Lower VEGF Research Articles

Bevacizumab and cetuximab with conventional chemotherapy reduced pancreatic tumor weight in mouse pancreatic cancer xenografts.

Pancreatic cancer remains the fourth leading cause of cancer-related death in the USA with a 5-year survival rate of 5%. The effects of epidermal growth factor receptor and vascular endothelial growth factor A blockade with chemotherapy on pancreatic tumor growth were examined. Mice bearing human PANC-1 cell xenografts were divided into three groups: T-CR (gemcitabine, cisplatin, and 5-fluorouracil), T-TR (cetuximab, bevacizumab, gemcitabine, cisplatin, and 5-fluorouracil), and vehicle control (T). The therapies were administered via intraperitoneal injections every 4days for seven cycles from 7weeks after cancer cell implantation. Mice treated with T-TR had significant reductions in tumor weight as compared to the control group (p<0.05). Although mice in the T-CR group experienced a significant reduction in body weight gain, serum albumin, and gastrocnemius muscle mass (p<0.05), no such reductions were observed in the T-TR group. Mice treated with T-TR had slightly increased CD11c+ DC and CD49b+ NK cell levels in the spleen (p<0.05) and significantly lower tumor VEGF expression (p<0.05). Tumor carcinoembryonic antigen expression was significantly reduced in both treatment groups (p<0.05). Thus, addition of bevacizumab and cetuximab to gemcitabine, cisplatin, and fluorouracil may represent an effective treatment option for pancreatic cancer that warrants further study.

Long-term safety and stability of angiogenesis induced by balanced single-vector co-expression of PDGF-BB and VEGF164 in skeletal muscle.

Therapeutic angiogenesis by growth factor delivery is an attractive treatment strategy for ischemic diseases, yet clinical efficacy has been elusive. The angiogenic master regulator VEGF-A can induce aberrant angiogenesis if expressed above a threshold level. Since VEGF remains localized in the matrix around expressing cells, homogeneous dose distribution in target tissues is required, which is challenging. We found that co-expression of the pericyte-recruiting factor PDGF-BB at a fixed ratio with VEGF from a single bicistronic vector ensured normal angiogenesis despite heterogeneous high VEGF levels. Taking advantage of a highly controlled gene delivery platform, based on monoclonal populations of transduced myoblasts, in which every cell stably produces the same amount of each factor, here we rigorously investigated a) the dose-dependent effects, and b) the long-term safety and stability of VEGF and PDGF-BB co-expression in skeletal muscle. PDGF-BB co-expression did not affect the normal angiogenesis by low and medium VEGF doses, but specifically prevented vascular tumors by high VEGF, yielding instead normal and mature capillary networks, accompanied by robust arteriole formation. Induced angiogenesis persisted unchanged up to 4 months, while no tumors appeared. Therefore, PDGF-BB co-expression is an attractive strategy to improve safety and efficacy of therapeutic angiogenesis by VEGF gene delivery.

Effect of combined consideration of distinct signatures of VEGF and soluble VEGFR2 on prognostic implication in gastric cancer.

56 Background: Anti-angiogenic strategy in gastric cancer (GC) has been highlighted again due to the recent success of ramucirumab and apatinib. Therefore, the comprehensive network of VEGF, soluble VEGF receptor-2 (sVEGFR2) and cytokines and other angiogenic factors (CAF) in GC and their prognostic impact would be of importance, although they have been poorly understood. We aimed to find out the CAF signature associated with VEGF and sVEGFR2, and to explore their prognostic implication in GC. Methods: We measured pretreatment serum levels of 52 CAFs, including VEGF and sVEGFR2, using multiplex bead immunoassays and ELISA, in 70 patients who were diagnosed with GC in Seoul National University Hospital, and treated with palliative chemotherapy. Linear regression analysis for correlating CAFs with VEGF and sVEGFR2, and survival analysis by log rank test and Cox regression analysis were performed. Results: The VEGF signature was shown to be associated with seven CAFs (interluekin [IL]-7, IL-12p70, IL-2Ra, IL-10, stem cell factor, Fibroblast growth factor-basic, IL-3). The sVEGFR2 signature was associated with IL-4 and platelet-derived growth factor beta, but VEGF and sVEGFR2 showed no association with each other. Patients with high VEGF had a tendency to have worse overall survival (OS) than those with low VEGF (11.2 months versus 16.7 months; P = 0.061). However, among patients with high-sVEGFR2, OS was not different according to VEGF (12.1 months, high-VEGF versus 15.1 months, low-VEGF; P = 0.546). Interestingly, the poor prognostic impact of high-VEGF was far significant in patients with low-sVEGFR2 (10.9 months versus 16.8 months; P = 0.036). With this perspective, VEGF/sVEGFR2 ratio was significantly correlated with worse OS in univariate as well as multivariate analysis (HR 1.78 [95% CI 1.08-2.94], P= 0.024). Conclusions: Based on the comprehensive network analysis of CAF, VEGF and sVEGFR2 had distinct CAF signatures in GC. Consideration of both VEGF and sVEGFR2 confers more accurate prognostic implication compared with VEGF alone in GC. Regarding the angiogenic aspect, VEGF/sVEGFR2 ratio is significantly correlated with survival outcome in GC.

PKCδ inhibition normalizes the wound-healing capacity of diabetic human fibroblasts.

Abnormal fibroblast function underlies poor wound healing in patients with diabetes; however, the mechanisms that impair wound healing are poorly defined. Here, we evaluated fibroblasts from individuals who had type 1 diabetes (T1D) for 50 years or more (Medalists, n = 26) and from age-matched controls (n = 7). Compared with those from controls, Medalist fibroblasts demonstrated a reduced migration response to insulin, lower VEGF expression, and less phosphorylated AKT (p-AKT), but not p-ERK, activation. Medalist fibroblasts were also functionally less effective at wound closure in nude mice. Activation of the δ isoform of protein kinase C (PKCδ) was increased in postmortem fibroblasts from Medalists, fibroblasts from living T1D subjects, biopsies of active wounds of living T1D subjects, and granulation tissues from mice with streptozotocin-induced diabetes. Diabetes-induced PKCD mRNA expression was related to a 2-fold increase in the mRNA half-life. Pharmacologic inhibition and siRNA-mediated knockdown of PKCδ or expression of a dominant-negative isoform restored insulin signaling of p-AKT and VEGF expression in vitro and improved wound healing in vivo. Additionally, increasing PKCδ expression in control fibroblasts produced the same abnormalities as those seen in Medalist fibroblasts. Our results indicate that persistent PKCδ elevation in fibroblasts from diabetic patients inhibits insulin signaling and function to impair wound healing and suggest PKCδ inhibition as a potential therapy to improve wound healing in diabetic patients.

Transcription of Inflammatory Cytokine TNFα is Upregulated in Retinal Angiogenesis under Hyperoxia

Background/Aims: Hypoxia induces angiogenesis while hyperoxia promotes vasoregression in the retina. We investigated herein the effect of prolonged hyperoxia on retinal angiogenesis and the underlying mechanism in an oxygen-induced retinopathy (OIR) model. Methods: Vascular morphology was quantified in whole-mount retina from the mice subjected to the conventional OIR model (c-OIR) or the OIR model with prolonged hyperoxia (p-OIR). Expressions of genes related to angiogenesis were determined by real-time PCR. Results: p-OIR retinas showed few intraretinal neovascular tufts at the border of avascular zones, lacking preretinal neovascularization, whereas c-OIR retinas had numerous preretinal neovascularizations. p-OIR retinas demonstrated outgrowth of capillaries in the deep layers despite persistent hyperoxia and possess a larger avascular zone compared with the c-OIR retinas. The capillaries in the p-OIR retinas were well-formed in contrast to those in the c-OIR retinas. p-OIR retinas expressed significantly higher TNFα (∼4 fold) than c-OIR retinas. The expression of vascular endothelial growth factor, Erythropoietin, Angiopoietin 1 and 2 remained unchanged. Conclusion: Our data demonstrate that TNFα transcription is increased in hyperoxia-promoted retinal angiogenesis, implicating it, in association with low VEGF levels, as a possible proponent in retinal angiogenesis under hyperoxia.

Supplementation of maternal omega-3 fatty acids to pregnancy induced hypertension Wistar rats improves IL10 and VEGF levels

ObjectivesOur recent study demonstrates the beneficial effect of a combined supplementation of vitamin B12, folic acid, and docosahexaenoic acid in reducing the severity of pregnancy induced hypertension (PIH). It is also known to be associated with angiogenic imbalance and inflammation. The current study examines whether the individual/combined supplementation of folic acid, vitamin B12 and omega-3 fatty acid during pregnancy can ameliorate the inflammatory markers and restore the angiogenic balance in a rat model of PIH. Materials and methodsThere were total of six groups, control and five treatment groups: PIH Induced; PIH+vitamin B12; PIH+folic acid; PIH+Omega-3 fatty acids and PIH+combined micronutrient supplementation (vitamin B12+folic acid+omega-3 fatty acids). Hypertension during pregnancy was induced using L- Nitroarginine methylester (L-NAME; 50mg/kg body weight/day). Dams were dissected at d20 of gestation and placental tissues were collected for further analysis. ResultsAnimals from the PIH induced group demonstrated lower (p<0.01 for both) IL-10 and VEGF levels as compared to control. However, PIH induction did not alter the protein levels of eNOS, IL-6, Flt and mRNA levels of VEGF and VEGFR-1/ Flt-1. Individual micronutrient supplementation of vitamin B12 and folate did not offer benefit. In contrast individual omega-3 fatty acid as well as combined micronutrient supplementation showed IL-10 and VEGF levels comparable to that of control. ConclusionOmega 3 fatty acid supplementation plays a key role in reducing inflammation in pregnancy induced hypertension.

VEGF dose regulates vascular stabilization through Semaphorin3A and the Neuropilin-1+ monocyte/TGF-β1 paracrine axis.

VEGF is widely investigated for therapeutic angiogenesis, but while short-term delivery is desirable for safety, it is insufficient for new vessel persistence, jeopardizing efficacy. Here, we investigated whether and how VEGF dose regulates nascent vessel stabilization, to identify novel therapeutic targets. Monoclonal populations of transduced myoblasts were used to homogeneously express specific VEGF doses in SCID mouse muscles. VEGF was abrogated after 10 and 17 days by Aflibercept treatment. Vascular stabilization was fastest with low VEGF, but delayed or prevented by higher doses, without affecting pericyte coverage. Rather, VEGF dose-dependently inhibited endothelial Semaphorin3A expression, thereby impairing recruitment of Neuropilin-1-expressing monocytes (NEM), TGF-β1 production and endothelial SMAD2/3 activation. TGF-β1 further initiated a feedback loop stimulating endothelial Semaphorin3A expression, thereby amplifying the stabilizing signals. Blocking experiments showed that NEM recruitment required endogenous Semaphorin3A and that TGF-β1 was necessary to start the Semaphorin3A/NEM axis. Conversely, Semaphorin3A treatment promoted NEM recruitment and vessel stabilization despite high VEGF doses or transient adenoviral delivery. Therefore, VEGF inhibits the endothelial Semaphorin3A/NEM/TGF-β1 paracrine axis and Semaphorin3A treatment accelerates stabilization of VEGF-induced angiogenesis.

VEGF-A levels in bevacizumab-treated breast cancer patients: a systematic review and meta-analysis.

Bevacizumab may improve outcomes of patients with breast cancer, but the absence of an established biomarker hampers patient selection and researchers´ ability to demonstrate a clear survival benefit. Its putative target, circulating VEGF-A, emerged as the main candidate and we sought to identify the relationship between VEGF-A levels and outcomes through systematic review. We searched electronic databases and meeting proceedings for randomized controlled trials (RCTs) comparing the addition of bevacizumab to standard chemotherapy for breast cancer. RCTs were included if outcomes were presented separately according to VEGF-A plasma levels. Random-effects model were applied to calculate the pooled hazard ratios for progression-free survival, event-free survival (EFS), comprising disease recurrence, progression or any-cause death, and overall survival (OS), with respective confidence intervals (95% CI). High and low VEGF-A levels subgroups followed each trial definition, and results were compared using the interaction test. Heterogeneity was calculated using χ (2) test (I (2)). Three trials enrolled a total of 3748 patients. 1713 patients had baseline VEGF-A levels in plasma available for assessment and were included. One trial added bevacizumab in the adjuvant setting (N=2591) and two on first-line metastatic disease with taxane-based therapy (N=1160) There was no interaction between VEGF-A levels and study setting (adjuvant vs. first line therapy). Bevacizumab improved PFS of patients with above median VEGF-A plasma levels (HR 0.56; 95% CI 0.43-0.73; P<0.001; I (2)=0%), but not of those with below median VEGF-A levels (HR 0.89; 95% CI 0.68-1.15; P=0.37; I (2)=0%), with relevant differences between these two groups, P-for interaction=0.02. The same happened with EFS (VEGF-A above median HR 0.62; 95% CI 0.39-0.79; P<0.001; I (2)=11%; below median HR 0.89; 95% CI 0.71-1.14; P=0.98; I (2)=17%; P-for interaction=0.03). OS data were not available. VEGF-A level is a reasonable candidate biomarker for bevacizumab in the treatment of breast cancer. Further studies have to confirm its surrogacy in overall survival and in other scenarios including other anti-angiogenic therapies.

Dickkopf-1 expression is down-regulated during the colorectal adenoma-carcinoma sequence and correlates with reduced microvessel density and VEGF expression.

Dickkopf-1 (Dkk1), an antagonist of the Wnt-β-catenin signalling pathway, has been reported to play a role in cancer progression. However, little is known about the role of Dkk1 during the colorectal adenoma-carcinoma sequence. This study aimed to elucidate the role of Dkk1 in tumorigenesis and angiogenesis in colorectal cancer. We examined Dkk1 expression immunohistochemically in 476 colorectal tissue samples, including 46 sets of matched specimens. Dkk1 expression was down-regulated during the colorectal adenoma-carcinoma sequence, both among the 476 samples and in the 46 sets of matched specimens. Dkk1 expression was correlated with decreased microvessel density (P < 0.05) and VEGF expression. In-vitro 3D coculture experiments showed that Dkk1 overexpression in HCT116 cells inhibited tube-like structure formation and down-regulated VEGF expression in human umbilical vein endothelial cells. Xenografts of Dkk1-overexpressing colorectal cancer cells were smaller, and showed lower microvessel density and VEGF expression levels, than those of control cells. This study is the first to show the roles of Dkk1 during the colorectal adenoma-carcinoma sequence, which may involve suppression of the tumorigenesis and angiogenesis of CRC. Dkk1 could therefore serve as a potential target for tumour therapy.

Risk factors for the Long-Term Efficacy, Recurrence, and Metastasis in Small Hepatocellular Carcinomas.

We tried to determine the risk factors for the long-term efficacy, recurrence, and metastasis of small hepatocellular carcinoma (HCC, diameter <5cm). One hundred sixty-eight small liver cancer patients received percutaneous cryoablation therapy by argon-helium superconducting surgery system under the ultrasound guidance. Clinical parameter and the efficacy were analyzed after follow-up. After cryoablation treatment, the median follow-up time for the 168 patients was 36 (7-41) months. Liver functions were impaired as indicated by increased alanine aminotransferase, total bilirubin, total protein, albumin, and prothrombin activity. The difference of VEGF expression in liver cancer and the surrounding tissue is significant. 1-, 2-, and 3-year overall survival were 92.9, 83.9, and 65.5%, respectively. Relapse-free survival was 76.8, 53.0, and 41.1%. Less tumor number, higher tumor differentiation, and low VEGF expression predict higher metastasis-free and relapse-free survival rate. Lower Child-Pugh classification is correlated with the higher overall survival after cryoablation. There was no statistical significance in in situ intrahepatic recurrence patients, but VEGF changes were statistically significant for metastasis in other parts of liver or extrahepatic metastasis. Tumor number, differentiation, VEGF expression, large vessel invasion, lymph node, and extrahepatic metastasis all affect the overall and relapse-free survival. VEGF expression can be a predictable factor for liver cancer recurrence and metastasis.

Notch expression and bevacizumab efficacy in colorectal cancer patients.

612 Background: Antiangiogenic therapies represent a well established additional treatment to standard chemotherapy, nevertheless no markers are available to suggest a successful outcome linked to the use of bevacizumab. To investigate potential mechanisms of resistance to angiogenesis inhibitor bevacizumab, Notch expression was correlated with response and survival in a series of bevacizumab treated advanced colorectal cancer (CRC) patients. Methods: Notch expression was evaluated by immunohistochemistry (IHC) on 65 primary CRC enrolled within 6 randomized clinical trials assessing first-line bevacizumab plus chemotherapy. Notch IHC was conducted using a polyclonal antibody to Cleaved Notch1 (Cell Signaling Technology, Danvers, MA). Notch expression was scored based on intensity of staining (0: none, 1+: weak, 2+: moderate; 3+: strong) and on percentage of immunostained cells. A control series of 21 advanced CRC treated with chemotherapy alone was also examined. Results: Notch positivity was localized to the cytoplasm or nucleus of malignant epithelial cells. In all, 11 of 61 (18%) evaluable primary tumours had a high Notch expression (IHC 3+). Six of the 11 cases (55%) with high Notch expression (IHC 3+) experienced progressive disease compared with 5 of 50 (10%) low Notch expression cases (IHC 0 1+ 2+) (p = 0.003). A high Notch expression also demonstrated an inferior median PFS (4.9 vs. 12.1 months; HR = 2.51; 95% CI, 0.96 to 6.58; p = 0.007) and OS (19.3 vs. 30.4 months; HR = 2.21; 95% CI, 0.79 to 6.15; p = 0.039) compared with low Notch expression cases. When the groups were further analyzed considering VEGF expression, the best outcome was found in low Notch (IHC 0 1 +) and high VEGF expressing tumors (IHC 2+ 3+) (response rate 9 of 11, 82 % vs. 1 of 5, 20%, in patients with high levels of Notch and VEGF expression, respectively). No correlation was found between Notch expression and clinical response in the series of patients treated with chemotherapy without bevacizumab. Conclusions: Notwithstanding the limited power of the present analysis, these data seem to suggest that low Notch expression might be a marker for successful bevacizumab treatment.

Short Pulse Laser Induces Less Inflammatory Cytokines in the Murine Retina after Laser Photocoagulation

Aims: The purpose of this study was to evaluate the effect of pulse duration on the expression of inflammatory cytokines in the murine retina after laser photocoagulation treatment with a PASCAL® pattern scan laser photocoagulator and conventional laser treatment. Methods: Retinal scatter laser photocoagulation was performed on C57BL/6J mice using a short pulse (10 ms) with a PASCAL laser or conventional settings (100 ms) with a multicolor laser. Eyes were enucleated before treatment (control) and 1 day, 3 days and 7 days after treatment. The levels of inflammatory cytokines (i.e., VEGF, MCP-1, RANTES and IL-6) in the retina/choroid were quantified by an ELISA. The expression patterns of VEGF and macrophages (i.e., F4/80) in the retina/choroid were evaluated by immunohistochemistry. Results: The levels of RANTES, IL-6 and MCP-1 after PASCAL and conventional laser treatments were significantly elevated compared with controls (p < 0.05). Conventional laser treatment, but not PASCAL treatment, resulted in the up-regulation of VEGF. RANTES and IL-6 levels on day 1 and MCP-1 levels on day 3 in the sensory retina were also significantly up-regulated with conventional laser treatment compared with PASCAL treatment (p < 0.05). Immunohistochemical analysis showed that PASCAL treatment was associated with lower VEGF and F4/80 expression levels compared with conventional laser treatment. Conclusions: Our data suggested that the short pulse duration induced fewer inflammatory cytokines in the sensory retina compared with the conventional pulse duration. Short pulse laser photocoagulation with the PASCAL may prevent macular edema after panretinal photocoagulation.

ME-03 * BONE MARROW DERIVED MICROGLIA AND THEIR FUNCTION WITHIN TUMOR MICROENVIRONMENT

We have previously demonstrated that BMDC are recruited to Glioblastoma Multiforme (GBM), in a highly tumor-growth and region-dependent pattern, demonstrating that the majority of BMDCs differentiate into inflammatory cells, MAC3 + , and CD11b+ IBA1+ microglia-like cells. The majority of microglia seen in GBM microenvironment are derived from the BMDC rather than being resident brain microglia. The role of microglial cells in GBMs remains unknown, with evidence supporting both an anti- and pro- tumorgenic function. In this study we aimed to understand the contribution of microglia to GBM growth and tumor vascularity. Chimeric mice with reconstituted green-fluorescent bone marrow were used to create intracranial GBM xenografts, by implanting red fluorescent glioma stem cells or U87 into the frontal lobe. Using a known inhibitor of microglial activation, minocycline, we show that treatment with this agent increased tumor cell growth and invasiveness and reduced animal survival compared to controls. We show that Minocycline decreases activation of microglia, inhibits the phagocytic activity of microglia in GBMs and decreases the perivascular localization of microglia. The mRNA expression profile of microglia isolated from the GBM microenvironment showed, through Ingenuity Pathway Analysis (IPA), a loss of engulfment and migration of cells and phaocytotic pathways in BM-derived microglia, additionally we see a reorganization of cytoskeletal pathways leading to an increase in the migration and invasiveness of GBM cells following treatment with Minocycline. We also have preliminary data to suggest that this property of Minocycline is dependent on VEGF levels, where in the context of low VEGF Minocycline does not provide a significant anti-angiogenic role. We demonstrate that microglia in GBM microenvironment, are primarily recruited and differentiated from the BM and not resident, and that these cells have an anti-tumorgenic functions, and inhibition of the activated microglia by Minocycline, results in a more invasive phenotype of GBM cells.

PO-0432 Serum Vascular Endothelial Factor (vegf) In Preterm Infants Predicts Chorioamnionitis But Not Abnormal Neuroimaging

Background Brain injury and inflammation are common mechanisms of disease in preterm infants. VEGF is an endothelial growth factor which has been shown to be neuro-protective in adult stroke. Infants less than 32 weeks’ gestation who suffer from respiratory distress syndrome (RDS) have been shown to have significantly lower VEGF levels than those without RDS. However little is known about the mechanisms of VEGF in the premature brain. Aim We aimed to investigate the relationship between brain injury and chorioamnionitis and serum VEGF levels in preterm infants. Methods Preterm infants Results Preterm infants (n = 86) had a mean gestation of 28+4 ± 0+2 weeks and Birth weight of 1165 ± 326 kg. Infants (n = 29) with abnormal cranial ultrasounds (n = 9) had a decreased mean VEGF level (227 ± 322 pg/mL versus normal 324 ± 528 pg/mL). Serum VEGF levels on Day 1 were significantly associated with histological chorioamnionitis (p = 0.0006). The mean serum VEGF levels were 116 ± 182 pg/mL in those without chorioamnionitis compared to 735 ± 683 pg/mL in those with chorioamnionitis. Conclusion Elevated serum VEGF is associated with choriomanionitis in preterm infants.

Abstract 5449: FP-1039/GSK3052230, an FGF ligand trap, enhances VEGF antagonist therapy in preclinical models of RCC and HCC

Abstract An increasing body of evidence has implicated FGF2 as one of the drivers of resistance to various inhibitors of VEGF-mediated angiogenesis. This resistance may play a role as a key limitation to the efficacy of therapies targeted at VEGF and its receptors. We investigated the potential for FP-1039/GSK3052230, a ligand trap that sequesters FGFs and inhibits their signaling, to enhance the activity of VEGF antagonist therapies in certain preclinical models of renal cell (RCC) and hepatocellular (HCC) carcinomas. First, we examined whether FP-1039/GSK3052230 has single agent efficacy against human RCC and HCC xenografts that express relatively high levels of FGF2, a profile that would mimic FGF2-driven resistance to VEGF therapy. We determined that this expression profile represents 34% of clear cell RCC (ccRCC) and 31% of HCC patients, based on the cancer genome atlas (TCGA) data. Human ccRCC xenografts with high FGF2 expression and low VEGFA expression demonstrated a significant inhibition in tumor growth when treated with FP-1039/GSK3052230 alone (TGI: 39-81%). In addition, we show that the high FGF2 expression profile is similarly predictive for the anti-tumor response of a human HCC model to single-agent FP-1039/GSK3052230 (TGI: 31-55%). In contrast, RCC models with low FGF2 expression, representing 66% of all ccRCC in the TCGA, are relatively insensitive to FP-1039/GSK3052230 as a single-agent. However, combination therapy of FP-1039/GSK3052230 with pazopanib in these tumors is significantly more effective than either agent alone. FP-1039/GSK3052230 not only slows tumor growth, but can induce ∼25% tumor regression when administered to mice bearing ccRCC xenografts that have become resistant to pazopanib. Together, our data demonstrate that FP-1039/GSK3052230 may be an effective therapy against RCC and HCC, both as a single agent in disease driven by FGF2 and in combination with VEGF antagonist therapies that represent the current standards of care for advanced disease. Citation Format: David I. Bellovin, Servando Palencia, Kevin Hestir, Ernestine Lee, M. Phillip DeYoung, Thomas Brennan, Gerrit Los, Kevin Baker. FP-1039/GSK3052230, an FGF ligand trap, enhances VEGF antagonist therapy in preclinical models of RCC and HCC. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5449. doi:10.1158/1538-7445.AM2014-5449

Assessment of uterine, subendometrial blood flows and endometrial gland vascular endothelial growth factor (EG-VEGF) in women with unexplained infertility

Abstract Objective To asses uterine and subendometrial blood flows as well as the endometrial gland vascular endothelial growth factor (EG-VEGF) as markers of endometrial angiogenesis and receptivity in women with unexplained infertility. Methods The study included two groups of women: The Unexplained Infertility Group, included fifty women presented to the infertility outpatient clinic with unexplained primary infertility; and the Fertile Group, included fifty fertile parous women presented to the family planning clinic seeking contraception. On day 6 after detection of urinary LH surge, transvaginal ultrasound scan (TVS) was done for measuring endometrial thickness, uterine artery and subendometrial Doppler velocimetry indices. On the same day of TVS, endometrial samples were taken using office suction sampler for immunohistochemical detection of the anti-vascular endothelial growth factor antibody. Results Women of the Unexplained Infertility Group had lower VEGF score, thinner endometrial thickness, higher subendometrial flow resistance index (RI) and lower subendometrial flow pulsatility index (PI). Conclusion Subendometrial, but not the uterine, blood flow and the EG-VEGF score seem to be significantly lower during the mid-luteal phase in women with unexplained infertility. This may suggest significantly poorer angiogenesis during the assumed peri-implantation period.

Benzaldehyde suppresses murine allergic asthma and rhinitis.

To evaluate the antiallergic effects of oral benzaldehyde in a murine model of allergic asthma and rhinitis, we divided 20 female BALB/c mice aged 8-10 weeks into nonallergic (intraperitoneally sensitized and intranasally challenged to normal saline), allergic (intraperitoneally sensitized and intranasally challenged to ovalbumin), and 200- and 400-mg/kg benzaldehyde (allergic but treated) groups. The number of nose-scratching events in 10 min, levels of total and ovalbumin-specific IgE in serum, differential counts of inflammatory cells in bronchoalveolar lavage (BAL) fluid, titers of Th2 cytokines (IL-4, IL-5, IL-13) in BAL fluid, histopathologic findings of lung and nasal tissues, and expressions of proteins involved in apoptosis (Bcl-2, Bax, caspase-3), inflammation (COX-2), antioxidation (extracellular SOD, HO-1), and hypoxia (HIF-1α, VEGF) in lung tissue were evaluated. The treated mice had significantly fewer nose-scratching events, less inflammatory cell infiltration in lung and nasal tissues, and lower HIF-1α and VEGF expressions in lung tissue than the allergic group. The number of eosinophils and neutrophils and Th2 cytokine titers in BAL fluid significantly decreased after the treatment (P<0.05). These results imply that oral benzaldehyde exerts antiallergic effects in murine allergic asthma and rhinitis, possibly through inhibition of HIF-1α and VEGF.

Tumor VEGF expression correlates with tumor stage and identifies prognostically different groups in patients with clear cell renal cell carcinoma

ObjectivesVascular endothelial growth factor (VEGF) is a potent inducer of tumor angiogenesis and represents the key element in the pathogenesis of clear cell renal cell carcinoma (ccRCC). The aim of this study was to investigate the use of tumor VEGF expression as a parameter to identify tumor stage and prognostically different patient groups. Methods and materialsWe retrospectively collected clinical data of 137 patients treated with partial or radical nephrectomy at our institutions for organ-confined, locally advanced, and metastatic ccRCCs between 1984 and 2013. Tumor cell VEGF immunohistochemical expression was compared with pathological and clinical features including age, sex, tumor stage, and Fuhrman grade. Comparison of VEGF expression levels between tumor stages was performed via Kruskal-Wallis nonparametric test. Survival analysis was conducted via Kaplan-Meier product-limit method, and Mantel-Haenszel log-rank test was employed to compare survival among groups. ResultsMedian age at diagnosis was 61 years (range: 33–85y). Tumor stage was pT1N0M0 in 67 patients (49%), pT2N0M0 in 5 (4%), and pT3N0M0 in 25 (18%), while 40 patients (29%) had metastatic tumors at diagnosis. Fuhrman nuclear grade was G1 in 22 patients (16%), G2 in 60 (44%), G3 in 33 (24%), G4 in 13 patients (9%), and unknown in 9 patients. Tumor VEGF was differentially expressed among different stages (P<0.001) and in low (G1–2) and high (G3–4) Fuhrman grade tumors (P<0.001). No significant differences were found when stratifying by sex (P = 0.06) or age (P = 0.29). Median overall survival (OS) from partial or radical nephrectomy was 161 months (range: 1–366). We observed a significantly longer OS in patients with low (<25%) vs. high (>25%) VEGF expression levels (median OS 206 vs. 65mo, P<0.001). ConclusionsOur data show that tumor cell VEGF expression is significantly associated with tumor stage and Fuhrman grade and is able to predict patient outcome, suggesting a potential use of this parameter in identifying prognostically different patients with ccRCC.

Serum level and immunohistochemical expression of vascular endothelial growth factor for the prediction of postoperative recurrence in renal cell carcinoma.

BackgroundVascular endothelial growth factor (VEGF) plays a major role in angiogenesis. One of the functions of VEGF is to regulate neovascularization in clear cell renal cell carcinoma (CCRCC). The objective of our study was to examine whether before nephrectomy serum levels of VEGF or expression of VEGF using immunohistochemistry (IHC) could predict postoperative recurrence in nonmetastatic CCRCC.ResultsTwelve patients (14.5%) had recurrence during a mean follow-up of 52.6 ± 31.2 months. The serum VEGF level was significantly higher in patients with recurrence than in those without recurrence (P = 0.038). High serum VEGF levels were above 416 pg/mL; this value was chosen based on a receiver operating characteristic analysis. The recurrence-free survival rate in patients with a high serum VEGF level was significantly lower than in those with a low serum VEGF level (P = 0.003). In total, tumors from 26 patients (31.3%) showed overexpression of VEGF using IHC. The recurrence-free survival rate in the IHC-positive group was significantly lower than that in the IHC-negative group (P = 0.044). Multivariate analysis indicated that preoperative serum VEGF levels (P = 0.013) and female gender (P = 0.004) were independent predictors of postoperative recurrence in nonmetastatic CCRCC.ConclusionsPreoperative serum VEGF levels is a useful predictor compared with IHC analysis of VEGF of postoperative recurrence in nonmetastatic CCRCC.

A Correlative Biomarker Analysis of the Combination of Bevacizumab and Carboplatin-Based Chemotherapy for Advanced Nonsquamous Non–Small-Cell Lung Cancer: Results of the Phase II Randomized ABIGAIL Study (BO21015)

Avastin Biomarkers In lunG And 3D Innovative anaLysis (ABIGAIL), which is a phase II, open-label, randomized study, investigated correlations between biomarkers and best overall response to bevacizumab plus platinum-doublet chemotherapy for patients with advanced/recurrent non-small-cell lung cancer. Patients received bevacizumab (7.5 or 15 mg/kg, 3-weekly until disease progression/unacceptable toxicity) plus carboplatin/gemcitabine or carboplatin/paclitaxel (maximum six cycles). Plasma samples (baseline/throughout treatment) were analyzed for vascular endothelial growth factor (VEGF)-A (baseline only), VEGF receptors (VEGFR-1/VEGFR-2), basic fibroblast growth factor, E-selectin, intercellular adhesion molecule-1, and placental growth factor (baseline only). Tumor samples (primary specimen) were analyzed for VEGF-A, VEGFR-1/VEGFR-2, neuropilin (NRP), and CD31. Response was evaluated at baseline and every 6 weeks (Response Evaluation Criteria in Solid Tumors). Patients were randomized to receive chemotherapy plus 7.5 mg/kg (n =154) or 15 mg/kg (n =149) bevacizumab. For the primary analysis, none of the baseline plasma biomarkers correlated with best overall response. Exploratory analyses showed that low VEGF-A levels were associated with longer progression-free survival (7.4 versus 6.1 months; hazard ratio, 1.57; 95% confidence intervals, 1.17 to 2.09; p = 0.002) and overall survival (19.8 versus 11.1 months; hazard ratio, 1.57; 95% confidence interval, 1.15-2.13; p = 0.004) compared with these in high baseline plasma VEGF-A levels. No plasma biomarkers changed significantly over time. No significant correlations were observed between tumor biomarkers and clinical outcomes. No new safety signals were observed. Baseline and/or dynamic changes in plasma basic fibroblast growth factor, E-selectin, intercellular adhesion molecule-1, placental growth factor, VEGFR-1 and VEGFR-2, and tumor biomarkers did not correlate statistically with treatment outcomes for bevacizumab plus chemotherapy. Only baseline plasma VEGF-A was significantly correlated with progression-free survival/overall survival.