Production Of Pro-angiogenic Cytokines Research Articles

Immune response in pregnancy: An evolutionary challenge against external and internal stimuli

Immune response in pregnancy: An evolutionary challenge against external and internal stimuli

TNF-Like Weak Inducer of Apoptosis Promotes Angiogenesis, Thereby Exacerbating Cutaneous Psoriatic Disease

TNF-Like Weak Inducer of Apoptosis Promotes Angiogenesis, Thereby Exacerbating Cutaneous Psoriatic Disease

Inhibition of mTOR as Antiangiogenic Strategy in Multiple Myeloma

e240 Methods: Real-time RT PCR, Western blotting and immunofluorescence were performed to assess Notch and its ligands expression in MM endothelial cells (MMECs) and MGECs (ECs of patients with MGUS). Functional in vitro assay (Matrigel assay, adhesion, chemotaxis, wound-healing, cell proliferation, apoptosis, and ELISA) were conducted in MMECs and RPMI-8226 to study Notch inhibition effects after Notch2 silencing and -secretase treatment. Results: MMECs showed an higher expression and activation of Notch2 compared to MGECs and this higher activation in MMECs correlates whit an increased angiogenesis. Notch2 knock-down, through siRNA, reduced MMECs abilities and also their proliferation. Furthermore myeloma PCs line RPMI-8226 showed an over-expression of Notch ligands Jagged1, Jagged2 and DLL1 allowing mutual interactions between MMECs and RPMI8226 and Notch pathway activation, as demonstrated by co-colture experiments with or without transwell. Inhibition of Notch signaling using the -secretase inhibitor MK-0752, which prevents Notch cleavage, affected MMECs abilities in co-culture and alone. The block of Notch cleavage reduced ECs chemotaxis, migration, adhesion and furthermore the ability to form capillary structure on Matrigel . Moreover the treatment decreased the production of pro-angiogenic cytokines and of cell-cycle regulators in MMECs and in RPMI-8226 respectively. Conclusion: Our results show that Notch pathway is involved in MM progression supporting BM angiogenesis through an active interaction between myeloma PCs and MMECs. The inhibition of Notch signaling affected MMECs angiogenic abilities, highlighting the idea that this pathway could became a new target in MM therapy to counteract BM angiogenesis.

Rac1/Pak1/p38/MMP-2 Axis Regulates Angiogenesis in Ovarian Cancer.

Zoledronic acid is being increasingly recognized for its antitumor properties, but the underlying functions are not well understood. In this study, we hypothesized that zoledronic acid inhibits ovarian cancer angiogenesis preventing Rac1 activation. The biologic effects of zoledronic acid were examined using a series of in vitro [cell invasion, cytokine production, Rac1 activation, reverse-phase protein array, and in vivo (orthotopic mouse models)] experiments. There was significant inhibition of ovarian cancer (HeyA8-MDR and OVCAR-5) cell invasion as well as reduced production of proangiogenic cytokines in response to zoledronic acid treatment. Furthermore, zoledronic acid inactivated Rac1 and decreased the levels of Pak1/p38/matrix metalloproteinase-2 in ovarian cancer cells. In vivo, zoledronic acid reduced tumor growth, angiogenesis, and cell proliferation and inactivated Rac1 in both HeyA8-MDR and OVCAR-5 models. These in vivo antitumor effects were enhanced in both models when zoledronic acid was combined with nab-paclitaxel. Zoledronic acid has robust antitumor and antiangiogenic activity and merits further clinical development as ovarian cancer treatment.

Antitumor effect and antiangiogenic potential of the mTOR inhibitor temsirolimus against malignant pleural mesothelioma

The mTOR inhibitor temsirolimus has antitumor and antiangiogenic activity against several carcinomas, yet few reports document the efficacy of temsirolimus against malignant pleural mesothelioma (MPM). Therefore, we evaluated the efficacy of temsirolimus and the antiangiogenic effect of temsirolimus in the treatment of MPM. We examined the efficacy of temsirolimus alone and the efficacy of the combination of temsirolimus and cisplatin or pemetrexed against four MPM cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The effect of temsirolimus on the production of proangiogenic cytokines by MPM cell lines was examined by enzyme-linked immunosorbent assay (ELISA). Expression of mTOR and proangiogenic cytokines in clinical specimens from MPM patients was determined by immunohistochemistry. Temsirolimus inhibited cell viability and suppressed cell proliferation of all MPM cell lines. Combined treatment with temsirolimus and cisplatin inhibited the viability of all MPM cell lines more effectively than temsirolimus alone. Temsirolimus strongly inhibited the phosphorylation of p70s6k, a downstream molecule of mTOR, in all MPM cell lines and led to an increase in the levels of cleaved caspase-3 in the H226 and Y-meso14 cells. Temsirolimus also inhibited the production of vascular endothelial growth factor (VEGF) and platelet-derived growth factor-AA (PDGF-AA). Phosphorylated mTOR and high expression of VEGF and PDGF were detected in 2 and 3, respectively, out of the 5 MPM specimens. These results suggest that temsirolimus has activity against MPM cells by inhibition of cell proliferation and angiogenesis, and may be beneficial for a subset of MPM patients with high mTOR expression.

Abstract 4641: IL-16 in association with VEGFR-2 detect early tumor associated changes in the ovary.

Abstract Background: Ovarian cancer (OVCA) is a lethal malignancy of women with high rate of death among gynecological cancers. Due to the lack of an effective early detection test, OVCA in most cases are detected at late stage. Interleukin (IL)-16 is a cytokine involved in multiple immunopathobiological processes including inflammatory responses and production of proangiogenic cytokines. IL-16 is expressed by malignant epithelium and secreted in serum. Ovarian malignant transformation is followed by neovascularization and vascular endothelial growth factor receptor-2 (VEGFR-2) is a marker of tumor associated neovascularization (TAN). Thus, IL-16 and VEGFR-2 represent markers of OVCA at early stage may constitute an early detection test for OVCA. Objectives: The goal of this study was to determine whether IL-16 expression is associated with ovarian malignant transformation and to examine association between serum IL-16 levels and VEGFR-2 expression by TAN vessels during ovarian tumor development. Materials and methods: Normal (n =15) White Leghorn laying hens (3-4 years old) or hens with ovarian tumors (n=32) were selected by contrast enhanced transvaginal ultrasound scanning. Following serum collection, hens were euthanized, tissues were processed for routine histology, immunohistochemistry (IHC), protein and gene expression studies. Ovarian tumors were confirmed by gross morphology and microscopy. Expression of IL-16 by ovarian malignant epithelium and VEGFR-2 by ovarian TAN vessels were determined by IHC, immunoblotting and quantitative PCR. Serum IL-16 levels were determined by immunoassay. Correlation between serum IL-16 levels and frequencies of VEGFR-2 expressing microvessels were examined. Results: The intensity of IL-16 expression by normal ovarian epithelium was very weak (n=15). In contrast, the staining intensity of IL-16 by malignant epithelium increased significantly (p<0.001) in hens with early stage OVCA (n=15) and late stage OVCA (n=17). Compared with normal hens, serum levels of IL-16 were significantly higher in hens with early (n=15) and late stages (n=17) of OVCA (P< 0.0001). The expression of IL-16 protein and mRNA were stronger in malignant ovaries than normal ovaries. The population of VEGFR-2 expressing microvessels increased significantly (p<0.001) in hens with early stage OVCA than normal hens (p<0.001) and increased further in late stage OVCA. The increase in serum IL-16 was positively correlated with the population of VEGFR-2 expressing TAN vessels. Conclusion: IL-16 expression by ovarian malignant epithelium and its serum levels are associated with ovarian malignant transformation and correlated with VEGFR-2 expression. Thus IL-16 may be a useful serum marker and in combination with VEGFR-2 may detect OVCA at early stage. These findings will facilitate clinical studies to establish a non-invasive early OVCA detection test. Support: DOD Award # W81XWH-12-1-0460. Citation Format: Aparna Yellapa, Pincas Bitterman, Jacques S. Abramowicz, Janice M. Bahr, Sanjib Basu, Sameer Sharma, Animesh Barua. IL-16 in association with VEGFR-2 detect early tumor associated changes in the ovary. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4641. doi:10.1158/1538-7445.AM2013-4641

468 Circulating angiogenic cells plated on collagen IV matrices leads to enhanced endothelial cell formation and pro-angiogenic cytokine production

Circulating angiogenic cells (CACs) have been implicated in regenerating ischemic tissues following trauma such as myocardial infarction. Although controversial, evidence exists that a very small proportion of CACs may differentiate into endothelial cells (ECs) and integrate into developing vasculature. Collagen IV is a small, non-fibrillar collagen found in the basement membrane surrounding blood vessels and has been observed to be pro-neovasculogenic. We have previously demonstrated the benefit of injecting collagen-based matrices for revascularization of ischemic tissues.

E7080, a Multi–Tyrosine Kinase Inhibitor, Suppresses the Progression of Malignant Pleural Mesothelioma with Different Proangiogenic Cytokine Production Profiles

Malignant pleural mesothelioma (MPM) is a biologically heterogeneous malignant disease with a poor prognosis. We reported previously that the anti-vascular endothelial growth factor (VEGF) antibody, bevacizumab, effectively inhibited the progression of VEGF-high-producing (but not VEGF-low-producing) MPM cells in orthotopic implantation models, indicating the need for novel therapeutic strategies to improve the poor prognosis of this disease. Therefore, we focused on the multi-tyrosine kinase inhibitor E7080 and assessed its therapeutic efficacy against MPM cells with different proangiogenic cytokine production profiles. The efficacy of E7080 was assayed in orthotopic implantation of severe combined immunodeficient mouse models with three human MPM cell lines (MSTO-211H, NCI-H290, and Y-MESO-14). With regard to proangiogenic cytokine production profiles, MSTO-211H and Y-MESO-14 cells were MPM cells producing high levels of fibroblast growth factor-2 and VEGF, respectively. NCI-H290 cells produced low levels of fibroblast growth factor-2 and VEGF compared with the other two cell lines. E7080 potently suppressed the phosphorylation of VEGF receptor-2 and FGF receptor 1 and, thus, inhibited proliferation of endothelial cells, but not that of the MPM cell lines, in vitro. Orthotopically inoculated MSTO-211H cells produced only thoracic tumors, whereas NCI-H290 and Y-MESO-14 cells also developed pleural effusions. Treatment with E7080 potently inhibited the progression of these three MPM cell lines and markedly prolonged mouse survival, which was associated with decreased numbers of tumor-associated vessels and proliferating MPM cells in the tumor. These results strongly suggest broad-spectrum activity of E7080 against MPM with different proangiogenic cytokine production profiles in humans.

W1586 Requirement of ERK and Wnt/β-Catenin Signaling Pathways for G1-S Phase Transition of Non Immortalized Human Intestinal Epithelial Crypt Cells

nase-1 expression. Conclusion: Activation of CRHR1 increases angiogenesis, while activation of CRHR2 suppresses it. Additionally, activation of CRHRs in colonocytes modulates proangiogenic cytokine production. Thus, the CRH family peptides regulate intestinal angiogenesis In Vitro at both the endothelial and epithelial cell level, and this response may be important on the mechanism of CRH-mediated intestinal inflammation. Supported by a Young Clinical Scientific Award from “FAMRI” (E.I. and S.H.R.), NIDDK K01 DK079015” (S.H.R.) and PO-1 DK 33506 (C.P.)

Role of the Chemokine Receptor CXCR2 in Bleomycin-Induced Pulmonary Inflammation and Fibrosis

Pulmonary fibrosis is characterized by chronic inflammation and excessive collagen deposition. Neutrophils are thought to be involved in the pathogenesis of lung fibrosis. We hypothesized that CXCR2-mediated neutrophil recruitment is essential for the cascade of events leading to bleomycin-induced pulmonary fibrosis. CXCL1/KC was detected as early as 6 hours after bleomycin instillation and returned to basal levels after Day 8. Neutrophils were detected in bronchoalveolar lavage and interstitium from 12 hours and peaked at Day 8 after instillation. Treatment with the CXCR2 receptor antagonist, DF2162, reduced airway neutrophil transmigration but led to an increase of neutrophils in lung parenchyma. There was a significant reduction in IL-13, IL-10, CCL5/RANTES, and active transforming growth factor (TGF)-beta(1) levels, but not on IFN-gamma and total TGF-beta(1,) and enhanced granulocyte macrophage-colony-stimulating factor production in DF2162-treated animals. Notably, treatment with the CXCR2 antagonist led to an improvement of the lung pathology and reduced collagen deposition. Using a therapeutic schedule, DF2162 administered from Days 8 to 16 after bleomycin reduced pulmonary fibrosis and levels of active TGF-beta(1) and IL-13. DF2162 treatment reduced bleomycin-induced expression of von Willebrand Factor, a marker of angiogenesis, in the lung. In vitro, DF2162 reduced the angiogenic activity of IL-8 on human umbilical vein endothelial cells. In conclusion, we show that CXCR2 plays an important role in mediating fibrosis after bleomycin instillation. The compound blocks angiogenesis and the production of pro-angiogenic cytokines, and decreases IL-8-induced endothelial cell activation. An effect on neutrophils does not appear to account for the major effects of the blockade of CXCR2 in the system.

Mast Cells Contribute to Vasculogenic Mimicry in Multiple Myeloma

The angiogenic response is amplified during the induction phase of multiple myeloma (MM) and appears to exert a key role in the development of the disease [1]. Thus, inhibitors of angiogenesis have proven therapeutic potential in the treatment of patients with MM. Angiogenesis induced during the development of MM involves the direct production of proangiogenic cytokines by plasma cells within the marrow microenvironment. Mast cells (MCs) contribute to the the composition of the cellular components of the microenvironment in patients with MM, but their role in the pathophysiology of the disease is not clear. In this report, we used electron and confocal microscopy approaches to investigate the participation of MCs in the formation of the vessel wall in biopsy specimens from patients with MM. Results were compared to those obtained from the biopsy material of patients with a benign lesion, namely monoclonal gammopathy of undetermined significance (MGUS). Our results show that patients with MM exhibit typical tryptase-positive MCs, which interact physically with the endothelial cells (ECs) lining the vascular lumina, perhaps as a result of dysregulated vasculogenic development. This evidence highlights the importance of the stromal microenvironment during angiogenesis in the pathophysiology of MM and provides a novel perspective into the complex interplay between stromal and vascular components in the bone marrow microenvironment involved in the induction of hyervascularization in MM.

ROS production and angiogenic regulation by macrophages in response to heat therapy

Purpose: It has been well established that inadequate blood supply combined with high metabolic rates of oxygen consumption results in areas of low oxygen tension (<1%) within malignant tumours and that elevating tumour temperatures above 39°C results in significant improvement in tumour oxygenation. Macrophages play a dual role in tumour initiation and progression having both pro-tumour and anti-tumour effects. However, the response of macrophages to heat within a hypoxic environment has not yet been clearly defined.Methods: Raw 264.7 murine macrophages were incubated under normoxia and chronic hypoxia at temperatures ranging from 37–43°C. Under normoxia at 41°C, macrophages start to release significant levels of superoxide. The combination of heat with hypoxia constitutes an additional stimulus leading to increased respiratory burst of macrophages.Results: The high levels of superoxide were found to be associated with changes in macrophage production of pro-angiogenic cytokines. While hypoxia alone (37°C) increased levels of hypoxia inducible factor-1α (HIF-1α) in macrophages, the combination of hypoxia and mild hyperthermia (39–41°C) induced a strong reduction in HIF-1α expression. The HIF-regulated vascular endothelial growth factor (VEGF) decreased simultaneously, revealing that heat inhibits both HIF-1α stabilization and transcriptional activity.Conclusion: The data suggest that temperatures which are readily achievable in the clinic (39–41°C) might be optimal for maximizing hyperthermic response. At higher temperatures, these effects are reversed, thereby limiting the therapeutic benefits of more severe hyperthermic exposure.

Temporal Onset of Chronic Oxidative Stress and Hypoxia After Pulmonary Irradiation Is Associated with Profibrogenic and Proangiogenic Cytokine Production

Temporal Onset of Chronic Oxidative Stress and Hypoxia After Pulmonary Irradiation Is Associated with Profibrogenic and Proangiogenic Cytokine Production

Regulation of the pro-angiogenic microenvironment by carboxyamido-triazole.

Anti-angiogenic agents regulate tumor growth by inhibiting endothelial cell proliferation and invasion. Carboxyamido-triazole (CAI), an inhibitor of non-voltage-operated calcium entry and calcium influx-mediated pathways, has angiogenesis and invasion inhibitory activity. We hypothesized that CAI may express its anti-angiogenic effects through negative regulation of pro-angiogenic cytokine production and/or function. In vivo, orally administered CAI prevented A2058 human melanoma xenograft growth and concomitantly resulted in a marked reduction in circulating vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8). In vitro, A2058 cell secretion of VEGF was inhibited by CAI treatment under limiting micronutrient conditions that approximate the tumor microenvironment, media restriction, and acidification to pH 6.8 (P=0.0003 and P=0.0006, respectively). VEGF and HIF-1alpha message and protein were also reduced by CAI treatment. Oral CAI treatment reduced vascular ingrowth in vivo into VEGF-containing Matrigel plugs. Commensurate with those findings, human umbilical vein endothelial cell (HUVEC) migration towards VEGF was reduced below background by exposure to CAI in the migration chamber (P<0.0001). An 88% reduction in circulating IL-8 concentration was measured in CAI-treated animals. However, IL-8 protein secretion and gene expression were increased by CAI treatment in culture (P< or =0.01), where CAI caused a dose-dependent acidification of the culture milieu (P< or =0.005). This paradox suggests that IL-8 production in vitro may be more sensitive to ambient pH than cytosolic calcium. These observations suggest that CAI inhibition of tumor cell VEGF production and endothelial cell response to VEGF results in disruption of signaling between the tumor and its microenvironment, causing a net anti-angiogenic effect.

Exploiting the differential production of angiogenic factors within the tumor microenvironment in the design of a novel vascular-targeted gene therapy-based approach to the treatment of cancer

Purpose: The aim of this study is to explore a novel strategy through which the differential production of pro-angiogenic cytokines within the tumor microenvironment can be exploited as a means of selectively killing the vascular endothelial cells upon which the survival and growth of a tumor depend.Methods and Materials: Adenoviral vectors encoding a chimeric cell surface receptor composed of the extracellular domain of the vascular endothelial growth factor (VEGF) receptor Flk-1/KDR fused in frame to the membrane spanning and cytoplasmic domain of Fas were constructed and used to transduce primary human endothelial cells in vitro. The apoptotic response of these cells induced upon ligation of the chimeric receptor with VEGF was determined by measuring caspase-3 activation, AnnexinV-FITC binding, and the release of glucose-6-phosphate dehydrogenase.Results: The chimeric Flk-1/Fas protein is stable and expressed at high levels on the surface of adenovirally transduced cells. Upon the addition of exogenous VEGF, these cells undergo rapid apoptosis.Conclusions: Receptor/Fas chimeras that recognize and bind pro-angiogenic cytokines represent a novel means by which the signal transduction events normally triggered in vascular endothelial cells upon the binding of angiogenic cytokines may be redirected toward the induction of apoptotic cell death. It is proposed that these constructs will prove of value in the further development of safe and effective vascular-targeted gene therapy-based approaches to the treatment of cancer.

Production of Proangiogenic Cytokines During Thalidomide Treatment of Multiple Myeloma

Recently a growing number of studies have suggested the efficacy of thalidomide (THAL) in the treatment of relapsed or resistant multiple myeloma. Some of these studies indicate that the thalidomide antimyeloma effect is associated with decreased vessel density. Here we first present our experience with THAL treatment and then focus on the determination of the role of proangiogenic cytokines during THAL therapy. Thirty relapsing or resistant multiple myeloma (MM) patients were treated with THAL at a median dose of 400 mg/daily. Eighteen responded to THAL therapy and 12 were resistant or intolerant to THAL. We determined the plasma level of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) as the main biological parameters associated with tumour angiogenesis. In addition I1-6 and TNF α levels were also assayed. Assessment of peripheral blood (PB) and bone marrow (BM) cytokine levels were done before and during THAL treatment at weeks 4 and 8 of therapy. In the responder group VEGF, bFGF I1-6 and TNF α concentrations were significantly decreased after four weeks of therapy both in PB and BM. In the non-responder group no significant changes in bFGF and VEGF levels were observed. However, a significant increase in IL-6 and TNF concentrations was evident. We conclude that the significant decrease of VEGF, bFGF, I1-6 and TNF α concentrations reflected response to THAL therapy. Also it seems that VEGF is a better marker of response to treatment than bFGF.

Production of Proangiogenic Cytokines During Thalidomide Treatment of Multiple Myeloma

Production of Proangiogenic Cytokines During Thalidomide Treatment of Multiple Myeloma