, our data suggest that the inhibition of metabolic pathways offers a novel and powerful therapeutic approach, which simultaneously inhibits tumor cell proliferation and tumor-induced angiogenesis.

angiogenic

Angiogenesis

angiogenic

angiogenesis

depleted

glycogen

proliferate

cells

proliferation

endothelial cell

proliferation

tumor cell

repair

tissue

differentiate

cells

form

endothelial tubes

form

capillary-like structures

transformation

endothelial cells

growth

tissue

recruiting

endothelial cells

migration

HUVEC

metabolism

metabolism

glycogen

metabolism

glycogen

reduced

migration

inhibits

proliferation

inhibits

angiogenesis

metabolic pathways

glucose

cycle

pentose

glycolytic pathways

cycle pathway

pentose

phosphate pathways

pentose

activation

angiogenic

activation

endothelial cell

stimulation

vascular endothelial growth factor

stimulation

fibroblast growth factor

induced

tumor

angiogenesis

changes

activation

synthesis

glycogen


Input:
Rab GTPase regulation of VEGFR2 trafficking and signaling in endothelial cells.

OBJECTIVE: Vascular endothelial growth factor receptor 2 (VEGFR2) is a receptor tyrosine kinase that regulates vascular physiology. However, mechanism(s) by which VEGFR2 signaling and trafficking is coordinated are not clear. Here, we have tested endocytic Rab GTPases for regulation of VEGFR2 trafficking and signaling linked to endothelial cell migration. METHODS AND RESULTS: Quiescent VEGFR2 displays endosomal localization and colocalization with the Rab5a GTPase, an early endosome fusion regulator. Expression of GTP or GDP-bound Rab5a mutants block activated VEGFR2 trafficking and degradation. Manipulation of Rab7a GTPase activity associated with late endosomes using overexpression of wild-type or mutant proteins blocks activated VEGFR2 trafficking and degradation. Depletion of Rab7a decreased VEGFR2 Y1175 phosphorylation but increased p42/44 (pERK1/2) MAPK phosphorylation. Endothelial cell migration was increased by Rab5a depletion but decreased by Rab7a depletion. CONCLUSIONS: Rab5a and Rab7a regulate VEGFR2 trafficking toward early and late endosomes. Our data suggest that VEGFR2-mediated regulation of endothelial function is dependent on different but specific Rab-mediated GTP hydrolysis activity required for endosomal trafficking.

bound

GTP

bound

GDP

degradation

degradation

hydrolysis

GTP

Expression

overexpression

Rab7a GTPase

trafficking

trafficking

trafficking

migration

endothelial cell

localization

colocalization

Rab5a GTPase

trafficking

trafficking

migration

Endothelial cell

trafficking

block

Expression

trafficking

block

Expression

degradation

blocks

overexpression

trafficking

blocks

overexpression

degradation

Depletion

decreased

Depletion

phosphorylation

depletion

decreased

migration

depletion

depletion

signaling

signaling

phosphorylation

phosphorylation

activated

trafficking

activated

degradation

activated

trafficking

activated

degradation

increased

Depletion

phosphorylation

increased

migration

depletion

dependent

regulation

hydrolysis

regulation

Rab GTPase

trafficking

regulates

Vascular endothelial growth factor receptor 2

vascular

regulation

Rab GTPases

trafficking

regulation

Rab GTPases

signaling

linked

regulation

migration

Manipulation

Rab7a GTPase

regulate

trafficking

regulate

trafficking

mediated

regulation

regulation

endothelial

mediated

Rab

hydrolysis


Input:
AngiomiRs--key regulators of angiogenesis.

The formation of new blood vessels through the process of angiogenesis is critical in vascular development and homeostasis. Aberrant angiogenesis leads to a variety of diseases, such as ischemia and cancer. Recent studies have revealed important roles for miRNAs in regulating endothelial cell (EC) function, especially angiogenesis. Mice with EC-specific deletion of Dicer, a key enzyme for generating miRNAs, display defective postnatal angiogenesis. Specific miRNAs (angiomiRs) have recently been shown to regulate angiogenesis in vivo. miRNA-126, an EC-restricted miRNA, regulates vascular integrity and developmental angiogenesis. miR-378, miR-296, and the miR-17-92 cluster contribute to tumor angiogenesis. Manipulating angiomiRs in the settings of pathological vascularization represents a new therapeutic approach.

angiogenesis

angiogenesis

angiogenesis

angiogenesis

angiogenesis

angiogenesis

angiogenesis

angiogenesis

tumor

vascularization

formation

blood vessels

development

vascular

deletion

Dicer

critical

formation

development

leads

angiogenesis

cancer

Aberrant

angiogenesis

regulating

endothelial cell

regulating

angiogenesis

regulate

angiomiRs

angiogenesis

regulates

vascular

regulates

angiogenesis

contribute

angiogenesis

contribute

angiogenesis

contribute

angiogenesis

contribute

angiogenesis


Input:
Promoting angiogenesis via manipulation of VEGF responsiveness with notch signaling.

Promoting angiogenesis via delivery of vascular endothelial growth factor (VEGF) and other angiogenic factors is both a potential therapy for cardiovascular diseases and a critical aspect for tissue regeneration. The recent demonstration that VEGF signaling is modulated by the Notch signaling pathway, however, suggests that inhibiting Notch signaling may enhance regional neovascularization, by altering the responsiveness of local endothelial cells to angiogenic stimuli. We tested this possibility with in vitro assays using human endothelial cells, as well as in a rodent hindlimb ischemia model. Treatment of cultured human endothelial cells with DAPT, a gamma secretase inhibitor, increased cell migration and sprout formation in response to VEGF stimulation with a biphasic dependence on DAPT concentration. Further, delivery of an appropriate combination of DAPT and VEGF from an injectable alginate hydrogel system into ischemic hindlimbs led to a faster recovery of blood flow than VEGF or DAPT alone; perfusion levels reached 80% of the normal level by week 4 with combined DAPT and VEGF delivery. Direct intramuscular or intraperitoneal injection of DAPT did not result in the same level of improvement, suggesting that appropriate presentation of DAPT (gel delivery) is important for its activity. DAPT delivery from the hydrogels also did not lead to any adverse side effects, in contrast to systemic introduction of DAPT. Altogether, these results suggest a new approach to promote angiogenesis by controlling Notch signaling, and may provide new options to treat patients with diseases that diminish angiogenic responsiveness.

angiogenesis

angiogenesis

angiogenic

neovascularization

angiogenic

angiogenesis

angiogenic

regeneration

tissue

formation

sprout

delivery

vascular endothelial growth factor

migration

cell

delivery

DAPT

delivery

VEGF

delivery

DAPT

inhibiting

Notch signaling

diminish

angiogenic

notch signaling

signaling

VEGF

Notch signaling pathway

Notch signaling

Notch signaling

Treatment

endothelial cells

DAPT

cultured

endothelial cells

injection

DAPT

treat

patients

Promoting

angiogenesis

manipulation

Promoting

angiogenesis

delivery

enhance

inhibiting

neovascularization

increased

migration

stimulation

increased

formation

stimulation

stimulation

Treatment

VEGF

introduction

DAPT

promote

angiogenesis

controlling

manipulation

VEGF

notch signaling

modulated

signaling

Notch signaling pathway

responsiveness

endothelial cells

controlling

Notch signaling


Input:
[Activation of sterol regulatory element binding protein and its involvement in endothelial cell migration]

OBJECTIVE: To study the activation of sterol regulatory element binding protein (SREBP) and its critical role in endothelial cell migration. METHODS: Bovine aortic endothelial cells (ECs) were cultured. The expression of SREBP and Cdc42 were determined by Western blot and quantitative real-time PCR. Moreover, outward growth migration model and transwell chamber assay were used to detect ECs migration. RESULTS: (1) SREBP was activated during ECs migration. Western blot analysis demonstrated increased active form SREBP in migrating as compared to non-migrating ECs population. SREBP activation decreased as ECs migration slowed;(2) Coincidental with SREBP activation, mRNA expression of its target genes such as low density lipoprotein receptor, HMG-CoA reductase, and fatty acid synthase also increased in migrating ECs population as detected by real-time PCR; (3) Migration induced SREBP activation in ECs was inhibited by SREBP-acting protein RNAi and pharmacologically by 25-hydroxycholesterol; (4) Inhibition of SREBP led to decreased ECs migration in various models; (5) Cells genetically deficient in SREBP-acting protein, S1P, or S2P, phenotypically exhibited impaired migration; (6) SREBP inhibition in ECs suppressed the activity of small GTPase Cdc42, a key molecule for ECs motility. CONCLUSIONS: SREBP is activated during and plays a critical role in ECs migration. Targeting SREBP could become a novel approach in fighting diseases involving abnormal ECs migration.

expression

SREBP

expression

deficient

SREBP-acting protein

deficient

deficient

migration

endothelial cell

migration

endothelial cell

migration

ECs

migration

ECs

migrating

ECs

non-migrating

ECs

migration

ECs

migrating

ECs

Migration

ECs

migration

ECs

motility

ECs

migration

ECs

migration

ECs

decreased

activation

slowed

migration

inhibited

Migration

SREBP-acting protein

inhibited

Migration

Inhibition

SREBP

decreased

migration

inhibition

SREBP

suppressed

inhibition

cultured

aortic endothelial cells

Targeting

SREBP

Activation

sterol regulatory element binding protein

activation

sterol regulatory element binding protein

activated

SREBP

increased

SREBP

activation

SREBP

activation

SREBP

increased

mRNA expression

induced

Migration

activation

activation

SREBP

led

Inhibition

decreased

activated

SREBP

involvement

sterol regulatory element binding protein

migration

critical role

sterol regulatory element binding protein

migration

plays a critical role

SREBP

migration

mRNA expression

low density lipoprotein receptor

mRNA expression

HMG-CoA reductase

mRNA expression

fatty acid synthase


Input:
Fibroblast growth factor receptor 2-positive fibroblasts provide a suitable microenvironment for tumor development and progression in esophageal carcinoma.

PURPOSE: Tumor fibroblasts (TF) have been suggested to play an essential role in the complex process of tumor-stroma interactions and tumorigenesis. The aim of the present study was to investigate the specific role of TF in the esophageal cancer microenvironment. EXPERIMENTAL DESIGN: An Affymetrix expression microarray was used to compare gene expression profiles between six pairs of TFs and normal fibroblasts from esophageal squamous cell carcinoma (ESCC). Differentially expressed genes were identified, and a subset was evaluated by quantitative real-time PCR and immunohistochemistry. RESULTS: About 43% (126 of 292) of known deregulated genes in TFs were associated with cell proliferation, extracellular matrix remodeling, and immune response. Up-regulation of fibroblast growth factor receptor 2 (FGFR2), which showed the most significant change, was detected in all six tested TFs compared with their paired normal fibroblasts. A further study found that FGFR2-positive fibroblasts were only observed inside the tumor tissues and not in tumor-surrounding stromal tissues, suggesting that FGFR2 could be used as a TF-specific marker in ESCC. Moreover, the conditioned medium from TFs was found to be able to promote ESCC tumor cell growth, migration, and invasion in vitro. CONCLUSIONS: Our study provides new candidate genes for the esophageal cancer microenvironment. Based on our results, we hypothesize that FGFR2(+)-TFs might provide cancer cells with a suitable microenvironment via secretion of proteins that could promote cancer development and progression through stimulation of cancer cell proliferation, induction of angiogenesis, inhibition of cell adhesion, enhancement of cell mobility, and promotion of the epithelial-mesenchymal transition.

interactions

tumor

stroma

adhesion

cell

angiogenesis

proliferation

cell

growth

ESCC tumor cell

proliferation

cancer cell

development

esophageal carcinoma

progression

esophageal carcinoma

transition

epithelial

transition

mesenchymal

migration

ESCC tumor cell

invasion

ESCC tumor cell

mobility

cell

inhibition

adhesion

Up-regulation

fibroblast growth factor receptor 2

stimulation

proliferation

induction

angiogenesis

enhancement

mobility

promotion

transition

play an essential role

Tumor fibroblasts

interactions

remodeling

extracellular matrix


Input:
[Role of the tumor suppressor ARF in oncogenesis]

The paper reviews the data available in the literature on a role of the tumor suppressor ARF in oncogenesis and considers the structure of a gene encoding ARF protein. The p53-dependent and p-53-independent functions of this protein are under many studies. There is evidence for the implication of ARF in angiogenesis. There is more and more information on the role of ARF in the regulation of a cell cycle, apoptosis, and autophagy. The importance of this tumor suppressor in the mechanisms of carcinogenesis is beyond question as the inactivation of ARF suppressor activity leads to the rapid growth of neoplasia. However, the exact mechanisms of ARF action yet remain unclear and require further studies by different specialists at both the molecular genetic and other levels of investigation.

angiogenesis

apoptosis

cell

growth

neoplasia

inactivation

ARF

dependent

ARF

leads

inactivation

growth

independent

ARF

implication

ARF

angiogenesis

regulation

apoptosis


Input:
Mechanical regulation of the proangiogenic factor CCN1/CYR61 gene requires the combined activities of MRTF-A and CREB-binding protein histone acetyltransferase.

Smooth muscle-rich tissues respond to mechanical overload by an adaptive hypertrophic growth combined with activation of angiogenesis, which potentiates their mechanical overload-bearing capabilities. Neovascularization is associated with mechanical strain-dependent induction of angiogenic factors such as CCN1, an immediate-early gene-encoded matricellular molecule critical for vascular development and repair. Here we have demonstrated that mechanical strain-dependent induction of the CCN1 gene involves signaling cascades through RhoA-mediated actin remodeling and the p38 stress-activated protein kinase (SAPK). Actin signaling controls serum response factor (SRF) activity via SRF interaction with the myocardin-related transcriptional activator (MRTF)-A and tethering to a single CArG box sequence within the CCN1 promoter. Such activity was abolished in mechanically stimulated mouse MRTF-A(-/-) cells or upon inhibition of CREB-binding protein (CBP) histone acetyltransferase (HAT) either pharmacologically or by siRNAs. Mechanical strain induced CBP-mediated acetylation of histones 3 and 4 at the SRF-binding site and within the CCN1 gene coding region. Inhibition of p38 SAPK reduced CBP HAT activity and its recruitment to the SRF.MRTF-A complex, whereas enforced induction of p38 by upstream activators (e.g. MKK3 and MKK6) enhanced both CBP HAT and CCN1 promoter activities. Similarly, mechanical overload-induced CCN1 gene expression in vivo was associated with nuclear localization of MRTF-A and enrichment of the CCN1 promoter with both MRTF-A and acetylated histone H3. Taken together, these data suggest that signal-controlled activation of SRF, MRTF-A, and CBP provides a novel connection between mechanical stimuli and angiogenic gene expression.

acetylation

histones 3

acetylation

acetylated

histone H3

interaction

SRF

myocardin-related transcriptional activator (MRTF)-A

recruitment

SRF

MRTF-A

angiogenic

angiogenesis

Neovascularization

angiogenic

angiogenic

development

vascular

repair

vascular

expression

localization

MRTF-A

inhibition

CREB-binding protein (CBP) histone acetyltransferase (HAT)

Inhibition

p38 SAPK

reduced

Inhibition

CBP HAT

reduced

Inhibition

recruitment

signaling

Actin

requires

regulation

MRTF-A

requires

regulation

CREB-binding protein histone acetyltransferase

activation

angiogenesis

induction

critical

development

critical

repair

induction

via

serum response factor

promoter

via

serum response factor

interaction

stimulated

MRTF-A(-/-) cells

induced

acetylation

mediated

CBP

acetylation

induction

induction

enhanced

induction

CBP HAT

promoter

enhanced

induction

promoter

induced

expression

activation

SRF

activation

MRTF-A

activation

CBP

regulation

respond

Smooth muscle-rich tissues

activation

mediated

RhoA

actin

mediated

RhoA

p38 stress-activated protein kinase

controls

signaling

via

associated

expression

localization

controlled

activation


Input:
Angiogenesis in platelet endothelial cell adhesion molecule-1-null mice.

Platelet endothelial cell adhesion molecule (PECAM)-1 has been previously implicated in endothelial cell migration; additionally, anti-PECAM-1 antibodies have been shown to inhibit in vivo angiogenesis. Studies were therefore performed with PECAM-1-null mice to further define the involvement of PECAM-1 in blood vessel formation. Vascularization of subcutaneous Matrigel implants as well as tumor angiogenesis were both inhibited in PECAM-1-null mice. Reciprocal bone marrow transplants that involved both wild-type and PECAM-1-deficient mice revealed that the impaired angiogenic response resulted from a loss of endothelial, but not leukocyte, PECAM-1. In vitro wound migration and single-cell motility by PECAM-1-null endothelial cells were also compromised. In addition, filopodia formation, a feature of motile cells, was inhibited in PECAM-1-null endothelial cells as well as in human endothelial cells treated with either anti-PECAM-1 antibody or PECAM-1 siRNA. Furthermore, the expression of PECAM-1 promoted filopodia formation and increased the protein expression levels of Cdc42, a Rho GTPase that is known to promote the formation of filopodia. In the developing retinal vasculature, numerous, long filamentous filopodia, emanating from endothelial cells at the tips of angiogenic sprouts, were observed in wild-type animals, but to a lesser extent in the PECAM-1-null mice. Together, these data further establish the involvement of endothelial PECAM-1 in angiogenesis and suggest that, in vivo, PECAM-1 may stimulate endothelial cell motility by promoting the formation of filopodia.

Angiogenesis

platelet endothelial cell adhesion molecule-1-null mice

angiogenesis

Vascularization

angiogenesis

tumor

angiogenic

angiogenic

sprouts

angiogenesis

formation

blood vessel

formation

filopodia

formation

filopodia

formation

filopodia

developing

retinal vasculature

formation

filopodia

platelet endothelial cell adhesion molecule-1

deficient

expression

expression

migration

endothelial cell

loss

endothelial

loss

leukocyte

motility

cell

motility

endothelial cell

inhibit

anti-PECAM-1 antibodies

angiogenesis

inhibited

Vascularization

inhibited

angiogenesis

impaired

angiogenic

loss

inhibited

formation

transplants

bone marrow

treated

endothelial cells

anti-PECAM-1 antibody

promoted

expression

formation

increased

expression

expression

promote

formation

stimulate

motility

promoting

promoting

formation

involvement

formation

involvement

angiogenesis


Input:
MEK5/ERK5 signaling modulates endothelial cell migration and focal contact turnover.

The formation of new blood vessels from pre-existing ones requires highly coordinated restructuring of endothelial cells (EC) and the surrounding extracellular matrix. Directed EC migration is a central step in this process and depends on cellular signaling cascades that initiate and control the structural rearrangements. On the basis of earlier findings that ERK5 deficiency in mouse EC results in massive defects in vessel architecture, we focused on the impact of the MEK5/ERK5 signaling pathway on EC migration. Using a retroviral gene transfer approach, we found that constitutive activation of MEK5/ERK5 signaling strongly inhibits EC migration and results in massive morphological changes. The area covered by spread EC was dramatically enlarged, accompanied by an increase in focal contacts and altered organization of actin filaments. Consequently, cells were more rigid and show reduced motility. This phenotype was most likely based on decreased focal contact turnover caused by reduced expression of p130Cas, a key player in directed cell migration. We demonstrate for the first time that ERK5 signaling not only is involved in EC survival and stress response but also controls migration and morphology of EC.

survival

EC

formation

blood vessels

deficiency

expression

migration

endothelial cell

migration

EC

migration

EC

migration

EC

spread

EC

motility

cells

migration

cell

migration

EC

defects

vessel architecture

inhibits

activation

migration

reduced

motility

reduced

expression

signaling

signaling pathway

signaling

signaling

requires

formation

restructuring

results

deficiency

defects

activation

signaling

modulates

signaling

migration

impact

signaling pathway

migration

involved

signaling

survival

controls

signaling

migration

controls

signaling

EC

restructuring

endothelial cells

restructuring

extracellular matrix

organization

filaments


Input:
EPOX inhibits angiogenesis by degradation of Mcl-1 through ERK inactivation.

PURPOSE: Antiangiogenic therapy is considered as an effective strategy for controlling the growth and metastasis of tumors. Among a myriad of biological activities described for xanthone derivatives, the anticancer activity is quite remarkable, but the molecular mechanism is not clearly resolved. In the present study, we investigated the antiangiogenic mechanism of 3,6-di(2,3-epoxypropoxy)xanthone (EPOX), a novel Mcl-1 targeting drug. EXPERIMENTAL DESIGN: To evaluate the antiangiogenic activity of EPOX, we did cell viability, cell cycle, tube formation assay in vitro, and Matrigel plug assay in vivo. To evaluate the effect of EPOX on the endothelial signaling pathway, we did immunoblotting, immunoprecipitation, and immunofluorescence analysis. Intracellular glutathione levels were determined with the use of monochlorobimane, a glutathione-specific probe. RESULTS: EPOX induced endothelial cell apoptosis in association with proteasome-dependent Mcl-1 degradation. Down-regulation of Mcl-1 resulted in an increase in Mcl-1-free Bim, activation of Bax, and then signaling of mitochondria-mediated apoptosis. Additionally, glutathione depletion and extracellular signal-regulated kinase (ERK) inactivation was observed in EPOX-treated cells. Glutathione supplementation reversed the inhibitory effects of EPOX on ERK, which increases the phosphorylation of Mcl-1 at T(163.) Overexpression of mitogen-activated protein/ERK kinase (MEK) partially reversed the effect of EPOX on Mcl-1 dephosphorylation, ubiquitination, and degradation, further implicating ERK in the regulation of Mcl-1 stability. CONCLUSIONS: This study provides evidence that EPOX induces glutathione depletion, ERK inactivation, and Mcl-1 degradation on endothelial cells, which leads to inhibition of angiogenesis. Our results suggest that EPOX is a novel antiangiogenic agent, making it a promising lead compound for further development in the treatment of angiogenesis-related pathologies.

angiogenesis

angiogenic

angiogenic

angiogenic

angiogenesis

angiogenic

angiogenesis

degradation

degradation

degradation

degradation

apoptosis

endothelial cell

dephosphorylation

formation

tube

Overexpression

mitogen-activated protein/ERK kinase

growth

tumors

metastasis

tumors

inhibits

EPOX

angiogenesis

inactivation

ERK

therapy

angiogenic

activity

xanthone derivatives

cancer

mechanism

angiogenic

activity

angiogenic

EPOX

Down-regulation

depletion

glutathione

inactivation

extracellular signal-regulated kinase

reversed

supplementation

inhibitory effects

inhibitory effects

EPOX

ERK

reversed

Overexpression

effect

depletion

glutathione

inactivation

ERK

inhibition

angiogenesis

endothelial signaling pathway

phosphorylation

treated

EPOX

cells

supplementation

Glutathione

through

degradation

inactivation

induced

EPOX

apoptosis

dependent

degradation

resulted

Down-regulation

increase

resulted

Down-regulation

activation

increase

Bim

activation

Bax

increases

ERK

phosphorylation

induces

EPOX

depletion

induces

EPOX

inactivation

induces

EPOX

degradation

leads

induces

inhibition

controlling

therapy

growth

controlling

therapy

metastasis

targeting

effect

EPOX

endothelial signaling pathway

association

induced

degradation

free

Bim

effect

EPOX

dephosphorylation

effect

EPOX

ubiquitination

effect

EPOX

degradation

regulation

ERK

ubiquitination


Input:
Targeting angiogenesis: progress with anti-VEGF treatment with large molecules.

Angiogenesis--one of the hallmarks of cancer--has emerged as a valid therapeutic target in oncology. The VEGF system represents a key mediator of tumor-initiated angiogenesis and the first target of antiangiogenesis agents introduced in clinical practice. Although anti-VEGF therapies have clearly demonstrated antitumor efficacy in various malignancies, especially when combined with conventional cytotoxic chemotherapy, their mechanism of action is not fully understood. This Review will discuss the rationale for using antiangiogenic compounds and will focus on large molecules, such as antibodies, that target the VEGF system. Clinical data on bevacizumab is discussed in detail. Predictive markers for anti-VEGF agents have not yet been identified and questions regarding the usefulness of bevacizumab in the adjuvant setting as well as its continued use beyond progression remain unanswered, in spite of negative data on bevacizumab in treating patients with adjuvant colon cancer. Nonetheless, anti-VEGF therapy has enhanced the arsenal of anticancer therapies and has provided new insights into the biology of malignancy.

angiogenesis

Angiogenesis

angiogenesis

angiogenesis

angiogenic

efficacy

therapies

tumor

system

VEGF

system

VEGF

therapies

anti-VEGF

treating

bevacizumab

patients

initiated

tumor

angiogenesis


Input:
[Ambiguity role of neutrophils in oncogenesis]

The review is focused on the participation of polymorphonuclear granulocytes (neutrophils) in development and spreading of a tumor. We consider both the well known functions of neutrophils (degranulation, production of reactive oxygen species (ROS)) and the recently shown one (presentation of an antigene). The special attention is focused on the ambiguity of the neutrophil role in oncogenesis. The dominant view is that neutrophils display exclusively antitumor properties. The update information testifies about protumoral activity of neutrophils: they migrate to a tumor and promote angiogenesis and metastasis at late stages of the tumor. It is interesting that certain components of neutrophil cytotoxic arsenal (ROS, cytokines, specific enzymes) participate both in antitumoral defenses of an organism and protumoral activity.

angiogenesis

tumor

development

tumor

spreading

tumor

migrate

neutrophils

tumor

metastasis

tumor

properties

neutrophils

tumor

promote

neutrophils

angiogenesis

promote

neutrophils

metastasis

production

reactive oxygen species


Input:
Endothelial cell migration and vascular endothelial growth factor expression are the result of loss of breast tissue polarity.

Recruiting a new blood supply is a rate-limiting step in tumor progression. In a three-dimensional model of breast carcinogenesis, disorganized, proliferative transformed breast epithelial cells express significantly higher expression of angiogenic genes compared with their polarized, growth-arrested nonmalignant counterparts. Elevated vascular endothelial growth factor (VEGF) secretion by malignant cells enhanced recruitment of endothelial cells (EC) in heterotypic cocultures. Significantly, phenotypic reversion of malignant cells via reexpression of HoxD10, which is lost in malignant progression, significantly attenuated VEGF expression in a hypoxia-inducible factor 1alpha-independent fashion and reduced EC migration. This was due primarily to restoring polarity: forced proliferation of polarized, nonmalignant cells did not induce VEGF expression and EC recruitment, whereas disrupting the architecture of growth-arrested, reverted cells did. These data show that disrupting cytostructure activates the angiogenic switch even in the absence of proliferation and/or hypoxia and restoring organization of malignant clusters reduces VEGF expression and EC activation to levels found in quiescent nonmalignant epithelium. These data confirm the importance of tissue architecture and polarity in malignant progression.

angiogenic

proliferation

nonmalignant cells

progression

tumor

transformed

breast epithelial cells

phenotypic reversion

malignant cells

progression

malignant

expression

vascular endothelial growth factor

reexpression

expression

VEGF

expression

VEGF

expression

VEGF

migration

Endothelial cell

Recruiting

blood

secretion

vascular endothelial growth factor

malignant cells

recruitment

endothelial cells

migration

EC

recruitment

EC

attenuated

phenotypic reversion

expression

reduced

phenotypic reversion

migration

disrupting

cytostructure

reduces

expression

reduces

activation

Elevated

secretion

enhanced

Elevated

recruitment

forced

proliferation

induce

proliferation

expression

induce

proliferation

recruitment

activates

disrupting

angiogenic

activation

EC

independent

attenuated

hypoxia-inducible factor 1alpha


Input:
Metronomic 5-fluorouracil, oxaliplatin and irinotecan in colorectal cancer.

Metronomic chemotherapy (the frequent, long term, low dose administration of chemotherapeutic drugs) is a promising therapy because it enhances the anti-endothelial activity of conventional chemotherapeutics, but with lower or no toxic effects compared to maximum tolerated dose administration. The aims of the present study were to compare, in vitro and in vivo, the antiangiogenic and antitumor activities of metronomic irinotecan (CPT-11), oxaliplatin (L-OHP) and 5-fluorouracil (5-FU) in colorectal cancer and to investigate the metronomic combination of these drugs. In vitro cell proliferation, combination studies and vascular endothelial growth factor (VEGF) secretion analyses were performed on endothelial (HMVEC-d) and colorectal cancer (HT-29) cells exposed for 144 h to metronomic concentrations of SN-38, the active metabolite of CPT-11, L-OHP and 5-FU. HT-29 human colorectal cancer xenograft model was used and tumour growth, microvessel density and VEGF quantification were performed in tumours after the administration of metronomic CPT-11, L-OHP, 5-FU and their simultaneous combination. Low concentrations of SN-38, but not 5-FU and L-OHP, preferentially inhibited endothelial cell proliferation. Simultaneous and continuous exposure of HT-29 and HMVEC-d cells to low concentrations SN-38+L-OHP+5-FU for 144 h showed a strong antagonism and an unfavorable dose-reduction index. Moreover, the ternary combination resulted in a significant increase of VEGF secretion in HT-29 cancer cells. In a xenograft model metronomic CPT-11, but not 5-FU and L-OHP, significantly inhibits HT-29 tumor growth and microvessel density in the absence of toxicity. On the contrary, metronomic 5-FU+L-OHP+CPT-11 therapy did not affect the microvascular count. The metronomic concept might not universally apply to every cytotoxic drug in colorectal cancer and metronomic combination regimens should be used with caution.

angiogenic

proliferation

cell

proliferation

endothelial cell

growth

tumour

tumor growth

HT-29 tumor

secretion

vascular endothelial growth factor

secretion

VEGF

activity

endothelial

activities

angiogenic

activities

tumor

inhibited

proliferation

inhibited

proliferation

inhibited

L-OHP

proliferation

inhibits

tumor growth

inhibits

microvessel

inhibits

tumor growth

inhibits

microvessel

inhibits

L-OHP

tumor growth

inhibits

L-OHP

microvessel

exposed

endothelial (HMVEC-d)

exposed

colorectal cancer (HT-29) cells

exposed

endothelial (HMVEC-d)

exposed

colorectal cancer (HT-29) cells

exposed

endothelial (HMVEC-d)

L-OHP

exposed

colorectal cancer (HT-29) cells

L-OHP

exposed

endothelial (HMVEC-d)

exposed

colorectal cancer (HT-29) cells

exposure

L-OHP

exposure

HMVEC-d cells

L-OHP

therapy

L-OHP

enhances

activity

increase

secretion

affect

therapy

microvascular


Input:
A key role for the integrin alpha2beta1 in experimental and developmental angiogenesis.

The alpha2beta1 integrin receptor plays a key role in angiogenesis. Here we investigated the effects of small molecule inhibitors (SMIs) designed to disrupt integrin alpha2 I or beta1 I-like domain function on angiogenesis. In unchallenged endothelial cells, fibrillar collagen induced robust capillary morphogenesis. In contrast, tube formation was significantly reduced by SMI496, a beta1 I-like domain inhibitor and by function-blocking anti-alpha2beta1 but not -alpha1beta1 antibodies. Endothelial cells bound fluorescein-labeled collagen I fibrils, an interaction specifically inhibited by SMI496. Moreover, SMI496 caused cell retraction and cytoskeletal collapse of endothelial cells as well as delayed endothelial cell wound healing. SMI activities were examined in vivo by supplementing the growth medium of zebrafish embryos expressing green fluorescent protein under the control of the vascular endothelial growth factor receptor-2 promoter. SMI496, but not a control compound, interfered with angiogenesis in vivo by reversibly inhibiting sprouting from the axial vessels. We further characterized zebrafish alpha2 integrin and discovered that this integrin is highly conserved, especially the I domain. Notably, a similar vascular phenotype was induced by morpholino-mediated knockdown of the integrin alpha2 subunit. By live videomicroscopy, we confirmed that the vessels were largely nonfunctional in the absence of alpha2beta1 integrin. Collectively, our results provide strong biochemical and genetic evidence of a central role for alpha2beta1 integrin in experimental and developmental angiogenesis.

bound

Endothelial cells

collagen I fibrils

angiogenesis

angiogenesis

angiogenesis

angiogenesis

angiogenesis

cytoskeletal collapse

endothelial cells

morphogenesis

capillary

formation

tube

sprouting

axial vessels

expressing

green fluorescent protein

reduced

formation

reduced

formation

reduced

formation

-alpha1beta1 antibodies

inhibited

bound

delayed

endothelial cell

interfered

angiogenesis

inhibiting

inhibiting

sprouting

unchallenged

endothelial cells

labeled

collagen I fibrils

knockdown

integrin alpha2

induced

fibrillar collagen

morphogenesis

caused

cytoskeletal collapse

plays a key role

alpha2beta1 integrin receptor

angiogenesis

control

expressing

vascular endothelial growth factor receptor-2

promoter

central role

alpha2beta1 integrin

angiogenesis


Input:
Islet endothelial activation and oxidative stress gene expression is reduced by IL-1Ra treatment in the type 2 diabetic GK rat.

BACKGROUND: Inflammation followed by fibrosis is a component of islet dysfunction in both rodent and human type 2 diabetes. Because islet inflammation may originate from endothelial cells, we assessed the expression of selected genes involved in endothelial cell activation in islets from a spontaneous model of type 2 diabetes, the Goto-Kakizaki (GK) rat. We also examined islet endotheliuml/oxidative stress (OS)/inflammation-related gene expression, islet vascularization and fibrosis after treatment with the interleukin-1 (IL-1) receptor antagonist (IL-1Ra). METHODOLOGY/PRINCIPAL FINDINGS: Gene expression was analyzed by quantitative RT-PCR on islets isolated from 10-week-old diabetic GK and control Wistar rats. Furthermore, GK rats were treated s.c twice daily with IL-1Ra (Kineret, Amgen, 100 mg/kg/day) or saline, from 4 weeks of age onwards (onset of diabetes). Four weeks later, islet gene analysis and pancreas immunochemistry were performed. Thirty-two genes were selected encoding molecules involved in endothelial cell activation, particularly fibrinolysis, vascular tone, OS, angiogenesis and also inflammation. All genes except those encoding angiotensinogen and epoxide hydrolase (that were decreased), and 12-lipoxygenase and vascular endothelial growth factor (that showed no change), were significantly up-regulated in GK islets. After IL-1Ra treatment of GK rats in vivo, most selected genes implied in endothelium/OS/immune cells/fibrosis were significantly down-regulated. IL-1Ra also improved islet vascularization, reduced fibrosis and ameliorated glycemia. CONCLUSIONS/SIGNIFICANCE: GK rat islets have increased mRNA expression of markers of early islet endothelial cell activation, possibly triggered by several metabolic factors, and also some defense mechanisms. The beneficial effect of IL-1Ra on most islet endothelial/OS/immune cells/fibrosis parameters analyzed highlights a major endothelial-related role for IL-1 in GK islet alterations. Thus, metabolically-altered islet endothelium might affect the beta-cell microenvironment and contribute to progressive type 2 diabetic beta-cell dysfunction in GK rats. Counteracting islet endothelial cell inflammation might be one way to ameliorate/prevent beta-cell dysfunction in type 2 diabetes.

vascularization

islet

angiogenesis

vascularization

islet

dysfunction

islet

alterations

islet

reduced

activation

treatment

decreased

angiotensinogen

decreased

epoxide hydrolase

treatment

GK rat

treatment

interleukin-1 (IL-1) receptor antagonist

isolated

islets

treated

GK rats

treated

GK rats

saline

treatment

GK rats

activation

Islet endothelial

activation

endothelial cell

activation

endothelial cell

up-regulated

angiotensinogen

up-regulated

epoxide hydrolase

up-regulated

up-regulated

vascular endothelial growth factor

improved

vascularization

activation

islet endothelial cell

beneficial effect

islet endothelial

beneficial effect

immune cells

showed no change

showed no change

vascular endothelial growth factor

role

alterations

altered

islet endothelium

affect

altered

beta-cell


Input:
AVE8062: a new combretastatin derivative vascular disrupting agent.

Angiogenesis has an essential role in promoting and supporting tumor growth and it is an important therapeutic target. The tumor vascular network is the result of pro-angiogenic and inhibitory factors as well as of the interaction between endothelial cells and extracellular matrix. Different antiangiogenic therapeutics have been developed to improve tumor control through vascular-targeting agents (VTA). VTAs can be divided into two groups: antiangiogenic agents and vascular-disrupting agents (VDAs). VTAs inhibit specific factors required to induce and direct the angiogenic process, with major activity against small tumor masses and at the tumor periphery, encompassing monoclonal antibodies and small molecules inhibitors of the tyrosine kinase domain of the VEGF receptor. VDAs specifically target and destroy well-established tumor vessels with ischemia and destruction of large masses with central hemorrhagic necrosis and survival of a thin peripheral tumor layer. VDAs can be divided into biologics, such as ligand-based, and small-molecule agents; this second group includes small-molecule VDAs like flavonoids, such as 5,6-dimethylxanthenone-4-acetic acid (DMXAA), and microtubule-destabilizing agents. In this review we will discuss the mechanism of action, as well as the preclinical and clinical results, of one of the most promising antitubulin agents: the combretastatin A4-phosphate derivative, AVE8062A.

interaction

endothelial cells

extracellular matrix

Angiogenesis

angiogenic

angiogenic

angiogenic

angiogenic

disrupting

vascular

destruction

masses

necrosis

masses

survival

peripheral tumor layer

established

tumor vessels

growth

tumor

destroy

VDAs

tumor vessels

promoting

growth

supporting

growth

improve

control

vascular-targeting agents

has an essential role

Angiogenesis

promoting

has an essential role

Angiogenesis

supporting

control

tumor

target

VDAs

tumor vessels


Input:
Targeting TNF for Treatment of Cancer and Autoimmunity.

Tumor necrosis factor-alpha (TNF-alpha) was first isolated two decades ago as a macrophageproduced protein that can effectively kill tumor cells. TNF-alpha is also an essential component of the immune system and is required for hematopoiesis, for protection from bacterial infection and for immune cell-mediated cytotoxicity. Extensive research, however, has revealed that TNF-alpha is one of the major players in tumor initiation, proliferation, invasion, angiogenesis and metastasis. The proinflammatory activities link TNF-alpha with a wide variety of autoimmune diseases, including psoriasis, inflammatory bowel disease, rheumatoid arthritis, systemic sclerosis, systemic lupus erythematosus, multiple sclerosis, diabetes and ankylosing spondylitis. Systemic inhibitors of TNF such as etanercept (Enbrel) (a soluble TNF receptor) and infliximab (Remicade) and adalimumab (Humira) (anti-TNF antibodies) have been approved for the treatment inflammatory bowel disease, psoriasis and rheumatoid arthritis. These drugs, however, exhibit severe side effects and are expensive. Hence orally active blockers of TNF-alpha that are safe, efficacious and inexpensive are urgently needed. Numerous products from fruits, vegetable and traditional medicinal plants have been described which can suppress TNF expression and TNF signaling but their clinical potential is yet uncertain.

angiogenesis

tumor

kill

tumor cells

expression

TNF

proliferation

tumor

invasion

tumor

metastasis

tumor

suppress

expression

suppress

signaling

signaling

TNF

Treatment

TNF

Cancer

treatment

etanercept

treatment

infliximab

treatment

adalimumab

kill

Tumor necrosis factor-alpha

kill

initiation

tumor

major players

TNF-alpha

initiation

major players

TNF-alpha

proliferation

major players

TNF-alpha

invasion

major players

TNF-alpha

angiogenesis

major players

TNF-alpha

metastasis


Input:
Expression of basic fibroblast growth factor in normal human tissues.

The distribution of basic fibroblast growth factor (bFGF) was studied immunohistochemically in fresh frozen sections of normal human tissues. Immunodetection was performed with a specific anti-bFGF mouse monoclonal antibody that was found to react with recombinant human bFGF in Western blot analysis, and to specifically neutralize the mitogenic activity of bFGF on bovine vascular endothelial cells. Expression of bFGF on normal human tissues was ubiquitously detected in the basement membranes of all size blood vessels, but was not found in epidermal or epithelial basement membranes of a variety of tissues tested. Intensity and patterns of localization in blood vessels was consistent in various tissues, but varied among different regions of the vascular bed. Whereas homogeneous and intense immunoreactivity were observed in large and intermediate size blood vessels, heterogeneity of expression was found in capillaries. The most intense immunoreactivity was observed in branching capillaries. Endothelial cell staining was heterogeneous and varied in different regions. Strong staining for bFGF was also found in cardiac muscle fibers, smooth muscle cells of mid-size blood vessels, the gut and the myometrium, in central nervous system neurons and cerebellar Purkinje cells, and on epithelial cells of the bronchi, colon, endometrium, and sweat gland ducts of the skin. The presence of bFGF in the extracellular compartment of a diverse variety of organs may play a role in angiogenesis. However, the function of bFGF in parenchymal cells remains to be determined.

angiogenesis

branching

capillaries

Expression

basic fibroblast growth factor

Expression

bFGF

expression

bFGF

distribution

basic fibroblast growth factor

ubiquitously detected

bFGF

basement membranes

ubiquitously detected

bFGF

blood vessels

localization

bFGF

blood vessels

localization

bFGF

vascular bed

found

bFGF

cardiac muscle fibers

found

bFGF

smooth muscle cells

found

bFGF

gut

found

bFGF

myometrium

found

bFGF

central nervous system neurons

found

bFGF

cerebellar Purkinje cells

found

bFGF

bronchi

found

bFGF

colon

found

bFGF

endometrium

found

bFGF

sweat gland ducts

found

bFGF

skin

presence

bFGF

extracellular compartment

neutralize

anti-bFGF mouse monoclonal antibody

bFGF

Immunodetection

anti-bFGF mouse monoclonal antibody

bFGF

play a role

bFGF

angiogenesis


Input:
Retinal revascularisation in diabetic retinopathy.

The case history of a 33-year-old diabetic patient who has had diabetes for 24 years is presented. When first seen in 1975 he had bilateral proliferative retinopathy with new vessels in the retinal periphery. He had large areas of capillary non-perfusion lateral to the macula in the right eye associated with the new vessels. Nine years later, after extensive repeated photocoagulation, revascularisation of large areas previously not perfused were seen. The vessels are in the plane of the retina and do not have the appearance of new vessels.

revascularisation

Retinal

revascularisation

appearance

vessels


Input:
Mast cells and tumors. The specific enhancement of tumor proliferation in vitro.

Mast cells were found to be unique among the peritoneal leukocytes by virtue of their capacity to enhance profoundly the proliferation of a variety of tumors in vitro. This phenomenon occurs at mast cell/tumor ratios which reflect the stoichiometry of host cell/tumor relationships in vivo. The growth factor was found to reside in mast cell granules and was identified as heparin by sequential purification and enzymatic degradation. This cellular interaction was tumor-specific, although isolated granules could enhance fibroblast proliferation. The findings are discussed in relation to previous morphologic studies, reports of in vitro mast-cell-mediated tumor cytotoxicity, and the role of mast cells in angiogenesis and connective tissue proliferation.

angiogenesis

proliferation

fibroblast

proliferation

tumor

proliferation

tumors

proliferation

connective tissue

enhancement

proliferation

enhance

Mast cells

proliferation

enhance

proliferation

mediated

mast-cell

tumor

role

mast cells

angiogenesis

role

mast cells

proliferation


Input:
Subretinal neovascularization following rubella retinopathy.

A 17-year-old girl and an 11-year-old girl with rubella retinopathy had decreased vision in one eye secondary to subretinal neovascularization and hemorrhage. In both cases a disciform scar with permanent decrease in central vision resulted.

neovascularization

neovascularization


Input:
VEGF165 expressed by a replication-deficient recombinant adenovirus vector induces angiogenesis in vivo.

To evaluate the concept that localized delivery of angiogenic factors via virus-mediated gene transfer may be useful in the treatment of ischemic disorders, the replication-deficient adenovirus (Ad) vector AdCMV.VEGF165 (where CMV is cytomegalovirus and VEGF is vascular endothelial growth factor) containing the cDNA for human VEGF165, a secreted endothelial cell-specific angiogenic growth factor, was constructed. Human umbilical vein endothelial cells (HUVECs) and rat aorta smooth muscle cells (RASMCs) infected with AdCMV.VEGF165 (5 and 20 plaque-forming units [pfu] per cell) demonstrated VEGF mRNA expression and protein secretion into the supernatant. Furthermore, the conditioned medium from these cells enhanced vascular permeability in vivo. In contrast, neither VEGF mRNA nor secreted protein was found in uninfected HUVECs or RASMCs or in cells infected with the control vector AdCMV.beta gal (where beta gal is beta-galactosidase). Assessment of starved HUVECs at 14 days demonstrated sixfold more cells for AdCMV.VEGF165-infected HUVECs (20 pfu per cell) than for either infected or uninfected control cells. RASMC proliferation was unaffected by infection with AdCMV.VEGF165. When plated in 2% serum on dishes precoated with reconstituted basement membrane (Matrigel), HUVECs infected with AdCMV.VEGF165 (20 pfu per cell) differentiated into capillary-like structures. Under similar conditions, both uninfected HUVECs and HUVECs infected with AdCMV.beta gal did not differentiate. To evaluate the ability of AdCMV.VEGF165 to function in vivo, either AdCMV. VEGF165 or AdCMV.beta gal (2 x 10(10) pfu) was resuspended in 0.5 mL Matrigel and injected subcutaneously into mice. Immunohistochemical staining demonstrated VEGF in the tissues surrounding the Matrigel plugs containing AdCMV.VEGF165 up to 3 weeks after injection, whereas no VEGF was found in the control plugs with AdCMV.beta gal. Two weeks after injection, there was histological evidence of neovascularization in the tissues surrounding the Matrigel containing AdCMV.VEGF165, whereas no significant angiogenesis was observed in response to AdCMV.beta gal. Furthermore, the Matrigel plugs with AdCMV.VEGF165 demonstrated hemoglobin content fourfold higher than the plugs with AdCMV.beta gal. Together, these in vitro and in vivo studies are consistent with the concept that Ad vectors may provide a useful strategy for efficient local delivery of VEGF165 in the treatment of ischemic diseases.

angiogenesis

angiogenic

angiogenic

neovascularization

tissues

angiogenesis

tissues

proliferation

RASMC

differentiated

HUVECs

differentiate

HUVECs

expressed

secretion

VEGF

secreted

VEGF

infected

Human umbilical vein endothelial cells

infected

rat aorta smooth muscle cells

uninfected

HUVECs

uninfected

RASMCs

infected

cells

AdCMV.beta gal

infected

HUVECs

infected

cells

uninfected

cells

infection

plated

basement membrane

HUVECs

infected

HUVECs

uninfected

HUVECs

infected

HUVECs

AdCMV.beta gal

resuspended

AdCMV.beta gal

injected

AdCMV.beta gal

mice

Immunohistochemical staining

VEGF

injection

induces

expressed

angiogenesis

enhanced

vascular

unaffected

proliferation

infection

response

angiogenesis

AdCMV.beta gal

expression

VEGF


Input:
Effects of doxycycline on in vitro growth, migration, and gelatinase activity of breast carcinoma cells.

Metastatic disease is one of the major causes of death from cancer in human beings. Several enzyme systems have been implicated in the metastatic process, but the metalloproteinases (MPs) appear to be the major group involved in most instances of neoplastic invasion. Increased MP activity has been correlated with the metastatic potential of many cancers, including breast cancer. MPs also play a role in tumor angiogenesis. Tetracyclines are antimicrobial agents that can suppress MP activity in a variety of tissues, including gingiva, bone, and cartilage. Several reports have indicated that tetracyclines can suppress tumor MPs as well. A synthetic tetracycline, doxycycline, inhibits migration of human MDA-MB-435 breast adenocarcinoma cells through a reconstituted basement membrane (Matrigel), an assay used as an in vitro surrogate for the in vivo process of tumor invasion through basement membranes. Additionally, doxycycline diminishes the proliferation of this breast cancer cell line and also decreases its gelatinolytic activity, as determined by gel zymography.

angiogenesis

tumor

growth

breast carcinoma cells

proliferation

breast cancer cell line

death

human

migration

breast carcinoma cells

migration

MDA-MB-435 breast adenocarcinoma cells

invasion

tumor

suppress

Tetracyclines

MP

suppress

tetracyclines

MPs

inhibits

tetracycline

migration

inhibits

doxycycline

migration

diminishes

doxycycline

proliferation

Increased

MP

Effects

doxycycline

growth

Effects

doxycycline

migration

play a role

MPs

angiogenesis

reconstituted

basement membrane


Input:
Stimulation of endothelial cell migration by vascular permeability factor/vascular endothelial growth factor through cooperative mechanisms involving the alphavbeta3 integrin, osteopontin, and thrombin.

We have identified several mechanisms by which the angiogenic cytokine vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) likely regulates endothelial cells (EC) migration. VPF/VEGF induced dermal microvascular EC expression of mRNAs encoding the alphav and beta3 integrin subunits resulting in increased levels of the alphavbeta3 heterodimer at the cell surface, and VPF/VEGF also induced mRNA encoding osteopontin (OPN), an alphavbeta3 ligand. OPN promoted EC migration in vitro; and VPF/VEGF induction of alphavbeta3 was accompanied by increased EC migration toward OPN. Because thrombin cleavage of OPN results in substantial enhancement of OPN's adhesive properties, and because VPF/VEGF promotes increased microvascular permeability leading to activation of the extrinsic coagulation pathway, we also investigated whether VPF/VEGF facilitates thrombin cleavage of OPN in vivo. Consistent with this hypothesis, co-injection of VPF/VEGF together with OPN resulted in rapid cleavage of OPN by endogenous thrombin. Furthermore, in comparison with native OPN, thrombin-cleaved OPN stimulated a greater rate of EC migration in vitro, which was additive to the increased migration associated with induction of alpha v beta 3. Thus, these data demonstrate cooperative mechanisms for VPF/VEGF regulation of EC migration involving the alphavbeta3 integrin, the alphavbeta3 ligand OPN, and thrombin cleavage of OPN. These findings also illustrate an operational link between VPF/VEGF induction of EC gene expression and VPF/VEGF enhancement of microvascular permeability, suggesting that these distinct biological activities may act accordingly to stimulate EC migration during angiogenesis.

angiogenic

angiogenesis

cleavage

OPN

cleavage

OPN

cleavage

OPN

cleaved

OPN

cleavage

OPN

migration

endothelial cell

migration

endothelial cells

migration

EC

migration

EC

migration

EC

migration

EC

migration

EC

extrinsic coagulation pathway

co-injection

VPF

OPN

Stimulation

migration

vascular permeability factor

induced

VPF

expression

increased

alphav

increased

beta3 integrin subunits

induced

VPF

osteopontin

promoted

OPN

migration

induction

VPF

increased

migration

cleavage

thrombin

cleavage

enhancement

OPN

promotes

VPF

increased

increased

microvascular

activation

extrinsic coagulation pathway

cleavage

thrombin

cleavage

cleavage

cleavage

thrombin

cleaved

thrombin

cleaved

stimulated

OPN

migration

stimulated

OPN

migration

induction

alpha v beta 3

cleavage

thrombin

cleavage

enhancement

VPF

microvascular

stimulate

migration

regulates

vascular permeability factor

migration

facilitates

VPF

cleavage

regulation

VPF

migration

expression

alphav


Input:
Gene transfer of naked DNA encoding for three isoforms of vascular endothelial growth factor stimulates collateral development in vivo.

Vascular endothelial growth factor (VEGF) is a naturally secreted endothelial cell-specific mitogen. We investigated the hypothesis that naked DNA encoding for VEGF could be used in a strategy of arterial gene therapy to stimulate collateral artery development. Plasmid DNA encoding each of the three principal human VEGF isoforms (phVEGF121, phVEGF165, or phVEGF189) was applied to the hydrogel polymer coating of an angioplasty balloon and delivered percutaneously to one iliac artery of rabbits with operatively induced hindlimb ischemia. Compared with control animals transfected with LacZ, site-specific transfection of phVEGF resulted in augmented collateral vessel development documented by serial angiography, and improvement in calf blood pressure ratio (ischemic to normal limb), resting and maximum blood flow, and capillary to myocyte ratio. Similar results were obtained with phVEGF121, phVEGF165, and phVEGF189, which suggests that these isoforms are biologically equivalent with respect to in vivo angiogenesis. The fact that viral or other adjunctive vectors were not required further suggests that secreted gene products may have potential therapeutic utility even when the number of successfully transfected cells remains low. Arterial gene transfer of naked DNA encoding for a secreted angiogenic cytokine, thus, represents a potential alternative to recombinant protein administration for stimulating collateral vessel development.

angiogenesis

angiogenic

development

collateral

development

collateral artery

development

collateral vessel

development

collateral vessel

secreted

Vascular endothelial growth factor

endothelial cell

arterial gene therapy

VEGF

transfected

LacZ

site-specific transfection

phVEGF

stimulates

vascular endothelial growth factor

development

stimulate

VEGF

development

resulted

site-specific transfection

development

stimulating

development


Input:
Induction of Ets-1 in endothelial cells during reendothelialization after denuding injury.

Ets-1, a transcription factor, is induced in endothelial cells (ECs) during angiogenesis. Here, we investigated the expression of Ets-1 during reendothelialization. When a confluent monolayer of human umbilical vein endothelial cell line, ECV304, was denuded, ECV304 at the wound edge expressed Ets-1. An immunohistochemical analysis revealed that Ets-1 accumulated in migrating cells at the wound edge and returned to basal level when reendothelialization was accomplished. This induction of Ets-1 could be reproduced in in vivo denudation of rat aortic endothelium by a balloon catheter. The induction of Ets-1 in ECs after denudation was regulated transcriptionally, and humeral factors released from injured ECs might not be responsible. Mitogen-activated protein kinase (MAPK) activities were investigated to explore the mechanism of this induction. Although extracellular signal-regulated protein kinase 1/2 (ERK1/2), c-Jun N-terminal kinase 1 (JNK1), and p38 were activated after denudation, the activation of ERK1 and p38 was more rapid and prominent. PD98059, a specific MAPK/ERK kinase (MEK) 1 inhibitor, did not affect the induction of ets-1 mRNA, whereas SB203580, a specific p38 inhibitor, almost completely abrogated its induction. These results indicate that Ets-1 is induced in ECs after denudation through activation of p38. This induction of Ets-1 may be relevant for reendothelialization by regulating the expression of certain genes.

reendothelialization

angiogenesis

reendothelialization

reendothelialization

reendothelialization

expression

expressed

released

humeral factors

ECs

abrogated

induction

denuded

human umbilical vein endothelial cell line

denuded

denudation

aortic endothelium

denudation

ECs

denudation

ECs

Induction

induced

accumulated

induction

reproduced

induction

denudation

induction

induction

activated

activated

c-Jun N-terminal kinase 1

activated

activated

extracellular signal-regulated protein kinase 1

activation

activation

induction

induction

induced

activation

activation

induction

regulated

induction

affect

induction


Input:
Store-operated calcium entry promotes shape change in pulmonary endothelial cells expressing Trp1.

Activation of Ca2+ entry is known to produce endothelial cell shape change, leading to increased permeability, leukocyte migration, and initiation of angiogenesis in conduit-vessel endothelial cells. The mode of Ca2+ entry regulating cell shape is unknown. We hypothesized that activation of store-operated Ca2+ channels (SOCs) is sufficient to promote cell shape change necessary for these processes. SOC activation in rat pulmonary arterial endothelial cells increased free cytosolic Ca2+ that was dependent on a membrane current having a net inward component of 5.45 +/- 0.90 pA/pF at -80 mV. Changes in endothelial cell shape accompanied SOC activation and were dependent on Ca2+ entry-induced reconfiguration of peripheral (cortical) filamentous actin (F-actin). Because the identity of pulmonary endothelial SOCs is unknown, but mammalian homologues of the Drosophila melanogaster transient receptor potential (trp) gene have been proposed to form Ca2+ entry channels in nonexcitable cells, we performed RT-PCR using Trp oligonucleotide primers in both rat and human pulmonary arterial endothelial cells. Both cell types were found to express Trp1, but neither expressed Trp3 nor Trp6. Our study indicates that 1) Ca2+ entry in pulmonary endothelial cells through SOCs produces cell shape change that is dependent on site-specific rearrangement of the microfilamentous cytoskeleton and 2) Trp1 may be a component of pulmonary endothelial SOCs.

angiogenesis

shape change

pulmonary endothelial cells

shape change

endothelial cell

shape change

cell

Changes

endothelial cell

shape change

cell

expressing

express

expressed

expressed

entry

calcium

entry

migration

leukocyte

entry

entry

entry

entry

promotes

entry

shape change

Activation

entry

initiation

angiogenesis

increased

dependent

increased

membrane

dependent

Changes

reconfiguration

induced

entry

reconfiguration

produce

Activation

shape change

produce

Activation

migration

produce

Activation

initiation

reconfiguration

peripheral (cortical) filamentous actin

produces

entry

shape change


Input:
Effect of U-995, a potent shark cartilage-derived angiogenesis inhibitor, on anti-angiogenesis and anti-tumor activities.

BACKGROUND: A potent angiogenesis inhibitor, U-995, has been purified from the cartilage of the blue shark (Prionace glauca). U-995 is composed of two single peptides with molecular mass of 10 and 14 kDa, respectively. MATERIALS AND METHODS: U-995 was designed to study human umbilical vein endothelial cell (HUVEC) migration and proliferation in vitro and angiogenesis induced by TNF alpha in chicken chorioallantoic membrane (CAM). Furthermore, we determined the ability of U-995 to inhibiting tumor cell growth and metastasis. RESULTS: U-995 (15 and 30 micrograms/ml) markedly inhibited HUVEC migration and, at 15-50 micrograms/ml produced a dose-dependent decline in [3H]-thymidine incorporation. 30 and 50 micrograms/ml of U-995, when added to TNF alpha-induced angiogenesis caused discontinuous and disrupted blood vessels. Moreover, U-995 (30 micrograms/ml) markedly prevented collagenase-induced collagenolysis. In addition, when 200 micrograms U-995 was injected i.p. into mice it suppressed sarcoma-180 cell growth and B16-F10 mouse melanoma cell metastasis in vivo. CONCLUSIONS: These results suggest that the anti-angiogenic effects of U-995 may be be due to interference with the proliferation and migration of HUVECs as well as inhibition of collagenolysis, thereby leading to inhibition of both angiogenesis and tumor cell growth.

angiogenesis

angiogenesis

angiogenesis

angiogenesis

angiogenesis

collagenolysis

angiogenic

angiogenesis

disrupted

blood vessels

proliferation

human umbilical vein endothelial cell

growth

tumor cell

growth

sarcoma-180 cell

proliferation

HUVECs

growth

tumor cell

migration

human umbilical vein endothelial cell

metastasis

tumor cell

migration

HUVEC

metastasis

B16-F10 mouse melanoma cell

migration

HUVECs

activities

tumor

activities

angiogenesis

inhibiting

growth

inhibiting

metastasis

inhibited

migration

prevented

collagenolysis

suppressed

growth

suppressed

metastasis

effects

angiogenic

interference

proliferation

interference

migration

inhibition

collagenolysis

inhibition

effects

angiogenesis

inhibition

effects

growth

collagenolysis

purified

added

angiogenesis

injected

mice

induced

angiogenesis

TNF alpha

induced

TNF alpha

angiogenesis

caused

added

disrupted

induced

collagenase

collagenolysis

due

effects

interference

due

effects

inhibition