sVEGFR-1 Levels Research Articles

Vascular endothelial growth factor and soluble vascular endothelial growth factor receptor-1 in patients with end-stage renal disease

Objectives:To show the levels of vascular endothelial growth factor (VEGF), soluble vascular endothelial growth factor receptor-1 (sVEGFR-1) in patients with end-stage renal disease (ESRD) and to show the associations with clinical findings such as demographic features, laboratory findings, comorbidities, and medications.Methods:A total of 73 people, consisting of patients with ESRD (n=38) and healthy subjects (n=35) in Gulhane Education and Research Hospital, Ankara, Turkey, were included in this cross-sectional study between the years 2011 and 2013. Blood samples were obtained and plasma VEGF, sVEGFR-1 analyzes were performed.Results:The VEGF level of ESRD group was not significantly higher (0.280±0.264) than the control group (0.321±0.210) (p=0.475). The sVEGFR-1 level of ESRD group was significantly higher (0.217±0.135) than the control group (0.068±0.047) (p<0.001). The correlation between VEGF and sVEGFR-1 was significant and negative (r=-0.246, p=0.036). Average VEGF level of ESRD patients using recombinant human erythropoietin (rhEPO) was significantly higher (0.567±0.28) than the ESRD patients not using rhEPO (0.246±0.24) (p=0.025).Conclusion:Our study is the first showing the significance of sVEGFR-1 in ESRD patients, and associations with comorbidities, medications. Especially our finding of rhEPO and VEGF may illuminate a reasonable positive effect of rhEPO on angiogenesis. Soluble vascular endothelial growth factor receptor-1 and VEGF may be important markers in the pathophysiology of ESRD.

Relationship between circulating vascular endothelial growth factor and its soluble receptor in patients with hemorrhagic fever with renal syndrome

Hemorrhagic fever with renal syndrome (HFRS) is characterized by endothelial dysfunction with capillary leakage without obvious cytopathology in the capillary endothelium. The aim of the study was to analyze the kinetics of vascular endothelial growth factor (VEGF) and its soluble receptor (sVEGFR-2) in HFRS patients infected with Dobrava (DOBV) or Puumala virus (PUUV). VEGF and sVEGFR-2 levels were measured in daily plasma and urine samples of 73 patients with HFRS (58 with PUUV, 15 with DOBV) and evaluated in relation to clinical and laboratory variables. In comparison with the healthy controls, initial samples (obtained in the first week of illness) from patients with HFRS had higher plasma and urine VEGF levels, whereas sVEGFR-2 levels were lower in plasma but higher in urine. VEGF levels did not differ in relation to hantavirus species, viral load, or the severity of HFRS. The comparison of VEGF dynamics in plasma and urine showed the pronounced secretion of VEGF in urine. Significant correlations were found between daily VEGF/sVEGFR-2 levels and platelet counts, as well as with diuresis: the correlations were positive for plasma VEGF/sVEGFR-2 levels and negative for urine levels. In addition, patients with hemorrhagic manifestations had very high plasma and urine VEGF, together with high urine sVEGFR-2. Measuring the local secretion of sVEGFR-2 in urine might be a useful biomarker for identifying HFRS patients who will progress to severe disease.

Angiogenic and inflammatory biomarkers for screening and follow-up in patients with pulmonary arterial hypertension

Objective: To identify circulating angiogenic and inflammatory biomarkers with potential in screening for pulmonary arterial hypertension (PAH) in systemic sclerosis (SSc), and in early diagnosis and determination of treatment response in PAH.Method: Plasma samples were taken at the time of PAH diagnosis and at treatment follow-up after a median (interquartile range) of 4 months (3–9.8 months) in idiopathic (n = 9) and SSc-associated PAH (n = 11). In patients with SSc-associated PAH, plasma samples had also been gathered a median of 2 years (0.8–3 years) before PAH diagnosis (n = 10). Additional plasma samples were retrieved at two time-points separated by a median of 12 years (10–13 years) from SSc patients who did not develop PAH (n = 10) and from controls (n = 8). Angiogenic and inflammatory biomarkers were analysed by multiplex immunoassays.Results: Plasma levels of placenta growth factor (PlGF), soluble vascular endothelial growth factor receptor-1 (sVEGFR-1), and tumour necrosis factor-α (TNF-α) were higher (p < 0.05) in SSc patients who later developed PAH than in those who did not. Plasma vascular endothelial growth factor (VEGF)-D increased (p < 0.05) in SSc patients as PAH developed. Plasma levels of PlGF, VEGF-A, VEGF-D, sVEGFR-1, interleukin-6, and TNF-α were higher (p < 0.05) in PAH than controls. There were no significant differences in circulating biomarkers between idiopathic and SSc-associated PAH. Plasma sVEGFR-1 decreased (p < 0.05) after initiating PAH-targeted treatments.Conclusions: Plasma levels of PlGF, sVEGFR-1, TNF-α, and VEGF-D have potential in screening for SSc-associated PAH. Plasma sVEGFR-1 may be a biomarker of treatment response.

Circulating angiogenesis-related markers as predictors of benefit from regorafenib in metastatic colorectal cancer (mCRC) patients (pts).

675 Background: Regorafenib is a treatment option for refractory mCRC pts. Considering its limited clinical benefit and the palliative setting, patients’ selection is essential to optimize the cost-effectiveness ratio. We aimed to describe the modulation of selected circulating angiogenic factors by regorafenib and to investigate their correlation with clinical outcome. Methods: IL-8, Ang-2, PDGF, bFGF, Tie-2, sVEGFR2, sVEGFR3, PlGF, VEGF-A, VEGF-B were assessed by ELISA on plasma samples collected at baseline (d1), after 15 days of treatment (d15), at the best RECIST response, at disease progression (PD) in mCRC pts treated with regorafenib, as per indication. Comparisons among concentrations of each marker at different timepoints were performed by Wilcoxon test. Markers showing significant changes were analyzed for correlation with outcome. Results: 105 pts were included. Median PFS and OS were 2.1 and 7.0 months (mos), respectively. As compared to d1, IL-8 and Ang-2 levels increased at PD. An early decrease at d15 was observed for PDGF, Tie-2, sVEGFR2 and sVEGFR3 levels, followed by an increase at PD. Conversely, PlGF levels increased at d15 and then decreased at PD. Baseline levels of Ang-2 and Tie-2 below the median value were associated with longer PFS (HR:0.61 [0.37-0.83] p = 0.006; HR:0.69 [0.43-0.95] p = 0.04, respectively) and OS (HR:0.45 [0.26-0.60] p &lt; 0.0001; HR:0.68 [0.44-0.98] p = 0.04, respectively). With regard to Ang-2, 40 (45%) out of 89 pts with available plasma samples at d15 showed increased levels at d15 as compared to d1. Among them, 21 (53%) achieved disease control, as compared to 14 out of 49 (29%) pts with Ang-2 decreased levels (p = 0.03). Median PFS of pts with increased and decreased Ang-2 levels were 3.1 and 1.8 mos, respectively (HR:0.57 [0.33-0.78] p = 0.004). Conclusions: A dynamic modulation of plasma angiogenic factors occurs during the treatment with regorafenib. Low baseline Ang-2 and Tie-2 levels seem to be associated with good prognosis. The early modulation of Ang-2 levels may predict benefit from regorafenib. These results need validation in independent series.

A phase 2 and biomarker study of sorafenib combined with FOLFOX in patients with advanced hepatocellular carcinoma (HCC).

270 Background: Sorafenib is the standard first line treatment for advanced HCC and showed a median time to progression (TTP) of 5.5 months and an overall response rate (ORR) of 2% in the phase III SHARP trial. FOLFOX has shown modest activity in HCC with a progression free survival (PFS) of 2.9 months and ORR of 8% in a phase III trial. In this single-arm, multicenter phase 2 and biomarker study, sorafenib plus FOLFOX was evaluated in the first line treatment of advanced HCC. Methods: Patients with histologically proven advanced HCC, Child Pugh A liver function, and no prior systemic therapies received sorafenib 400mg orally twice daily during a 2-week lead-in, followed by concurrent modified FOLFOX (5-FUCI 1200mg/m2/day for 46 hours and LV 400mg/m2 bolus, Oxaliplatin 85mg/m2) on day 1 and 15 of each 28-day cycle. The primary endpoint was TTP, calculated from date of study entry to date of radiological or clinical disease progression. Serial plasma anti-angiogenic and anti-inflammatory biomarkers were evaluated. Results: The study enrolled 40 patients with advanced HCC: median age, 65 years; male 85%; Child Pugh A5, 70%; BCLC stage C, 95%; HCC etiology, HCV 40%, HBV 13%, alcohol 13%. Grade 3/4 adverse events were notable for AST (23%), ALT (15%), bilirubin (10%), diarrhea (10%), anemia (10%), hypertension (5%), hand-foot syndrome (5%), and thrombocytopenia (5%). Dose reductions for sorafenib and FOLFOX were done in 73% and 65% of patients, respectively. The median TTP was 8.8 months (95%CI, 6.5-11.2). The ORR was 18%, and the stable disease rate was 55%. Among 36 patients with a baseline AFP ≥ 5 ng/mL, 10 (28%) had a ≥ 50% drop in AFP. Low baseline plasma levels of sVEGFR1, VEGF-C, and bFGF and high levels of s-cMET and IL-12 tended to associate with longer TTP (p &lt; 0.10). Decreased s-cMET at day 15 and decreased s-cMET and IL-2 at day 43 were associated with longer TTP (p &lt; 0.05). Conclusions: Sorafenib+FOLFOX demonstrated encouraging clinical efficacy with moderate toxicity in the first line treatment of advanced HCC. Initial biomarker evaluation suggested a correlation between TTP and baseline angiogenic markers as well as changes in IL-2 and s-cMET. Complete biomarker analysis will be presented at the meeting. Clinical trial information: NCT01775501.

VEGFR2 alteration in Alzheimer\u2019s disease

Alzheimer’s disease (AD) is a common disorder of progressive cognitive decline among elderly subjects. Angiogenesis-related factors including vascular endothelial growth factor (VEGF) might be involved in the pathogenesis of AD. Soluble form of the VEGF receptor is likely to be an intrinsic negative counterpart of VEGF. We measured the plasma levels of VEGF and its two soluble receptors (sVEGFR1 and sVEGFR2) in 120 control subjects, 75 patients with mild cognitive impairment, and 76 patients with AD using ELISA. Plasma levels of VEGF in patients with AD were higher than those in healthy control subjects. However, plasma levels of sVEGFR1 and sVEGFR2 were lower in patients with AD than in healthy control subjects. Levels of VEGFR2 mRNA were significantly decreased in human umbilical vein endothelial cells after amyloid-beta treatment. Further, protein levels of VEGFR2 were also decreased in the brains of AD model mice. In addition, we show that the expression of sVEGFR2 and VEGFR2 was also decreased by the transfection with the Notch intracellular domain. These results indicate that the alterations of VEGF and its two receptors levels might be associated with those at risk for Alzheimer’s disease.

Inducible knockout of Syncytin-A gene leads to an extensive placental vasculature deficiency, implications for preeclampsia

Syncytin-1, a human endogenous retroviral envelope gene (HERVW1), is specifically expressed in placental trophoblasts and mediates the formation of syncytiotrophoblasts through a fusogenic activity. Syncytin-1 expression deficiency has been repeatedly observed in preeclamptic/IUGR placentas. Previous gene knockout studies indicated that in mice, complete syncytin-A null mouse embryos died in utero between 11.5 and 13.5days of gestation. However, the complete knockout model could not fully recapitulate the mid- to third-trimester, time-specific syncytin-1 deficiency in preeclampsia patients. To construct a preeclampsia model and to better investigate the function of syncytin in placental development, we created a mouse inducible knockout model of syncytin-1A gene. It was found that the disruption of syncytin-A at E11.5 to E17.5 is associated with significant morphological changes in placentas and fetuses. Moreover, syncytin-A disruption led to extensive vasculature abnormalities in the labyrinth, with irregular distribution and reduced number of fetal microvessels. Moreover, Syncytin-A knockout affected neovascularization-related gene expression in labyrinth and the maternal plasma level of sVEGFR-1, and a dramatic increase of sFlt-1/PlGF ratio. These findings indicate that syncytin-A may be involved in the placenta angiogenesis and potentially, the development of preeclampsia. The new model could be a useful tool for studying the pathogenesis and management of preeclampsia.

Abstract 2734: Impact of circulating and tissue biomarkers in patients with metastatic colorectal cancer treated with first-line FOLFOX-aflibercept therapy. Results of the GERCOR VELVET Phase II Study

Abstract Background: The combination of aflibercept to OPTIMOX (VELVET study) was evaluated in patients with previously untreated advanced colorectal cancer. A biomarker program was set-up to explore the expression of several tissue biomarkers at baseline and circulating biomarkers upon treatment cycles to identify the best monitoring biomarkers of the treatment strategy. Methods: VELVET was a prospective, single arm phase II trial. Patient’s plasma samples were collected at baseline, and during induction therapy at day 1 of the first 6 cycles. Circulating biomarkers were analyzed using multiplexing immunoassays (31 biomarkers, 3 panels). Assays were conducted on a Biorad Bioplex platform, expect for PlGF, VEGF-A and Neuropillin-1 which were assessed using ELISA assays. Tissue biomarker were analyzed at baseline by IHC, using a BOND-Max autostainer (Leica biosystems). EGFR mutation status were analyzed using next generation sequencing. Results: Among 49 patients included in the VELVET study from May 2013 to May 2014, 44 (90%) patients were evaluable for circulating biomarkers expression. The proportion of patients with tumor response (CR or PR) was higher in patients with high baseline levels of sVEGFR2, sEGFR, G-CSF, Prolactin and low baseline levels of VEGFA and MIF. Progression-free survival (PFS) was higher in patients with low baseline levels of Osteopontin (HR: 0.53; P=0.045) and in patients with high baseline levels of VEGF-C (HR: 0.45; P=0.014) and sVEGFR3 (HR: 0.50; P=0.045). Overall survival was higher in patients with high sVEGFR3 (HR: 0.36; P=0.030) levels. In both responders and non-responders, sVEGFR-1 and PlGF dramatically increased upon exposure to aflibercept and remained overexpressed for the all course of induction therapy. Circulation biomarkers modulations will be analyzed upon the 6 cycles. Tumor tissue biomarker assessment at baseline using IHC and theEGFR pathway mutations status will be added. Conclusions: Exposure to aflibercept is associated with an increase of sVEGFR-1 and PlGF at cycle 1. Elevated baseline expression levels of on-target sVEGFR3 predict favorable outcome in patients treated with aflibercept. Citation Format: Annemilai Tijeras-Raballand, Armand de Gramont, Marielle Chiron, Jean-Baptiste Bachet, Thierry Andre, Dominique Auby, Jerome Desramé, Baba-Hamed Nabil, Lecaille Cedric, Valérie Lebrun, Christophe Louvet, Annette Larsen, Christophe Tournigand, Sylvie Benner, Malika Attia, Aimery de Gramont, Franck Bonnetain, Benoist Chibaudel. Impact of circulating and tissue biomarkers in patients with metastatic colorectal cancer treated with first-line FOLFOX-aflibercept therapy. Results of the GERCOR VELVET Phase II Study [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2734. doi:10.1158/1538-7445.AM2017-2734

Warm and Humidified Versus Cold and Dry CO2 Pneumoperitoneum in Minimally Invasive Colon Resection: A Randomized Controlled Trial.

Peritoneal insufflation with warm-humidified (WH) CO2 gas during minimally invasive surgical procedures is purported to prevent hypothermia and peritoneal desiccation and is associated with decreased postoperative IL-6 levels. This randomized study's purpose was to determine the clinical impact of WH versus cold-dry (CD) CO2 in minimally invasive colon resection (MICR), and to assess perioperative plasma levels of IL-6, TIMP-1, sVEGF-R1, and HSP-70 after MICR. Operative and short-term clinical data plus perioperative blood samples were collected on MICR patients randomized to receive either WH (36.7°C, 95% humidity) or CD (room temperature, 0% humidity) CO2 perioperatively. Peritoneal biopsies were taken at the start and end of surgery. Outcomes tracked included core temperature, postoperative in-hospital pain levels, analgesia requirements, and standard recovery parameters. Preoperative and postoperative days (PODs) 1 and 3 plasma protein levels were determined via ELISA. A total of 101 patients were randomized to WH CO2 (50) or CD CO2 (51). The WH group contained more diabetics ( P = .03). There were no differences in indication, minimally invasive surgical method used, or core temperature. Pain scores were similar; however, the WH patients required less narcotics on PODs 1 to 3 ( P < .05), and less ketorolac on PODs 1 and 2 ( P < .03). No differences in length of stay, complication rates, or time to flatus/diet tolerance were noted. Plasma levels of the 4 proteins were similar postoperatively. Though insignificant, the WH group had less marked histologic changes on "end-of-case" peritoneal biopsies. This study found significantly lower pain medication requirements for PODs 1 to 3 for the WH group; however, because there were no differences in the pains scores between the groups, firm conclusions regarding WH CO2 cannot be made.

Dysregulation of circulating autoantibodies against VEGF-A, VEGFR-1 and PlGF in preeclampsia - A role in placental and vascular health?

Preeclampsia is a state of antiangiogenesis, with high levels of maternal circulating sVEGFR-1 (soluble vascular endothelial growth factor receptor 1, also named sFlt1) and low levels of PlGF (placenta growth factor). Various autoantibodies have been detected in preeclamptic patients. We hypothesize that circulating autoantibodies against VEGF-A (AA-VEGF-A), VEGFR-1 (AA-VEGFR-1) and PlGF (AA-PlGF) are present in preeclamptic women, with different levels from pregnant women with normotensive pregnancies. Secondly, we wanted to analyze if autoantibody levels are associated to sFlt1 or PLGF levels. Retrospective cross sectional study of 88 women with singleton pregnancies who delivered at Oslo University Hospital of whom 46 had preeclampsia and 42 had uncomplicated normotensive pregnancies. Novel immunoassays for IgG-autoantibodies against VEGFA, VEGFR-1 and PlGF were developed and serum samples were assayed. AA-VEGF-A, AA-VEGF-R1 and AA-PlGF were significantly lower in preeclamptic pregnancies (n=42) compared to normotensive pregnancies (n=46) (p<0.05). On unadjusted analysis, only AA-VEGFA and AA-VEGFR-1 were predictors of PE, but none were independent predictors after adjusting for BMI (body mass index) and parity. In the subgroup of normotensive and PE women with overlapping sVEGFR-1/PlGF-ratios, AA-VEGF was a significant predictor of PE with AUC: 0.735. IgG autoantibodies against VEGF-A VEGFR-1 and PlGF can be found in pregnant women. They are dysregulated in preeclampsia. The roles of these autoantibodies are unknown, but this study suggests they play a protective role in pregnancy. The levels of AA against VEGF-A, VEGFR-1 and PlGF might be important factors contributing to anti-angiogenesis regulation.

Aqueous Humor Levels of Soluble Vascular Endothelial Growth Factor Receptor and Inflammatory Factors in Diabetic Macular Edema

Aqueous levels of soluble vascular endothelial growth factor receptor (sVEGFR) and inflammatory factors were measured in 35 patients (37 eyes) with diabetic macular edema (DME) receiving anti-vascular endothelial growth factor (VEGF) therapy. Aqueous levels of growth factors (VEGF, placental growth factor [PlGF], and platelet-derived growth factor AA [PDGF-AA]), sVEGFR-1 and -2, soluble intercellular adhesion molecule 1, monocyte chemotactic protein (MCP)-1, interleukin (IL)-6, -8, -12, and -13, and interferon-inducible 10-kDa protein (IP-10) were significantly higher in the DME group than in the nondiabetic control group. The sVEGFR-2 level was significantly correlated with the neurosensory retinal thickness, as well as with the levels of growth factors (VEGF and PDGF-AA) and inflammatory factors (MCP-1, IL-6, and IL-8). Three growth factors (VEGF, PlGF, and PDGF-AA) were also significantly correlated with each other, as were sVEGFR-1 or -2 and the inflammatory factors (MCP-1, IL-6, IL-8, and IP-10). These findings suggest that sVEGFRs and growth/inflammatory factors have an important role in DME.

Kif4 regulates the expression of VEGFR1 through the PI3K/Akt signaling pathway in RAW264.7 monocytes/macrophages.

Kinesin superfamily protein4(Kif4), a microtubule-based motor protein, has been shown to participate in a number of critical cellular processes, such as cell division, the intracellular transport of membranous vesicles and signal transduction. However, whether KIF4 regulates vascular endothelial growth factor(VEGF) receptor1(VEGFR1) expression remains unknown. Thus, in this study, in order to examine the effects of Kif4 on the expression of VEGFR1 in RAW264.7 monocytes/macrophages, Kif4 was silenced using siRNA. RT-qPCR, western blot analysis and ELISA were used to assess the expression of Kif4 and VEGFR1 up- and downstream signaling molecules, including VEGF-A, VEGFR1, soluble form of VEGFR1(sVEGFR1), phosphorylated(p-)Akt and Akt. The silencing Kif4 inhibited the mRNA expression of VEGF(P<0.01) and p-Akt(P<0.05); however, the level of VEGF-A was increased(P<0.05) compared with the negative control siRNA-transfected group. The silencing of Kif4 decreased the VEGFR1 mRNA(P<0.05), VEGFR1 protein and sVEGFR1 levels in the cell supernatant(P<0.01). Following the application of insulin-like growth factor-1(100ng/ml), the specific agonist of PI3K/Akt in the Kif4siRNA-transfected group,the VEGFR1 mRNA levels(P<0.001), the VEGFR1 protein levels and the sVEGFR1(P<0.01) levels significantly increased; however, the levels of VEGF in the cell supernatant were decreased(P<0.05). Taken together, these findings suggest that Kif4 regulates the expression of VEGFR1 in RAW264.7cells and that the PI3K/Akt pathway is involved in this process.

Angiogenic effects of low-intensity cathodal direct current on ischemic diabetic foot ulcers: A randomized controlled trial.

This study investigated the effect of low-intensity cathodal direct current (CDC) of electrical stimulation (ES) on the release of hypoxic inducible factor-1α (HIF-1α), nitric oxide (NO), vascular endothelial growth factor (VEGF), and soluble VEGF receptor-2 (sVEGFR-2) in the wound fluid of ischemic diabetic foot ulcers (DFUs). This study was a randomized, single-blind, placebo-controlled trial. Thirty type 2 diabetes patients with ischemic foot ulcerations were randomly assigned to receive either low-intensity CDC at sensory threshold (ES group, n=15) or placebo treatment (control group, n=15) for 1h/day, 3days/week, for 4weeks (12 sessions). After debridement during the first and twelfth treatment sessions, wound fluid was collected before and after ES application to determine the levels of HIF-1α, NO, VEGF, and sVEGFR-2. Wound surface area (WSA) was measured at the first, sixth, and twelfth sessions. At the first session, after ES application, wound-fluid levels of HIF-1α were significantly increased (+61.98pg/mL) compared to the control group (-3.85pg/mL, P=0.01). After ES application at the first and twelfth sessions, wound-fluid levels of VEGF were also significantly increased (+36.77 and +39.57pg/mL, respectively) compared to the control group (+4.15 and +0.15pg/mL, P=0.007 and P=0.019, respectively). There was no significant effect on NO and sVEGFR-2 levels between the groups. Low-intensity CDC has positive effects on the release of HIF-1α and VEGF in the wound area of ischemic DFUs. Furthermore, our results suggest that applying ES to ischemic DFUs can be a promising way to promote angiogenesis and to achieve better outcomes in diabetic wound healing.

Inflammatory cytokines and angiogenic factors as potential biomarkers in South African pancreatic ductal adenocarcinoma patients: A preliminary report

Background/ObjectivesSeveral studies have investigated the association of differentially expressed cytokines with pancreatic ductal adenocarcinoma (PDAC), but none in African countries. This study aimed at investigating T-helper (Th) cell and angiogenic markers as diagnostic or prognostic biomarkers for PDAC in Black South Africans. MethodsWe conducted a prospective, case-control study comprising of 34 PDAC patients and 27 control participants with either critical limb ischemia, abdominal aortic aneurysm or other abdominal pathology from causes other than pancreatic disease. Plasma levels of IL-2, IL-4, IL-6, IL-10, TNF, IFN-γ, IL-17A, VEGF, sVEGF-R1, FGF, PIGF, PDGF and P-selectin were measured using commercially available cytometric bead array, ELISA and multi-analyte Luminex kits. ResultsSignificantly higher levels of IFN-γ (p < 0.001), TNF (p < 0.001), IL-2 (p = 0.001), IL-4 (p < 0.01), IL-10 (p < 0.01), IL-17A (p < 0.01), PlGF (p < 0.0001) and basic FGF (p < 0.0001) were found in cases compared to control participants. PDAC patients with irresectable tumours had higher levels of VEGF (p = 0.02) and IL-6 (p = 0.01). A univariate analysis showed significant associations between IFN-γ, TNF, IL-10, -4, -2, basic FGF, PlGF and PDAC. In a multivariate logistic regression model, basic FGF (p = 0.002) and PlGF (p = 0.007) were independent risk factors for PDAC with a combined sensitivity of 71% and specificity of 100%. ConclusionOur preliminary data suggests a potential role for basic FGF and PlGF as diagnostic, and VEGF and IL-6 as prognostic biomarkers of PDAC in Black South African patients.

Serum levels of platelet endothelial cell adhesion molecule-1 (PECAM-1) and soluble vascular endothelial growth factor receptor (sVEGFR)-1 and -2 in HIV associated preeclampsia

ABSTRACTObjective: The angiogenic–antiangiogenic imbalance evident in preeclampsia (PE) may be used as a predictive tool to identify women likely to develop the clinical features in early pregnancy. Method: This retrospective study examined normotensive pregnant (n = 38) and preeclamptic (n = 38) HIV-infected and uninfected women to quantify sVEGFR-1 and -2 and PECAM-1 levels. Results: In contrast to PECAM-1, sVEGFR-1 and -2 differed according to pregnancy type (p = 0.07; p = 0.001; p = 0.002) but not by HIV status (p = 0.68; p = 0.13; p = 0.43). Conclusion: Irrespective of the HIV status, we report an upregulation of sVEGFR-1 with concomitant decline of PECAM-1 and sVEGFR-2 levels in PE compared to normotensive pregnancies.

Diagnostic Values of sVEGFR-1 and Endostatin in Malignant Pleural Effusions in Patients with Lung Cancer.

The diagnosis of malignant pleural effusion (MPE) remains a common clinical challenge because of the sensitivity of conventional cytology for the detection is insufficient. Thus, a sensitive clinical marker for diagnosis is required. The aim of this study was to assess the role of two anti-angiogenic cytokines, soluble vascular endothelial growth factor receptor-1 (sVEGFR-1) and endostatin, in diagnosing MPE. Effusion samples from 44 patients with MPE caused by lung cancer and from 36 patients with benign pleural effusion (BPE) were collected. The concentrations of sVEGFR-1 and endostatin in pleural fluid were determined by an enzyme-linked immunosorbent assay (ELISA). The diagnostic performance was measured by receiver operating characteristic curves (ROCs). The levels of sVEGFR-1 and endostatin in MPE due to lung cancer were significantly higher than those in BPE (p < 0.05). The sensitivity and specificity of endostatin were 52.27% and 86.11%, respectively, while for sVEGFR-1, the sensitivity was 88.64% and the specificity was 58.33%. Interestingly, the combination of sVEGFR1 and endostatin produced better sensitivity and specificity of 72.73% and 83.33%, respectively. In addition, the levels of sVEGFR-1 and endostatin were significantly related to each other (p < 0.05), and the levels of endostatin in bloody effusions were significantly higher than those in non-bloody effusions (p < 0.05). Our study indicated that the levels of sVEGFR-1 and endostatin were significantly elevated in MPE. The combined detection of sVEGFR-1 and endostatin may be useful in the diagnosis of MPE.

Follistatin and Endoglin: Potential Biomarkers of Endothelial Damage and Non-Relapse Mortality after Myeloablative Allogeneic Hematopoietic Cell Transplantation in Blood and Marrow Transplant Clinical Trials Network (BMT CTN) 0402

Introduction:Inflammation and angiogenesis are linked biologic processes influencing the onset and recovery of acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic cell transplantation (HCT). In aGVHD, angiogenic factors (AF) contributing to tissue healing/repair (e.g. epidermal growth factor [EGF]) versus AF associated with tissue damage/inflammation (e.g. follistatin [FS]) exist in a dysregulated balance. Whether these AF are associated with, or predictive of, non-relapse mortality (NRM) in the absence of aGVHD, is not understood. Given that regimen-related tissue injury plays a role in NRM, we hypothesized that elevated levels of inflammation-associated AF early after HCT would be associated with NRM even in patients without aGVHD.Methods:We studied plasma samples from patients enrolled on Blood and Marrow Transplant Clinical Trials Network (BMT CTN) aGVHD prophylaxis study 0402, a randomized trial of tacrolimus/methotrexate vs tacrolimus/sirolimus after matched sibling peripheral blood stem cell transplantation with myeloablative conditioning. Levels of AF (angiopoietin-2 [Ang2], EGF, FS, vascular endothelial growth factor [VEGF]-A and -B, endoglin [sEng], placental growth factor [PlGF], and soluble VEGF receptor [sVEGFR]-1 and -2) were quantified by MILLIPLEX magnetic bead panels.We analyzed factors associated with NRM in univariate analysis and used multivariate analysis to develop composite AF scores to predict NRM. In calculating the composite scores, we extrapolated the individual probabilities of NRM based upon regression models, and sorted the individual probabilities into 3 composite scores based on: 1) estimated probability of NRM <10%, 2) probability of NRM between 10-19%, 3) probability of NRM >20%.Results:221 patients from BMT CTN 0402 had pre-transplant plasma samples and samples at day +28 available for analysis. Univariate analysis showed associations of raw levels of Ang2, FS, sEng, and sVEGFR1 on day +28 with NRM. Of these, only higher levels of log-transformed FS (p=0.01) and of sEng (p=0.04) showed a correlation with NRM; these two AF were selected for the final model. In multivariate analysis, patients with the highest levels of day +28 FS and sEng (score 3, n=32, Figure) had a 4.6-fold higher relative risk (RR) of NRM (95% confidence interval 1.7-12.3, p<0.01, Table). The prognostic value of the composite score was stronger than either individual factor alone (RR log[FS]=2.1, p=0.03, and RR log[sEng]=2.3, p=0.07). Grade III-IV aGVHD and patient age>50 years were also independently associated with 1-year NRM. There was no effect of patient gender, disease status, CMV serostatus, or type of GVHD prophylaxis (tacrolimus/methotrexate versus tacrolimus/sirolimus) on NRM. There was no significant effect of the composite score, grade III-IV aGVHD, or age on the competing risk of relapse. FS and sEng measured before HCT were not associated with 1-year NRM (p=0.91 and p=0.62, respectively). There was only a weak correlation between the day 0 and day 28 FS and sEng (Spearman’s rho 0.36 and 0.38, respectively). Among the 221 cohort patients, there were 25 deaths due to NRM, including 5 deaths occurring prior to day 56. Death was more likely to be from organ toxicity in patients with composite score 3 (5/9, 56%) vs those with a composite score of 1 (1/8, 12%) or 2 (3/8, 37%).Conclusions:A composite score using day 28 plasma levels of FS and sEng predicts the risk of 1-year NRM after HCT, providing further evidence that endothelial dysfunction is a relevant biological problem in HCT. Pre-HCT measurements of FS and sEng were not associated with NRM, suggesting that elevations in these factors may reflect susceptibility to regimen-related toxicity. We have previously shown that elevated levels of FS and sEng at the onset of aGVHD are associated with poor survival. Here, we show that these factors are predictive of NRM independent of aGVHD. Follistatin modulates inflammatory responses following tissue injury via interaction with the transforming growth factor β superfamily and may reflect endothelial cell damage. sEng is upregulated in proliferating endothelial cells, in inflammation and tissue ischemia, and can influence leukocyte adhesion and endothelial transmigration. Measurement of these predictive AF may highlight prospective opportunities to intervene and prevent subsequent complications thereby limiting NRM after allogeneic HCT. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Overweight and obesity versus concentrations of VEGF-A, sVEGFR-1, and sVEGFR-2 in plasma of patients with lower limb chronic ischemia.

Being overweight or obese comprises a significant risk factor for atherosclerosis. Fat tissue also generates factors stimulating angiogenesis, the process by which new blood vessels form. The purpose of this paper is to assess concentrations of the vascular endothelial growth factor A (VEGF-A) and its soluble type-1 and type-2 receptors (sVEGFR-1 and sVEGFR-2) in plasma of patients with peripheral arterial disease (PAD) depending on the level of nutrition according to body mass index (BMI). The study group included patients suffering from symptomatic PAD (n=46) in Fontaine classes IIa-IV without any history of neoplastic disease and who have a normal BMI (n=15), are overweight (n=21) or are obese (n=10). The control group (n=30) consisted of healthy non-smoking volunteers who were neither overweight nor obese. Venous blood plasma samples were collected from both groups at rest in the morning to determine plasma concentrations of VEGF-A, sVEGFR-1, and sVEGFR-2 using the enzyme-linked immunosorbent assay (ELISA) method. The group of patients with PAD co-existent with being overweight or obese tended to have higher mean concentration levels of VEGF-A and sVEGFR-2 when compared with patients suffering from PAD with normal BMI. A statistically significant positive correlation was obtained between BMI and average plasma concentrations of sVEGFR-2 (R=0.37, P=0.0103). However, no significant correlation was noticed between BMI and VEGF-A or sVEGFR-1 concentrations. A positive correlation determined between the level of antiangiogenic factor and BMI value may be indicative of the linearly growing prevalence of some antiangiogenic factors in patients with metabolic disorders, which may be one of numerous factors contributing to incomplete efficiency of collateral circulation development in patients with PAD.

Soluble vascular endothelial growth factor receptor-1 is reduced in patients with resistant hypertension after renal denervation

Renal denervation (RDN) has been shown in several studies to reduce blood pressure (BP) in patients with resistant hypertension (RH). Data on potential biomarkers associated with BP changes remain scarce. We evaluated whether soluble vascular endothelial growth factor receptor (sVEGFR-1) is affected by the procedure. A total of 57 patients with RH participated in this study. BP and heart rate were recorded at baseline and at 3 months follow-up, at which time blood samples were collected to determine the levels of sVEGFR-1, VEGF-A, VEGF-C, nitric oxide (NO), soluble vascular adhesion molecule 1 and soluble intracellular adhesion molecule 1. None of the biomarkers had a predictive value that could identify responders vs non-responders to RDN. However, sVEGFR-1 concentration was dramatically reduced after RDN (5913±385 vs 280±57 pg ml-1, P<0.001). At the same time VEGF-A levels were significantly increased (10.0±3.0 vs 55.5±7.9 pg ml-1, P<0.001), without significant changes in VEGF-C. NO levels were significantly increased after RDN in the whole group (82.6±6.2 vs 106.9±7.8 μM, P=0.021). Interestingly, the elevation in NO levels at 3 months was only seen in patients who demonstrated a reduction in systolic BP of ⩾10 mm Hg (78.9±8.3 vs 111.6±11.7 μM, P=0.018). We report a significant reduction in sVEGFR-1 levels after RDN procedure, which was accompanied by a significant increase in VEGF-A concentration as well as NO. Changes in plasma cytokines were not quantitatively linked to magnitude of BP reduction. An RDN-induced reduction in sVEGFR-1 plasma levels and increase in VEGF-A would raise the VEGF-A/sVEGFR-1 ratio, thereby increasing VEGF-A bioavailability to act on its full-length receptor and may contribute to the BP-lowering effect potentially via NO-mediated pathways.

POSSIBLE MOLECULAR BASIS OF BEVACIZUMAB THERAPY FOR MACULAR EDEMA IN BRANCH RETINAL VEIN OCCLUSION.

To investigate changes in the aqueous humor levels of 11 factors/cytokines (including vascular endothelial growth factor [VEGF] receptors, growth factors, and inflammatory factors) after intravitreal injection of bevacizumab (IVB), as well as the relationship between changes of these factors and improvement of macular edema in patients with branch retinal vein occlusion and macular edema. In 29 patients with branch retinal vein occlusion who received IVB twice for macular edema at monthly intervals, aqueous humor samples were obtained during IVB. Levels of VEGF, soluble VEGF receptor (sVEGFR)-1, sVEGFR-2, placental growth factor, soluble intercellular adhesion molecule-1, monocyte chemotactic protein 1, platelet-derived growth factor-AA, and interleukin (IL)-6, IL-8, IL-12 (p70), and IL-13 were measured by the suspension array method. Foveal thickness was examined by optical coherence tomography before and 1 month after IVB, and the improvement of macular edema was evaluated by calculating the percent change of foveal thickness. Aqueous humor levels of sVEGFR-1, VEGF, monocyte chemotactic protein 1, and IL-6 showed a significant decrease at 1 month after IVB compared with baseline (P = 0.013, P < 0.001, P = 0.047, and P = 0.019, respectively). There was a significant negative correlation between the change of sVEGFR-1 or platelet-derived growth factor-AA after IVB and the improvement of macular edema (P = 0.004 and P = 0.036, respectively). These findings suggest that improvement of macular edema by IVB is related to inhibition of sVEGFR-1 and platelet-derived growth factor-AA, but not VEGF, in patients with branch retinal vein occlusion. Soluble VEGF receptor-1 and platelet-derived growth factor-AA might be useful indicators of the response of macular edema.