sVEGFR-1 Levels Research Articles

Circulating endothelial cell number and markers of endothelial dysfunction in previously preeclamptic women

Patients with preeclampsia (PE) have endothelial dysfunction and an increased future risk of cardiovascular (CV) mortality. The number of circulating endothelial cells (CECs) is markedly increased in conditions associated with a high degree of endothelial cell activation/injury including PE. We hypothesized that the number of CECs continues to be increased in women with a history of PE, reflecting ongoing endothelial cell activation/injury. CECs, flow-mediated vasodilation, levels of adhesion molecules and soluble vascular endothelial growth factor receptor-1 (sVEGFR1), and urine albumin/creatinine ratio were determined in 21 healthy women with ongoing normal pregnancy, 24 healthy currently nonpregnant women with a history of normal pregnancy, a total of 17 women with currently active mild (n = 11) or severe (n = 6) PE without hemolysis, elevated liver enzymes, and low platelet count (HELLP) syndrome, and 16 currently nonpregnant women with a history of mild (n = 10) or severe (n = 6) PE. Blood samples from women with active preeclampsia had higher CECs (9.9 ± 7.9 cells/mL) than healthy pregnant women (3.0 ± 4.1 cells/mL; P < .001), healthy nonpregnant women with a history of normal pregnancy (3.4 ± 4.0 cells/mL; P < .001), or women with a history of preeclampsia (2.4 ± 2.0 cells/mL; P < .001). The number of CECs were similar between women with a history of preeclampsia and healthy nonpregnant women with a history of normal pregnancy. Patients with active preeclampsia had significantly higher soluble vascular cell adhesion molecule-1, soluble E-selectin, sVEGFR1, and urinary albumin/creatinine ratio than healthy pregnant women. However, soluble vascular cell adhesion molecule-1, soluble E-selectin, urinary albumin/creatinine ratio were similar in women with a history of preeclampsia and healthy nonpregnant women with a history of normal pregnancy. However, women with a history of preeclampsia had higher sVEGFR1 levels than women with a history of normal pregnancy (P < .05). Markers of endothelial activation, dysfunction, and damage were increased in patients with PE. After the delivery, this activation status is similar to the age-matched nonpregnant women with a history of normal pregnancy. However, sVEGFR-1 levels remain higher in women with a history of preeclampsia compared with women without a history of preeclampsia.

Circulating cytokines and monocyte subpopulations as biomarkers of outcome and biological activity in sunitinib-treated patients with advanced neuroendocrine tumours

Background:Sunitinib is approved worldwide for treatment of advanced pancreatic neuroendocrine tumours (pNET), but no validated markers exist to predict response. This analysis explored biomarkers associated with sunitinib activity and clinical benefit in patients with pNET and carcinoid tumours in a phase II study.Methods:Plasma was assessed for vascular endothelial growth factor (VEGF)-A, soluble VEGF receptor (sVEGFR)-2, sVEGFR-3, interleukin (IL)-8 (n=105), and stromal cell-derived factor (SDF)-1α (n=28). Pre-treatment levels were compared between tumour types and correlated with response, progression-free (PFS), and overall survival (OS). Changes in circulating myelomonocytic and endothelial cells were also analysed.Results:Stromal cell-derived factor-1α and sVEGFR-2 levels were higher in pNET than in carcinoid (P=0.003 and 0.041, respectively). High (above-median) baseline SDF-1α was associated with worse PFS, OS, and response in pNET, and high sVEGFR-2 with longer OS (P⩽0.05). For carcinoid, high IL-8, sVEGFR-3, and SDF-1α were associated with shorter PFS and OS, and high IL-8 and SDF-1α with worse response (P⩽0.05). Among circulating cell types, monocytes showed the largest on-treatment decrease, particularly CD14+ monocytes co-expressing VEGFR-1 or CXCR4.Conclusions:Interleukin-8, sVEGFR-3, and SDF-1α were identified as predictors of sunitinib clinical outcome. Putative pro-tumorigenic CXCR4+ and VEGFR-1+ monocytes represent novel candidate markers and biologically relevant targets explaining the activity of sunitinib.

Circulating cytokines and angiogenic factors (CAFs) in patients with advanced non-clear cell renal cell carcinoma treated with sunitinib: Results from a phase II clinical trial.

483 Background: Development of hypertension (HTN) during VEGF-directed therapies is associated with improved overall survival (OS) in patients (pts) with clear-cell renal cell carcinoma. There is paucity of data about the molecular determinants of response and survival outcomes in non-clear cell renal cell carcinoma (nccRCC) pts treated with targeted therapies. We investigated CAFs and correlated them and HTN with outcomes of nccRCC pts treated with sunitinib in a phase II trial. Methods: Using a bead-based multiplex assay (Luminex), levels of 30 CAFs (including mediators of inflammation, angiogenesis, immunity, and metastasis) were measured prior to start of sunitinib therapy (baseline) and at 4 weeks after completion of cycle 1. Sunitinib benefit was defined as having partial response (PR) or stable disease (SD) by RECIST lasting ≥4 months. Cox proportional hazards regression models were used to assess the association between CAFs, HTN, and survival outcomes. The Kaplan-Meier method was used to estimate progression-free survival (PFS) and OS. Results: Fifty-seven pts [papillary (27), chromophobe (5), unclassified (8), collecting duct/medullary carcinoma (6), sarcomatoid (7), others (4)] were evaluable. Median follow up was 59.1 months (range, 30.6-82.6); 43 pts (75%) have died and 14 (25%) were alive at last follow up. Sunitinib benefit was seen in 21 pts (8 papillary, 4 chromophobe, 3 unclassified, 3 others, 2 collecting duct/medullary carcinoma, 1 sarcomatoid); 2 pts were not evaluable for response. Median PFS was 2.9 months (95% CI, 1.4-5.5) and median OS 16.8 months (95% CI, 10.7-27.4). 29 pts developed HTN during the first cycle, but there was no significant association between HTN and OS, PFS or sunitinib benefit. Elevated baseline levels of GRO, TGFα, sTNF-RI, sVEGF-R2 and IL-8 were correlated with higher risk of death. Conclusions: In this phase II trial in pts with advanced nccRCC, we found no significant association between HTN and PFS, OS, or sunitinib benefit. Candidate CAFs were identified that could be incorporated into prognostic and predictive models for validation in future studies in nccRCC.

Role of soluble vascular endothelial growth factor receptor signaling and other factors or cytokines in central retinal vein occlusion with macular edema.

The association between aqueous humor levels of various growth/inflammatory factors or cytokines and the severity of macular edema was investigated in patients with central retinal vein occlusion (CRVO). Aqueous humor levels of 11 factors or cytokines were measured in CRVO patients with macular edema (n = 38) and in cataract patients (n = 15) as controls. Aqueous humor samples were obtained from the patients during surgery and the levels of VEGF, its receptors, and other factors or cytokines were measured by the suspension array method. The severity of macular edema was determined by measuring the central macular thickness, neurosensory retinal thickness, and subfoveal serous retinal thickness with optical coherence tomography. Aqueous humor levels of growth factors, soluble VEGF receptor 1 (sVEGFR-1), sVEGFR-2, and inflammatory factors or cytokines were significantly higher in the CRVO group than in the control group. The aqueous level of sVEGFR-1 was significantly correlated with the neurosensory retinal thickness, as well as with the levels of growth factors (VEGF, placental growth factor, and platelet-derived growth factor-AA) and inflammatory factors/cytokines (monocyte chemotactic protein-1, interleukin-6, and interleukin-8). Aqueous humor levels of the three growth factors also were significantly correlated with each other, as were levels of the two sVEGFRs. These findings suggest that growth/inflammatory factors and cytokines have an important role in macular edema associated with CRVO. Better understanding of the mechanisms involved may lead to development of new treatments, such as anti-VEGFR-1 therapy.

Phase I trial of sorafenib following liver transplantation in patients with high-risk hepatocellular carcinoma.

401 Background: Liver transplantation (LT) offers excellent long-term survival for hepatocellular carcinoma (HCC) patients who are within established criteria. For those outside such criteria, or with high risk pathologic features in the explant, HCC recurrence rates after LT are high. No treatment has been shown to decrease risk of recurrence post-LT. We conducted a multicenter phase I trial of sorafenib in LT patients with high-risk HCC. Methods: Subjects had pathologically proven high-risk HCC defined as outside Milan (pre- or post-transplant), poorly differentiated tumors, or tumors with vascular invasion. We used a standard 3+3 phase I design, beginning drug between 4 and 16 weeks after LT, with planned duration of treatment of 24 weeks. Cohort dosages were: 1) 200 mg per day, 2) 200 mg twice a day, 3) 200 mg/400 mg per day 4) 400 mg BID. Correlative studies included circulating endothelial cells (CECs) and plasma biomarkers collected prior to treatment, at 1 month, and at recurrence in a subset of subjects, and tumor expression of p-Erk, p-Akt, and c-Met in tissue microarrays. Results: We enrolled 14 patients. Median age was 63 years, and 93% were men. 71% had underlying HCV and 21% had HBV. Maximum tolerated dose (MTD) was 200 mg BID; only 43% of patients received &gt;80% of planned dose. Grade 3-4 toxicities seen in &gt;10% of subjects included: leukopenia (29%), LFT abnormalities (21%), hypertension (14%), hand-foot syndrome (14%) and diarrhea (14%). Over a median follow-up of 953 days, 1 patient died and 4 recurred, with a median recurrence-free survival of 716 days for the 4 patients who recurred. Mean CEC number at baseline was 21 cells/4 ml for those who recurred, and 80 cells/4 ml for those who did not (p=0.10). Mean sVEGFR2 levels decreased after 1 month on sorafenib (p=0.09), but did not correlate with recurrence. There was a trend for tumor c-Met expression with increased risk of recurrence. Conclusions: Post-transplant sorafenib is feasible and tolerable at 200 mg PO BID. Recurrence-free survival appears longer than expected but needs further validation in a larger study. Clinical trial information: NCT00997022.

Analysis of serum protein biomarkers, circulating tumor DNA, and dovitinib activity in patients with tyrosine kinase inhibitor-refractory gastrointestinal stromal tumors

An exploratory translational analysis was conducted as part of a phase II study of dovitinib to assess the relevance of soluble serum proteins and circulating tumor (ct) DNA (ctDNA) as biomarkers in patients with tyrosine kinase inhibitor (TKI)-refractory gastrointestinal stromal tumors (GISTs). Predose serum samples were collected from 30 patients on day 1 of cycle 1 and cycle 2. Serum levels of angiogenesis-related proteins were assessed by enzyme-linked immunosorbent assay, and Beads, emulsions, amplification, and magnetics (BEAMing) assays were carried out to detect mutations in serum ctDNA. Dovitinib increased vascular endothelial growth factor (VEGF)165 (1.26-fold, P = 0.006), VEGF-A (1.27-fold, P = 0.004), placental growth factor (6.0-fold, P = 0.002), fibroblast growth factor 23 (1.45-fold, P = 0.02), and interleukin 8 (1.75-fold, P = 0.04) levels, and decreased soluble vascular endothelial growth factor receptor (sVEGFR)-2 levels (0.8-fold, P = 0.001). The changes in sVEGFR-2 were significantly associated with metabolic response determined by positron emission tomography (P = 0.02) and progression-free survival (PFS; P = 0.02). Secondary kinase mutations were identified in the ctDNA of 11 patients (41%), and these patients all had mutations involving KIT exon 17. Patients with secondary KIT mutations had significantly worse overall survival {median, 5.5 months [95% confidence interval (CI) 3.8-7.2 months]} than those with no detectable secondary mutations [9.8 months (95% CI 9.6-10.0 months); hazard ratio = 2.7 (95% CI 1.0-7.3); P = 0.047]. Changes in sVEGFR-2 levels were associated with dovitinib-mediated antitumor activity. Genotyping of serum ctDNA with BEAMing is useful for the identification of resistant mutations potentially associated with poor prognosis in patients with GISTs.

Role of Inflammation in Diabetic Macular Edema

Vitreous fluid levels of vascular endothelial growth factor (VEGF), soluble VEGF receptor (sVEGFR)-2, soluble intercellular adhesion molecule (sICAM)-1, monocyte chemotactic protein (MCP)-1 and pentraxin 3 (PTX3) were measured by enzyme-linked immunosorbent assay in 36 patients with diabetic macular edema (DME) and 15 patients with macular hole (MH). Aqueous flare values were measured with a laser flare meter, and macular edema was examined by optical coherence tomography. Vitreous fluid levels of VEGF, sVEGFR-2, sICAM-1, MCP-1 and PTX3 were significantly higher in the patients with DME than in those with MH. There was a significant correlation between the vitreous fluid level of sVEGFR-2 and the levels of sICAM-1, MCP-1 and PTX3. The aqueous flare value was significantly correlated with the vitreous fluid levels of sVEGFR-2, sICAM-1, MCP-1 and PTX3. These findings suggest that inflammatory factors may induce an increase in vascular permeability and disrupt the blood-aqueous barrier in DME patients.

Age-Related Changes in Soluble Vascular Endothelial Growth Factor Receptor 1 (sVEGFR1) and Receptor 2 (sVEGFR2) in Healthy Japanese Subjects.

We measured serum soluble vascular endothelial growth factor receptor 1 (sVEGFR1) and receptor 2 (sVEGFR2) levels in healthy Japanese individuals in order to establish a reference value using a specific ELISA. Significant differences were observed in serum sVEGFR1 and sVEGFR2 levels between children and adults. To demonstrate the usefulness of the reference value for children, we measured serum sVEGFR1 and sVEGFR2 levels in children with diarrhea positive (D+) hemolytic uremic syndrome (HUS) as a preliminary study. Serum sVEGFR2 levels in children with HUS were markedly higher than those in healthy children from the onset of D+HUS. The reference value for healthy children in the present study will allow normal and pathological conditions to be discriminated from each other in future study.

Effect of laser photocoagulation on plasma levels of VEGF-A, VEGFR-2, and Tie2 in infants with retinopathy of prematurity

To report the baseline plasma levels of vascular endothelial growth factor-A (VEGF-A), soluble vascular endothelial growth factor receptor-2 (sVEGFR-2) and soluble Tie2 in infants with treatment-requiring retinopathy of prematurity (ROP) and to investigate the effect of laser treatment on the plasma levels. Blood samples were collected from infants with prethreshold type 1 ROP before, 1 day after, and 1 week after confluent laser photocoagulation. Enzyme-linked immunosorbent assay procedure was used to determine plasma VEGF-A, sVEGFR-2, and sTie2 levels. A total of 48 eyes of 30 infants were included. The mean postconceptional age at which laser photocoagulation was performed was 37.5 ± 2.9 weeks. The mean number of laser spots applied to each eye was 1,480 ± 420. Regression of active retinopathy occurred within 1 week of treatment in all cases. Baseline plasma VEGF-A, sVEGFR-2, and sTie2 levels did not differ between unilateral and bilateral and also zone I and zone II disease. A significantly progressive decrease was observed in plasma VEGF-A, sVEGFR-2, and sTie2 levels at 1 day and 1 week after laser photocoagulation. Plasma levels of VEGF-A, sVEGFR-2, and sTie2 decreased significantly in our patients after laser treatment; however, because of the variability in the measurements, further studies evaluating the clinical value of these angiogenic factors in patients with treatment-requiring ROP are necessary.

Interleukin-4 and granulocyte-macrophage colony-stimulating factor mediates the upregulation of soluble vascular endothelial growth factor receptor-1 in RAW264.7 cells-a process in which p38 mitogen-activated protein kinase signaling has an important role.

Soluble vascular endothelial growth factor receptor-1 (sVEGFR1) antagonizes angiogenesis by inhibiting the biological function of vascular endothelial growth factor (VEGF). Immature dendritic cells (imDCs) express high levels of sVEGFR1 during development and are antiangiogenic. This study aimed to investigate the changes in VEGFR1, sVEGFR1, and VEGF levels during the development of imDCs and explore the underlying signaling mechanisms. To model the differentiation of imDCs from monocytes, RAW264.7 cells, a murine monocyte/macrophage cell line, were stimulated by interleukin-4 (IL-4; 10ng/mL, 20ng/mL, and 40ng/mL) and/or by granulocyte-macrophage colony-stimulating factor (GM-CSF; 10ng/mL, 20ng/mL, and 50ng/mL) and/or pretreated by the p38 inhibitor SB203580. The levels of VEGFR1, sVEGFR1, and VEGF were detected by reverse transcription polymerase chain reaction (RT-PCR), Western blot, and enzyme-linked immunosorbent assay (ELISA). IL-4 increased the VEGFR1 mRNA and sVEGFR1 levels in RAW264.7 (p<0.05). This increase was inhibited by SB203580. Granulocyte-macrophage colony-stimulating factor increased the sVEGFR1 levels, but it had no significant effect on VEGFR1 mRNA levels. SB203580 decreased the expression of VEGFR1 mRNA induced by GM-CSF, whereas sVEGFR1 was unaffected. IL-4 had a greater effect on sVEGFR1 levels, compared to GM-CSF. IL-4 and GM-CSF increased sVEGFR1 levels, but did not significantly effect VEGF expression, and led to the antiangiogenesis properties of monocytes. p38 Mitogen-activated protein kinase signaling has an important role in the process.

Maternal/newborn VEGF-C936T interaction and its influence on the risk, severity and prognosis of preeclampsia, as well as on the maternal angiogenic profile

Objective: To analyze the influence of maternal/newborn vascular endothelial growth factor (VEGF)-CT936 interaction as a modulating factor in preeclampsia as well as its influence on the maternal angiogenic balance.Methods: Seventy pairs of preeclamptic women/newborns and 94 pairs of normal pregnant mothers/newborns were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Serum VEGF and soluble VEGF receptor-1 (sVEGFR-1) levels were measured using ELISA.Results: The risk to develop mild (odds ratio; OR: 3.79, p = 0.008) and severe (OR: 2.94, p = 0.037) preeclampsia being increased in association with the CT936-VEGF genotype and increased in severe preeclampsia to 6.07 (p = 0.03) if the women were carriers of the homozygous TT936-VEGF genotype. The presence of the VEGF-T936 allele in both the mother and the newborn significantly increases the risk of pregnancy-induced hypertension (PIH), mild and severe preeclampsia. If both the mothers and newborns were carriers of the VEGF-T936 allele, significantly lower VEGF and higher sVEGFR-1 levels were observed for all types of preeclampsia. Pregnant women with PIH and severe preeclampsia delivered at a significantly earlier gestational age neonates with a significantly lower birth weight if both the preeclamptic mothers and their newborns were carriers of the VEGF-T936 allele.Conclusions: Our study suggests the role of maternal/fetal VEGF-CT936 polymorphism as a modulating factor in preeclampsia, which affects the angiogenic balance in preeclamptic mothers, as well as their pregnancy outcome.

Vascular Endothelial Growth Factor and Its Soluble Receptors-1 and -2 in Iris Neovascularization and Neovascular Glaucoma

Concentrations of vascular endothelial growth factor (VEGF), soluble VEGF receptor-1 (sVEGFR-1) and soluble VEGF receptor-2 (sVEGFR-2) were measured by enzyme-linked immunosorbent assay in 27 proliferative diabetic retinopathy patients who had iris neovascularization (INV) with or without neovascular glaucoma (NVG). The 27 eyes were divided into two groups, consisting of INV without elevation of the intraocular pressure (IOP; INV group) and INV with elevated IOP (NVG group). Ten patients with an idiopathic macular hole were used as the controls. The vitreous levels of VEGF, sVEGFR-1 and sVEGFR-2 showed a significant increase across the three groups. In the INV and NVG groups, the vitreous level of VEGF showed a significant correlation with the levels of sVEGFR-1 and sVEGFR-2. There was also a significant correlation between the vitreous levels of sVEGFR-1 and sVEGFR-2. These results suggest that the vitreous levels of sVEGFR-1 and sVEGFR-2 are dependent on VEGF in patients who have INV with or without NVG.

Role of soluble vascular endothelial growth factor receptors-1 and -2, their ligands, and other factors in branch retinal vein occlusion with macular edema.

To evaluate the association between multiple factors in aqueous humor and the severity of macular edema in patients with branch retinal vein occlusion (BRVO). We measured the aqueous humor levels of 11 factors (including vascular endothelial growth factor receptors, growth factors, and inflammatory factors) in BRVO patients with macular edema and in cataract patients as controls. Aqueous humor samples were obtained from 40 patients (31 patients with BRVO and 9 with cataract). Then the levels of vascular endothelial growth factor (VEGF), soluble VEGF receptor (sVEGFR)-1, sVEGFR-2, placental growth factor (PlGF), soluble intercellular adhesion molecule (sICAM)-1, monocyte chemotactic protein (MCP)-1, platelet-derived growth factor (PDGF)-AA, interleukin (IL)-6, IL-8, IL-12(p70), and IL-13 were measured by the suspension array method. Macular edema was examined by optical coherence tomography, and its severity was determined from the central macular thickness (CMT), neurosensory retinal thickness (TNeuro), and subfoveal serous retinal thickness (SRT). Aqueous humor levels of growth factors, sVEGFR-1, sVEGFR-2, and inflammatory factors were significantly higher in eyes with BRVO than in control eyes. Aqueous levels of sVEGFR-1 and -2 were significantly correlated with the SRT, as well as with the levels of growth factors (PIGF and PDGF-AA) and various inflammatory factors (sICAM-1, MCP-1, IL-6, and IL-8). Levels of the growth factors (VEGF, PlGF, and PDGF-AA) were also significantly correlated with each other. These findings suggest the importance of the cytokine network in BRVO patients, and may contribute to understanding the mechanism of macular edema associated with BRVO and to development of new treatments.

Role of inflammation in previously untreated macular edema with branch retinal vein occlusion.

BackgroundThe association of inflammatory factors and the aqueous flare value with macular edema in branch retinal vein occlusion (BRVO) patients remains unclear. The relationship between the aqueous flare value and the vitreous fluid levels of vascular endothelial growth factor (VEGF), interleukin (IL)-6, monocyte chemotactic protein (MCP)-1, soluble intercellular adhesion molecule 1 (sICAM-1), and soluble VEGF receptor-2 (sVEGFR-2) was evaluated to investigate the role of inflammation in BRVO associated with macular edema. Aqueous flare values and the vitreous levels of VEGF, IL-6, MCP-1, sICAM-1, and sVEGFR-2 were compared between previously untreated patients with BRVO and patients with macular hole (MH).MethodsVitreous samples were obtained from 45 patients during vitreoretinal surgery (28 patients with BRVO and 17 with MH), and the levels of VEGF, IL-6, MCP-1, sICAM-1, and sVEGFR-2 were measured by enzyme-linked immunosorbent assay. Retinal ischemia was evaluated by measuring the area of capillary non-perfusion using fluorescein angiography and the Scion Image program. Aqueous flare values were measured with a laser flare meter and macular edema was examined by optical coherence tomography.ResultsThe median aqueous flare value was significantly higher in the BRVO group (12.1 photon counts/ms) than in the MH group (4.5 photon counts/ms, P < 0.001). There were significant correlations between the aqueous flare value and the vitreous levels of VEGF, IL-6, MCP-1, and sICAM-1 in the BRVO group (ρ = 0.54, P = 0.005; ρ = 0.56, P = 0.004; ρ = 0.52, P = 0.006; and ρ = 0.47, P = 0.015, respectively). The aqueous flare value was also significantly correlated with the foveal thickness in the BRVO group (ρ = 0.40, P = 0.037).ConclusionsInflammation may induce an increase of vascular permeability and disrupt the blood-aqueous barrier via release of inflammatory factors (VEGF, IL-6, MCP-1, and sICAM-1) in BRVO patients with macular edema.

Expression of circulating vascular endothelial growth factor-antagonizing cytokines and vascular stabilizing factors prior to and following bypass surgery in patients with moyamoya disease.

The aim of the present study was to investigate the levels of vascular endothelial growth factor (VEGF)-antagonizing cytokines and VEGF-influenced vascular stabilizing cytokines in patients with moyamoya disease (MMD) and the association with postoperative collateral vessel formation. The study population included 53 MMD patients that had undergone indirect bypass surgery and 50 healthy controls. Serum levels of VEGF, thrombospondin-1 (TSP-1), TSP-2, soluble VEGF receptor-1 (sVEGFR-1), sVEGFR-2, endostatin, angiopoietin-1 (Ang-1) and Ang-2 were measured at the baseline (preoperative) and at day seven following surgery. Postoperative collateralization assessment was conducted upon the six-month follow-up cerebral angiography. Cytokine levels were compared between patients with good or poor collateral formation. Compared with the healthy controls, MMD patients exhibited lower baseline levels of sVEGFR-1 (P<0.0001) and sVEGFR-2 (P<0.0001), but higher VEGF expression (P<0.0001). Ang-1 and Ang-2 levels did not exhibit any difference between the two groups. On day seven following surgery, MMD patients exhibited an almost unchanged sVEGFR-1 and sVEGFR-2 expression level, but upregulated expression of VEGF (P<0.0001), Ang-1 (P<0.0001) and TSP-2 (P<0.0001). The six-month follow-up angiographies revealed that 21 patients (45.65%) that had undergone the same surgical procedure achieved good collateralization. Patients with good collateral formation appeared to have lower sVEGFR-1 and sVEGFR-2 levels prior to (P=0.029 and P=0.045, respectively) and at day seven (P=0.044 and P=0.047, respectively) following bypass surgery when compared with the patients with worse collateralization. Therefore, sVEGFR-1 and sVEGFR-2 may play a role in the pathogenesis of MMD. Lower levels of sVEGFR-1 and sVEGFR-2 indicated better postoperative collateralization in the six months following indirect bypass surgery. However, Ang-1 and Ang-2 may not be specifically involved in the course of MMD.

The effect of Silymarin on VEGF, VEGFR-1 and IL-1α levels in placental cultures of severe preeclamptic women.

The purpose of this study was to evaluate the effects of Silymarin on vascular endothelial growth factor (VEGF), soluble VEGF Receptor-1 (sVEGFR-1) and Interleukin-1 alpha (IL-1α) levels in placental tissue samples of severely preeclamptic women. We conducted an in vitro study in Başkent University Faculty of Medicine, Ankara, Turkey between September 2008 and May 2009. A total of 16 placental tissue samples (8 from severe preeclamptic, and 8 from controls) were analysed. Placental samples were incubated, and VEGF, sVEGFR-1, and IL1-α were measured in culture media using an ELISA kit. The effect of Silymarin on these levels was investigated. Descriptive statistics were initially performed, followed by Mann Whitney U-test and Kruskal-Wallis test to compare means between groups. P values less than 0.05 were considered statistically significant. Eight patients were included in the severe preeclampsia (SP) group, whereas the remaining 8 patients were included in the control group. There were no significant correlations between gestational age and placental VEGF, sVEGFR-1 and IL-1α after 48 or 72 hours of incubation. Basal VEGF levels were lower in the SP group; however, it did not reach statistical significance. sVEGFR-1 and IL-1α levels were also similar between the SP and control groups (p>0.05). After 48 and 72 hours of incubation, sVEGFR-1 levels in Silymarin-added SP and control placental cultures were lower than in the samples without Silymarin addition; however, this difference also did not reach significance. Although we could not demonstrate a significant effect on placental cytokines, considering the role of vasospasm, inflammation, angiogenesis, endothelial cell activation, and oxidative stress in preeclampsia, the potential benefits of Silymarin should be evaluated in future trials with a larger sample size.

Multicenter phase I trial of sorafenib (S) in high-risk hepatocellular carcinoma (HCC) patients after liver transplantation (LT).

285 Background: LT offers excellent long-term survival for HCC patients within the Milan criteria. For those outside Milan, recurrence rates are higher. No known treatments have been shown to decrease risk of recurrence post-transplant. Methods: Subjects had pathologically proven HCC that was outside Milan (pre- or post-transplant), was poorly differentiated, or had vascular invasion. We used a standard 3+3 design, beginning drug between 4 and 16 weeks after LT, with planned duration of treatment of 24 weeks. Cohort dosages were: 1) 200 mg q day, 2) 200 mg BID, 3) 200 mg/400 mg q day. Correlative studies included circulating endothelial cells (CECs-Veridex) and plasma biomarkers (VEGF, sVEGFR1, sVEGFR2, IL-6 and HGF) collected prior to treatment, at 1 month, and at recurrence. Results: We enrolled 14 patients; 93% were men. Median age was 63; 71% had underlying HCV, 21% HBV, and 21% ETOH. 57% were outside Milan on preoperative imaging. On explant, 29% had poorly-differentiated tumors, and 36% had vascular invasion. MTD was 200 mg BID; only 43% of patients received &gt;80% of planned dose. DLTs included liver function abnormalities (1), transplant rejection leading to death (1), and grade 3 hypertension (1). Grade 3-4 toxicities seen in &gt;10% of subjects included: leukopenia (29%), LFT abnormalities (21%), hypertension (14%), hand-foot syndrome (14%) and diarrhea (14%). Over a median follow-up of 836 days, 1 patient died and 4 recurred (median time to recurrence=716 days). Two recurrences were in subjects with limited exposure to S. Median CEC number at baseline was 16 cells/4 ml for those who recurred, and 42 cells/4 ml for those who did not (p=0.10). Median sVEGFR2 levels decreased after 1 month on S (p=0.09), but did not predict recurrence. Conclusions: To our knowledge, this is the first multicenter phase I trial of S completed in the post-transplant setting for high-risk HCC. MTD of S was 200 mg PO BID. Post-transplant S was feasible. Exploratory biomarker data suggest that low CECs at baseline were potentially associated with a higher recurrence rate. Time to recurrence was promising compared with historic controls, but needs to be further explored and validated in a larger study. Clinical trial information: NCT00997022.

A comparison of adipose and bone marrow-derived mesenchymal stromal cell secreted factors in the treatment of systemic inflammation

BackgroundBone marrow-derived mesenchymal stromal cells (BMSCs) are a cell population of intense exploration for therapeutic use in inflammatory diseases. Secreted factors released by BMSCs are responsible for the resolution of inflammation in several pre-clinical models. New studies have uncovered that adipose tissue also serves as a reservoir of multipotent, non-hematopoietic stem cells, termed adipose-derived stromal/stem cells (ASCs), with many common characteristics to BMSCs. We hypothesized that ASC and BMSC secreted factors would lead to a comparable benefit in the context of generalized inflammation.FindingsProteomic profiling of conditioned media revealed that BMSCs express significantly higher levels of sVEGFR1 and sTNFR1, two soluble cytokine receptors with known therapeutic activity in sepsis. In a prophylactic study of endotoxin-induced inflammation in mice, we observed that BMSC secreted factors provided a greater survival benefit and tissue protection of endotoxemic mice compared to ASCs. Neutralization of sVEGFR1 and sTNFR1 did not significantly affect the survival benefit experienced by mice treated with BMSC secreted factors.ConclusionsOur findings suggest that BMSCs may be more effective as a cell therapeutic for use in endotoxic shock and that ASCs may be positioned for continued exploration in immunomodulatory diseases. Soluble cytokine receptors can distinguish stromal cells from different tissue origins, though they may not be the sole contributors to the therapeutic benefit of BMSCs. Furthermore, other secreted factors not discussed in this study may also differentiate these stromal cell populations from one another.

Increased Serum Level of Soluble Vascular Endothelial Growth Factor Receptor-1 Is Associated With Poor Coronary Collateralization in Patients With Stable Coronary Artery Disease

The present study investigated whether serum levels of soluble vascular endothelial growth factor receptor (sVEGFR)-1, -2 and -3 are related to poor coronary collateralization in patients with stable coronary artery disease (CAD). Serum levels of sVEGFR-1, -2, -3, VEGF, and placental growth factor (PLGF) were determined in 403 consecutive patients with angiographic total or subtotal occlusion of at least 1 major coronary artery. The degree of collateralization was graded according to the Rentrop scoring system. Low (Rentrop score of 0 or 1) and high (Rentrop score of 2 or 3) coronary collateralization occurred in 161 and 242 patients, respectively. Serum levels of sVEGFR-1 and -2 were significantly elevated, in contrast, VEGF and PLGF levels were remarkably decreased in patients with low collateralization than in those with high collateralization (all P<0.05). Significant differences in sVEGFR-1, VEGF and PLGF levels was consistently detected between the low and high collateralization subgroups for patients with and without type 2 diabetes mellitus (DM) (for all comparisons, P<0.01). Multivariable regression analysis revealed that DM, dyslipidemia, elevated sVEGFR-1, and reduced VEGF and PLGF in serum were independently associated with a low degree of coronary collateralization. Increased serum sVEGFR-1 level is associated with poor coronary collateralization in patients with stable CAD. Type 2 DM is a predominant factor affecting collateral growth in these patients.

Changes of serum angiogenic biomarkers and their correlations with serum leptin concentration.

Obesity is considered as a major health problem. Angiogenic vessels by providing oxygen, nutrients and growth factors trigger growth and survival signals in adipocytes. We aimed to investigate the effect of high-fat diet (HFD) on serum angiogenic biomarkers including vascular endothelial growth factor (VEGF), soluble VEGF receptor 1 (sVEGFR1), nitric oxide (NO) concentrations and their correlations with serum leptin level in obese and control groups. Twenty male C57BL/6 mice were randomly assigned into the control and obese groups. Obese group received HFD for 15 weeks. At the end of experiment, blood samples were collected for blood glucose, serum insulin, VEGF, sVEGFR1, NO and leptin level measurements and correlation between serum angiogenic factors and leptin levels were analyzed. HFD induced higher serum NO and leptin levels compared to the control group, while, it did not affect serum VEGF and sVEGFR1 concentrations. There was a strong positive correlation between serum leptin and NO levels (r=0.78), however, a weak correlation was found between serum leptin and VEGF and VEGFR-1 concentrations. It seems that the angiogenic activities in obese mice are through the mechanisms that were not regulated by VEGF or VEGF receptors rather; other factors such as leptin and NO are involved (Tab. 1, Fig. 4, Ref. 32).