PlGF Levels Research Articles

Increased angiogenic factor secretion by decidual natural killer cells from pregnancies with high uterine artery resistance alters trophoblast function

STUDY QUESTIONAre the concentrations of factors secreted by decidual natural killer (dNK) cells from pregnancies at high risk of poor spiral artery remodelling different to those secreted from pregnancies at low risk?SUMMARY ANSWERExpression levels of PLGF, sIL-2R, endostatin and angiogenin were significantly increased by dNK cells from high-risk pregnancies, and angiogenin and endostatin were found to alter trophoblast function.WHAT IS KNOWN ALREADYDuring early pregnancy, maternal uterine spiral arteries are remodelled from small diameter, low-flow, high-resistance vessels into larger diameter, higher flow vessels, with low-resistance. This change is essential for the developing fetus to obtain sufficient oxygen and nutrients. dNK cells have been implicated in this process.STUDY DESIGN, SIZE, DURATIONdNK cells were isolated from first trimester terminations of pregnancies (obtained with local ethical approval) screened for normal- or high-resistance index, indicative of cases least (<1%) and most (>21%) likely to have developed pre-eclampsia had the pregnancy not been terminated (n = 18 each group). Secreted factors and the effects of these on the trophoblast cell line, SGHPL-4, were assessed in vitro.PARTICIPANTS/MATERIALS, SETTING, METHODSA multiplex assay was used to assess dNK cell-secreted factors. SGHPL-4 cell functions were assessed using time-lapse microscopy, 3D invasion assays, endothelial-like tube formation ability and western blot analysis.MAIN RESULTS AND THE ROLE OF CHANCEThe expression levels of PLGF (P < 0.01), sIL-2R (P < 0.01), endostatin (P < 0.05) and angiogenin (P < 0.05) were significantly increased by dNK cells from high-risk pregnancies. Endostatin significantly decreased SGHPL-4 invasion (P < 0.05), SGHPL-4 tube formation (P < 0.05) and SGHPL-4 Aktser473 phosphorylation (P < 0.05). Angiogenin significantly decreased SGHPL-4 invasion (P < 0.05), but increased SGHPL-4 tube formation (P < 0.01) and decreased SGHPL-4 Aktser473 phosphorylation (P < 0.05).LIMITATIONS, REASONS FOR CAUTIONThe culture of dNK cells and protein concentrations in vitro may not fully represent the in vivo situation. Although SGHPL-4 cells are extravillous trophoblast derived, further studies would be needed to confirm the roles of angiogenin and endostatin in vivo.WIDER IMPLICATIONS OF THE FINDINGSThe altered expression of secreted factors of dNK cells may contribute to pregnancy disorders associated with poor spiral artery remodelling.STUDY FUNDING/COMPETING INTEREST(S)This study was supported by the Wellcome Trust (project reference 091550). R.F. was a recipient of a PhD studentship from the Division of Biomedical Sciences, St. George's, University of London. The authors have no conflict of interests.

Biomarkers of endothelial dysfunction in preeclampsia and neonatal morbidity: a case–control study

ObjectiveTo investigate the association of preeclampsia with angiogenic imbalance, and the correlation of levels of angiogenic and anti-angiogenic factors to complications in mother and fetus. Study designSerum samples were obtained from 40 women with established preeclampsia (study group) and from 40 normotensive women (control group). Epidemiological characteristics of the two groups were analyzed. The levels of the angiogenic (VEGF and PlGF) and anti-angiogenic (sFlt-1) factors of the two study groups were determined in serum using ELISA. Neonatal adverse outcomes (late preterm, early term, low birth weight (LBW), very LBW (VLBW), intrauterine growth restriction (IUGR), and neonatal intensive care unit (NICU) admission) between the groups of the study were analyzed, as well as the association between the biomarkers of the study and neonatal adverse outcomes of the preeclamptic group of patients. ResultssFlt-1 levels were significantly higher in the preeclamptic women compared to normotensive women (median (range): 21297 (690–32637)pg/ml vs. 846.45 (363–2867)pg/ml, respectively), whereas there was a significant decrease in the levels of VEGF (90 (90–211)pg/ml vs. 90.55 (90–521)pg/ml, respectively), as well as in the levels of PlGF (13.62 (8–532)pg/ml vs. 239.86 (61–685)pg/ml, respectively). The increased serum values of the anti-angiogenic sFlt-1 were associated with increased rates of late preterm and early term births and VLBW. ConclusionAn imbalance between angiogenic and anti-angiogenic factors exists in preeclampsia and is associated with adverse maternal and neonatal outcomes.

Placental growth factor, pregnancy-associated plasma protein-A, soluble receptor for advanced glycation end products, extracellular newly identified receptor for receptor for advanced glycation end products binding protein and high mobility group box 1 levels in patients with acute kidney injury: a cross sectional study

BackgroundPlacental growth factor (PlGF), pregnancy-associated plasma protein-A (PAPP-A), soluble receptor for advanced glycation end products (sRAGE), extracellular newly identified receptor for RAGE binding protein (EN-RAGE) and high mobility group box 1 (HMGB-1) are novel biomarkers in chronic kidney disease (CKD). However, their clinical significance in acute kidney injury (AKI) is unknown. The aim of this cross-sectional study was to determine whether selected biomarkers are changed in AKI patients.MethodsSerum PlGF, PAPP-A, sRAGE, EN-RAGE and HMGB-1 levels were assessed in 40 patients with AKI, 42 CKD 5 patients, 31 haemodialysis patients (HD) and 39 age-matched healthy controls.ResultsPAPP-A was elevated in AKI (20.6 ± 16.9 mIU/L) compared with controls (9.1 ± 2.3 mIU/L, p < 0.001). PlGF was not increased in AKI (11.7 ± 7.4 pg/mL) versus controls (8.5 ± 2.4 pg/mL, n.s.), as well as sRAGE was not elevated in AKI (2400 ± 1400 pg/mL) compared with controls (1760 ± 730 pg/mL, n.s), but was lower compared with CKD 5 (3200 ± 1500 pg/mL, p < 0.05); EN-RAGE was elevated in AKI 480 ± 450 ng/mL in comparison with controls (60 ± 62 ng/mL), CKD 5 (190 ± 120 ng/mL), and HD (120 ± 100 ng/mL), all p < 0.001. Similarly, HMGB-1 was increased in AKI (5.8 ± 7.5 ng/mL) versus controls (1.7 ± 1.4 ng/mL), CKD 5 (3.2 ± 3.1 ng/mL) and HD (2.5 ±2.1 ng/mL), all p < 0.001.In AKI group, in multivariate regression analysis: PAPP–A levels were associated with transferrin (p <0.001), negatively with albumin (p < 0.01) and prealbumin (p < 0.05); PlGF levels were associated with C - reactive protein (p < 0.001). EN-RAGE levels were associated with ferritin (p < 0.01) and orosomucoid (p = 0.02), and HMGB-1 levels with leukocyte count (p < 0.01) and negatively with proteinuria (p = 0.02).ConclusionsIn AKI patients, PAPP-A, EN-RAGE and HMGB1 are elevated, but sRAGE and PlGF are not increased. Whereas PAPP-A correlates with markers of nutrition; PlGF, EN-RAGE and HMGB-1 are related to inflammatory parameters.

The Association of Alternate VEGF Ligands with Resistance to Anti-VEGF Therapy in Metastatic Colorectal Cancer

BackgroundCirculating angiogenic factors are altered in patients with mCRC receiving bevacizumab. Evaluation of alterations in levels of VEGF ligands may provide insights into possible resistance mechanisms.MethodsPlGF, VEGF-A, VEGF-C, and VEGF-D were measured from two cohorts of patients. Sequential plasma samples were obtained from a discovery cohort of 42 patients treated with chemotherapy and bevacizumab. A validation cohort included plasma samples from a cross-sectional of 403 patients prior to chemotherapy, or after progression on a regimen with or without bevacizumab.ResultsIn the discovery cohort, VEGF-C was increased prior to progression and at progression (+49% and +95%, respectively, p<0.01), consistent with previously reported elevations in PlGF. Levels of VEGF-D were increased (+23%) at progression (p=0.05). In the validation cohort, samples obtained from patients after progression on a regimen with bevacizumab had higher levels of PlGF and VEGF-D (+43% and +6%, p=0.02, p=0.01, respectively) compared to untreated patients, but failed to validate the increase in VEGF-C seen in the first cohort. Patients who progressed on chemotherapy with bevacizumab had significantly elevated levels of PlGF (+88%) but not VEGF-C and VEGF-D compared to patients treated with chemotherapy alone. Elevations of PlGF and VEGF-D appeared transient and returned to baseline with a half-life of 6 weeks.ConclusionsIncreases in PlGF and VEGF-D were observed after progression on chemotherapy with bevacizumab. These changes appear to be reversible after discontinuing therapy. These ligands are associated with resistance to bevacizumab-containing chemotherapy in mCRC, but causation remains to be established.

Blood outgrowth endothelial cells in patients with severe ischemic cardiomyopathy maintain pro-angiogenic phenotype without senescence

Purpose: Clinical translation of cardiovascular cell therapy with proven benefits in experimental models may be hampered by a dysfunctional progenitor cell phenotype in heart failure patients, as reported for bone marrow mononuclear cells. We compared blood outgrowth endothelial cells (BOECs) from patients with severe but stable ischemic heart disease (IHD) and healthy controls (CON). Methods: BOECs were derived from 37 IHD (LVEF≤45%) and 35 CON. Cell surface markers were analysed by flow cytometry to confirm BOEC-phenotype. In vitro functional analysis, including tube formation potential (on matrigel), senescence (SA-β-gal) and Dil-AcLDL uptake (flow cytometry) was performed between passage 3-5 and expression of angiogenic genes was studied using quantitative RT-PCR. Results: An average of 3.4±0.5 BOEC-colony forming units (CFUs)/100×106 mononuclear cells (MNCs) was obtained in IHD (age: 66±2y, LVEF: 31±2%, LVEDD: 56±1mm) vs 2.5±0.4 CFUs/100×106 MNCs in CON (age: 36±2y) (P=0.12). In patients, BOEC-outgrowth did not correlate with the extent of LV systolic dysfunction (R2=0.03) or LV remodeling (R2=0.05), and was not affected by cardiovascular risk factors. A mild correlation was observed between age and CFU number (R2=0.13, P=0.04). Endothelial lineage markers (CD309/CD31/CD146) were comparable in IHD and CON and panleukocyte marker CD45 was absent in all colonies. Progenitor marker CD133 was absent, while CD34 displayed important inter-sample variability and tended to be more expressed in CON (77±10%, n=9) vs IHD (54±8%, n=13) (P=0.06). Dil-AcLDL uptake was more pronounced in BOECs from IHD (98±1%, n=13) vs CON (94±2%, n=9) (P=0.008). In vitro tube formation, total tube length, and number of intersections were comparable in IHD and CON (n=13 vs 9, resp). The percentage senescent BOECs was similar in IHD and CON (7±2%, n=14 vs 8±3%, n=9). Finally, robust expression levels of PLGF, Angiopoietin-2, PDGF-β, FGF-2 and VEGF were comparable in IHD (n=15) and CON (n=9). Conclusions: BOECs can be successfully and consistently derived from patients with IHD and have a preserved angiogenic profile. Therefore, BOECs hold promise for clinical translation of autologous cell therapy in severe ischemic cardiomyopathy.

Analysis of serum biomarkers and tumor genetic alterations from a phase II study of lenvatinib in patients with advanced BRAF wild-type melanoma.

9058 Background: Lenvatinib is an oral receptor tyrosine kinase inhibitor targeting VEGFR1-3, FGFR1-4, RET, KIT, and PDGFRβ. Phase I and II studies of lenvatinib demonstrate that a subset of advanced melanoma patients (pts) appears to derive benefit from lenvatinib treatment (tx), prompting a need to identify predictive biomarkers for response to lenvatinib. Methods: Serum and archival tumor samples were collected from 93 enrolled pts in the BRAF wild-type (wt) cohort of the lenvatinib phase II study of advanced melanoma. Concentrations of 50 factors were measured in pre- and post-tx serum samples; 58 archival tumor tissues were obtained and gene mutation (mut) (n=53) and gene expression profiling (GEP) (n=39) analyses were performed. For mut analysis, targeted resequencing was performed on a select panel of genes using the Ion PGM Sequencer and mut validation studies were performed. For GEP analysis, the nCounterAnalysis System was used to measure the expression level of 330 genes. Results: Clinical benefit rate (32%) and objective response rate (9%) by independent radiologic review were observed in the BRAF wt cohort. In serum biomarker analysis, lenvatinib tx resulted in a decrease in soluble VEGFR2 and increase in VEGF and PlGF levels consistent with target engagement. Correlation with clinical outcome demonstrated that baseline (BL) levels of Ang-2, IL8, PlGF, and PDGFBB associated with OS, but only pts with low BL Ang-2 levels also demonstrated improved response. High BL levels of FLT3LG and eotaxin associated with longer OS and improved response. In GEP analysis, expression levels of a set of genes including TARBP2, ZNF544, and NF1 (targets identified in phase I and preclinical studies) associated with OS. In gene mut analysis, NRAS mut showed a trend towards longer OS. Pts with NRAS mut along with wt PIK3CA status demonstrated longer OS. Conclusions: Lenvatinib demonstrated limited response in BRAF wt advanced melanoma pts. A subset of pts may derive extended clinical benefit. NRAS mut/PIK3CA wt status and low BL Ang-2 levels appear to associate with improved clinical outcome in this pt population and require further study to assess their predictive value. Clinical trial information: NCT01136967.

Circulating oncometabolite 2-hydroxyglutarate (2HG) as a potential surrogate biomarker in patients with isocitrate dehydrogenase mutant (IDHm) intrahepatic cholangiocarcinoma (ICC).

4125 Background: Mutations in the genes encoding for IDH1 and IDH2 occur in ~20% of ICC patients (pts), and they lead to the production of the oncometabolite 2HG. We examined whether serum 2HG levels in IDHm ICC pts may 1) serve as a surrogate biomarker for IDH status, 2) correlate with tumor burden, and 3) correlate with circulating proangiogenic biomarkers. Methods: Blood samples from 33 ICC pts [11 IDHm, 22 IDH wild-type (IDHwt)] of different AJCC stages from MGH and 39 surgically resected ICC patients (7 IDHm, 32 IDHwt) from HKU were analyzed for serum 2HG concentration by reverse-phase liquid chromatography coupled to mass spectrometry. Eight circulating proangiogenic biomarkers were measured in plasma using multiplex ELISA. Results: In the MGH cohort, median serum 2HG levels were significantly elevated in IDHm (478 ng/ml [interquartile range 174–643]) versus IDHwt ICC pts (118ng/ml [68–160])(p&lt;0.001). Similarly, in the HKU cohort, the pre-resection median serum 2HG levels were significantly elevated in IDHm (343ng/ml [192–596]) versus IDHwt ICC pts (56ng/ml [42–81]) (p&lt;0.0001). The area under ROC curve for prediction of an IDH mutation using 2HG was 93%; with a threshold of 2HG≥170ng/ml, the sensitivity was 83% and specificity was 90%. IDH2 mutations were more frequent in the HKU cohort (3/7, 43%) compared with the MGH cohort (0/11, 0%) (p&lt;0.05), but 2HG levels did not differ among the specific IDH1 or IDH2 allelic variants. 2HG levels correlated directly with tumor burden (Spearman’s rho=0.89; p&lt;0.05) in the HKU cohort. Median plasma levels of PlGF—a growth factor from the VEGF-family—were higher in IDHm (35pg/ml [33–40]) versus IDHwt ICC pts (median 26pg/ml [24–34]) from the HKU cohort (p&lt;0.05). No other associations were seen between proangiogenic biomarkers and IDH status. Conclusions: IDHm ICC pts had significantly higher serum 2HG levels compared to IDHwt ICC pts. High serum 2HG correlated with increased tumor burden. Pre-resection circulating PlGF levels were higher in IDHm ICC versus IDHwt pts. These data support further exploration of circulating 2HG as potential surrogate and response biomarker in IDHm ICC.

PP052. Diagnosis of pre-eclampsia and prediction of time to delivery in women with suspected pre-eclampsia presenting before 36 weeks’ gestation

The pro-angiogenic factor PlGF has an important role in the development and maintenance of the placenta. Incomplete remodelling of uterine spiral arteries is associated with placental oxidative stress, placental dysfunction, and the complications of pre-eclampsia and fetal growth restriction. Abnormally low PlGF levels identify placental dysfunction and an increased risk for pregnancy complication. To evaluate the correlation between abnormally low PlGF concentration, a diagnosis of pre-eclampsia or IUGR, and a short interval to delivery. We recruited a total of 45 pregnant women with suspected pre-eclampsia between 23+0 to 36+0 weeks of gestation. The concentration of PlGF was measured in maternal EDTA plasma samples (Alere, Triage® PlGF), a rapid assay using monoclonal antibodies raised against free PlGF. Pre-eclampsia was diagnosed clinically in 25 women. PlGF was abnormal (<100pg/mL) in all 25 women (100% sensitivity). Eighteen women (72%) had a PlGF level <12pg/mL (very abnormal) and, of these, 14 (78%) had IUGR and a birthweight below the 10th centile. The mean time to delivery in women with a PlGF level <12pg/mL was 7.5 days (0-18d), compared with 23 days (0-67d) for a PlGF level in the range ⩾12<100pg/mL, in agreement with data recently reported by Chappell [1]. The high sensitivity of PlGF for pre-eclampsia with/without IUGR and its strong prediction of time-to-delivery allows PlGF to be a valuable diagnostic tool. PlGF has the potential to remove the uncertainty in clinical decision-making in women with suspected pre-eclampsia.

PP051. Budget impact analysis of maternal plasma PIGF concentrations in women with suspected pre-eclampsia: The potential for improved health service usage

Existing methods of assessing women with suspected pre-eclampsia are expensive and labour intensive, yet perform poorly. The PELICAN multi-centre observational cohort study demonstrated that PlGF predicts need for delivery for pre-eclampsia within 14 days. We constructed a decision analytical model using outcome data from 100 women comparing resource use by final diagnosis and PlGF level. Costs were obtained from 2012-2013 NHS tariffs. Of the 100 women, 40 had a final diagnosis of pre-eclampsia and delivered within 14 days of the PlGF test; 10 were healthy pregnancies with normal PlGF levels. 82% of women with pre-eclampsia were admitted during the final 2 weeks of pregnancy, with a 5-day (SD=5) length of stay, compared with 30% of healthy pregnancies with normal PlGF (1 day; SD=0.58). Resource use for outpatient appointments, scans and day unit admissions was similar for the two groups although higher in the group with a final diagnosis of pre-eclampsia. Total costs, excluding delivery, were approximately a third higher in the pre-eclampsia group. This interim analysis suggests pre-eclampsia is associated with significantly higher resource use, although there is some inappropriate resource use in healthy women. PlGF can assist diagnosis and identify women requiring increased care and could be used to direct appropriate resource allocation in women over 37 weeks.

PP037. Relationship of serum leptin levels to circulating cytokines, chemokines, adhesion molecules and angiogenic factors in women with preeclampsia

Serum leptin levels are known to be increased in preeclampsia. To determine circulating levels of leptin along with those of several cytokines, chemokines, adhesion molecules and angiogenic factors in normal pregnancy and preeclampsia, and to investigate whether serum leptin levels are related to the clinical features and measured laboratory parameters of the patients. Serum levels of leptin, IL-1beta, IL-1ra, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p40, IL-12p70, IL-18, IFN-gamma, TNF-alpha, TGF-beta1, IP-10, MCP-1, ICAM-1, VCAM-1, sFlt-1 and PlGF were determined in 60 preeclamptic, 60 healthy pregnant and 59 healthy non-pregnant women. There were significant differences in most of the measured laboratory parameters among the three study groups except for serum IL-1beta and TGF-beta1 levels. Preeclamptic patients had significantly higher leptin levels than healthy pregnant women. A significant positive correlation was observed between serum leptin and IP-10 concentrations of preeclamptic patients. Furthermore, elevated serum leptin level and sFlt-1/PlGF ratio had an additive effect in the risk of preeclampsia, as shown by the substantially higher odds ratios of their combination than of either alone. Increased serum levels of leptin are related to those of IP-10 in preeclampsia, suggesting that circulating leptin might contribute to the development of endothelial dysfunction through the anti-angiogenic effect of IP-10.

Efficacy, Safety, Pharmacokinetics, and Biomarkers of Cediranib Monotherapy in Advanced Hepatocellular Carcinoma: A Phase II Study

We conducted a single-arm phase II study of cediranib, a pan-VEGFR tyrosine kinase inhibitor, in patients with advanced hepatocellular carcinoma (HCC). Patients with histologically confirmed measurable advanced HCC and adequate hematologic, hepatic, and renal functions received cediranib 30-mg orally once daily (4 weeks/cycle). The primary endpoint was progression-free survival (PFS) rate at 3 months. Other endpoints included response rates, overall survival (OS), pharmacokinetics (PK), and biomarkers for cediranib. Cediranib treatment resulted in an estimated 3-month PFS rate of 77% (60%, 99%). Median PFS was 5.3 (3.5,9.7) months, stable disease was seen in 5/17 patients (29%), and median OS was 11.7 (7.5-13.6) months. Grade 3 toxicities included hypertension (29%), hyponatremia (29%), and hyperbilirubinemia (18%). Cediranib PK were comparable to those seen in cancer patients with normal hepatic function. Plasma levels of VEGF and PlGF increased and sVEGFR1, sVEGFR2, and Ang-2 decreased after cediranib treatment. PFS was inversely correlated with baseline levels of VEGF, sVEGFR2, and bFGF and with on-treatment levels of bFGF and IGF-1, and directly associated with on-treatment levels of IFN-γ. OS was inversely correlated with baseline levels of sVEGFR1, Ang-2, TNF-α, CAIX, and CD34(+)CD133(+)CD45(dim) circulating progenitor cells and on-treatment levels of sVEGFR2. Despite the limitations of primary endpoint selection, cediranib at 30-mg daily showed a high incidence of toxicity and preliminary evidence of antitumor activity in advanced HCC. Hepatic dysfunction did not seem to affect the steady-state PK of cediranib. Exploratory studies suggested proangiogenic and inflammatory factors as potential biomarkers of anti-VEGF therapy in HCC.

Gestation Dependant Changes in Angiogenic Factors and Their Associations with Fetal Growth Measures in Normotensive Pregnancy

BackgroundEarlier studies indicate that altered angiogenesis at birth is associated with poor birth outcome in women with preeclampsia. Now, we hypothesize that the progressive gestation dependant changes in markers of angiogenesis will be more useful to predict birth weight early even in a normotensive pregnancy. This study for the first time examines the association of gestation dependant changes in the levels of maternal angiogenic factors in addition to their levels in cord with birth weight.MethodNinety two pregnant women were followed at three different time points: 16–20 weeks, 26–30 weeks and at delivery during pregnancy. Plasma levels of angiogenic and anti angiogenic factors were determined by commercial enzyme-linked immunosorbent assay (ELISA) kits.ResultsMaternal plasma VEGF levels increased (p<0.01) till the second time point and decreased (p<0.05) up to delivery while plasma sFlt-1 levels increased (p<0.01) at delivery. PlGF levels peaked (p<0.01) at second time point and decreased (p<0.01) at delivery. Cord plasma VEGF levels were higher (p<0.01) and sFlt-1 levels were lower (p<0.01) as compared to maternal values at all time points. Maternal plasma VEGF levels at first time point and PlGF levels at delivery were positively (p<0.05 and p<0.01 respectively), while sFlt-1/PlGF ratio at delivery was negatively associated (p<0.05) with birth weight.ConclusionLevels of pro- and anti-angiogenic factors may be differentially regulated across gestation. Maternal VEGF levels at early gestation (16–20 weeks) may be predictive of birth weight in healthy term pregnancies.

An Elevated Maternal Plasma Soluble fms-Like Tyrosine Kinase-1 to Placental Growth Factor Ratio at Midtrimester Is a Useful Predictor for Preeclampsia

Background. To assess the ability of mid-trimester sFlt-1/PlGF ratio for prediction of preeclampsia in two different Arabic populations. Methods. This study measured levels of sFlt-1, PlGF, and sFlt-1/PlGF ratio at midtrimester in 83 patients who developed preeclampsia with contemporary 250 matched controls. Results. Women subsequently developed preeclampsia had significantly lower PlGF levels and higher sFlt-1 and sFlt-1/PlGF ratio levels than women with normal pregnancies (P < 0.0001 for all). Women who with preterm preeclampsia had significantly higher sFlt-1 and sFlt-1/PlGF ratio than term preeclamptic women (P = 0.01, 0.003, resp.). A cutoff value of 3198 pg/mL for sFlt-1 was able to predict preeclampsia with sensitivity, specificity, and accuracy of 88%, 83.6%, and 84.7%, respectively, with odds ratio (OR) 37.2 [95% confidence interval (CI) 17.7–78.1]. PIGF at cutoff value of 138 pg/mL was able to predict preeclampsia with sensitivity, specificity, and accuracy of 85.5%, 77.2%, and 79.3%, respectively, with OR 20 [95% CI, 10.2–39.5]. The sFlt-1/PIGF ratio at cutoff value of 24.5 was able to predict preeclampsia with sensitivity, specificity, and accuracy of 91.6%, 86.4%, and 87.7%, respectively with OR 67 [95% CI, 29.3–162.1]. Conclusion. Midtrimester sFlt-1/PlGF ratio displayed the highest sensitivity, specificity, accuracy, and OR for prediction of preeclampsia, demonstrating that it may stipulate more effective prediction of preeclampsia development than individual factor assay.

648: The impact of change in pregnancy body mass index on gestational hypertension/preeclampsia

gestational hypertension/preeclampsia Morgan Swank, Aaron Caughey, Christine Farinelli, Elliott Main, Kathryn Melsop, William Gilbert, Judith Chung University of California, Irvine, Maternal Fetal Medicine, Orange, CA, Oregon Health & Science University, Maternal Fetal Medicine, Portland, OR, California Maternal Quality Care Collaborative, CMQCC, Stanford, CA, Sutter Medical Center, Maternal Fetal Medicine, Sacramento, CA OBJECTIVE: To examine the impact of changes in body mass index (BMI) during pregnancy on the incidence of gestational hypertension (GHTN)/preeclampsia (PRE). STUDY DESIGN: This is a retrospective cohort study using linked birth certificate and discharge diagnosis data (All-California, Rapid-Cycle, Maternal/Infant Database) from the year 2007. Adjusted odds ratios (aOR) with 95% confidence intervals (CI) were calculated for the outcome of GHTN/PRE, as a function of a categorical change in pregnancy BMI: BMI loss (BMI change 0.5), no change ( 0.5 to 0.5), minimal (0.6 to 5), moderate (5.1 to 10), and excessive ( 10). No change in BMI served as the reference group. The impact of pregnancy change in BMI was determined for the entire cohort and then stratified by prepregnancy WHO BMI category. RESULTS: The study population consisted of 436,414 women with singleton gestations. Overall, women with excessive BMI change had a nearly two-fold increased incidence of GHTN/PRE, when compared to women with no net change in BMI (aOR 1.94; 95% CI 1.722.20). By prepregnancy BMI class, the aOR for GHTN/PRE in women with excessive BMI change were: normal weight (aOR 2.74, 95% CI, 1.87-4.04), overweight (aOR 3.43, 95% CI 2.62-4.94), obese class I (aOR 2.47, 95% CI 1.93-3.18), obese class II (aOR 2.53, 95% CI 1.84-3.48), obese class III (aOR 2.89, 95% CI 2.00-4.19). CONCLUSION: Regardless of prepregnancy BMI category, the aOR of developing GHTN/PRE was significantly higher in women with BMI change 10 when compared to women with no or minimal BMI change. Women who entered pregnancy in the overweight category (BMI 25.0-29.9) were at the greatest risk for developing hypertensive disorders with excessive BMI change. This may provide beneficial material for counseling patients of normal and overweight status, who are typically considered to be of a lower risk strata. Unadjusted incidence of gestational hypertension/preeclampsia as a function of change in pregnancy body mass index 649 PTX3, sFlt-1 and PlGF levels in mothers and their own newborn Paola Algeri, Sara Ornaghi, Davide Bernasconi, Fabrizio Cappellini, Stefano Signorini, Paolo Brambilla, Gabriele Urban, Patrizia Vergani University of Milan-Bicocca, Obstetrics and Gynaecology, Monza, Italy, University of Milan-Bicocca, Clinical medicine and Prevention, Center of Biostatistic for Clinical Epidemiology, Monza, Italy, University of MilanBicocca, Laboratory Medicine, Desio, Italy, University of Milan-Bicocca, Experimental Medicine, School of Medicine, Milan, Italy, University of Milan-Bicocca, Obstetrics and Gynaecology, Desio, Italy OBJECTIVE: An alteration of PTX3, sFlt-1 and PlGF serum levels is described during preeclampsia. Instead, no data about these markers in fetal serum are reported in literature. The aim of our study is to compare serum maternal and fetal markers’ levels in order to evaluate a potential relation between them. STUDY DESIGN: A case-control study with collection of 74 maternal serum samples and 74 fetal serum samples obtained from umbilical blood of each own baby. Samples were collected at time of delivery in 22 preeclamptic women and their newborns (cases), 23 women and their own babies, who formed a gestational age homogeneous group compared to cases (including pregnancies complicated by preterm delivery, IUGR and pPROM), and 29 healthy controls and their newborns (normal pregnancies delivered by elective cesarean section at 37 weeks). Statistical analysis was performed with Paired Wilcoxon test and Spearman correlation. RESULTS: Maternal and fetal levels of PTX3, sFlt-1 and PlGF were found to be different (p 0.0001 for each comparison). For each marker, the fetal level median was lower than the maternal one (PTX3: maternal median 1.78 ng/ml vs fetal median 1.11; sFlt-1: maternal median 5001.5 pg/ml vs fetal one 242.6; PlGF: maternal median 94.3 pg/ml vs fetal one 11.2).Table 1 shows Spearman correlation results: an independent trend for PTX3 in maternal versus fetal serum (p 0.14) was found, whereas maternal and fetal levels of sFlt-1 and PlGF presented a directly proportional relationship. CONCLUSION: Our results support a possible independent production of PTX3 in the fetus, whereas a trans-placental passage from mother to fetus is more likely to be the mechanism underlying fetal sFlt-1 and Demographics and miRNA levels between cases and controls

650: sTRAIL, a prognostic marker for future death from cardiovascular disease, is decreased in maternal plasma of patients with preeclampsia

650 sTRAIL, a prognostic marker for future death from cardiovascular disease, is decreased in maternal plasma of patients with preeclampsia Piya Chaemsaithong, Tinnakorn Chaiworapongsa, Tamara Stampalija, Nandor Gabor Than, Zhong Dong, Lami Yeo, Sonia Hassan, Roberto Romero NICHD/NIH/DHHS, Perinatology Research Branch, Detroit, MI, Wayne State University School of Medicine, Department of Obstetrics and Gynecology, Detroit, MI OBJECTIVE: Women who develop preeclampsia (PE) are at increased risk of cardiovascular disease later in life. Tumor necrosis factor (TNF)-Related Apoptosis-Inducing Ligand (TRAIL), a potent inducer of apoptosis, has anti-atherosclerotic effects on endothelial cells. Patients with acute coronary artery disease have lower serum concentrations of the soluble TRAIL (sTRAIL), with higher C-reactive protein (CRP) concentrations. sTRAIL and CRP confer a risk for future death (Hazard Ratio 0.93 and 2.2, respectively). The aim of this study was to determine if maternal plasma concentrations of sTRAIL and CRP change in pregnancies with preeclampsia. STUDY DESIGN: A cross-sectional study was conducted in the following groups: 1) normal pregnant women (n 93); and 2) women with PE (n 52). Patients with PE were sub-classified into preterm (n 38) and term ( 37 weeks) PE (n 14). Maternal plasma concentrations of sTRAIL and C-reactive protein (CRP) were determined by ELISA. RESULTS: 1) The median plasma concentration of sTRAIL was significantly lower in patients with PE than in normal pregnant women (25.6 pg/mL vs. 29.2 pg/mL; p 0.03); 2) the preterm (but not term) preeclampsia group had a significantly lower median plasma concentration of sTRAIL than the normal pregnancy group (24.8 pg/mL vs. 30.5 pg/mL; p 0.03 and 26.5 pg/mL vs. 26.8 pg/mL; p 0.7, respectively); and 3) the median plasma concentration of CRP was significantly higher in patients with PE (n 32) than in normal pregnant women (n 53) (8 ng/mL vs. 4.1 ng/mL; p 0.001); however, there was no significant correlation between maternal plasma concentrations of sTRAIL and CRP in either PE (p 0.1) or normal pregnancy (p 0.3). CONCLUSION: Preeclampsia was associated with a lower sTRAIL and a higher CRP concentration in maternal plasma. Both proteins may be involved in the mechanisms of predisposing patients with preeclampsia to future cardiovascular disease. 651 Prospective evaluation of circulating angiogenic factors in the prediction of earlyversus late-onset pre-eclampsia Rebecca Jessel, Ann Thomas, Kee-Hak Lim, Samuel Parry, Thomas McElrath Brigham and Women’s Hospital; Harvard Medical School, Department of Obstetrics and Gynecology, Boston, MA, Brigham and Women’s Hospital; Harvard Medical School, Department of Obstetrics and Gynecology; Division of Maternal Fetal Medicine, Boston, MA, Boston MFM; BIDMC; Harvard Medical School, Boston Maternal-Fetal Medicine, Boston, MA, Hospital of the University of Pennsylvania, Division of Maternal-Fetal Medicine, Hospital of the University of Pennsylvania, PA OBJECTIVE: Circulating angiogenic factors play a role in pathogenesis of preeclampsia (PE) but we have previously demonstrated that they have limited predictive capacity if PE is considered a single entity. However, PE is a heterogeneous disease with early ( 34w) and late onest PE possibly representing different entities. We tested the hypothesis that sequential measurement of sFlt-1 and PlGF in a longitudinal cohort of patients can distinguish patients who are more likely to develop and require delivery for early PE. STUDY DESIGN: 2,241 singleton pregnancies were followed prospectively from the initiation of prenatal care at three regional academic centers. Plasma sFlt-1 and PlGF levels were collected prospectively and quantified by immunoassay at approximately 10, 17, and 24 weeks gestation. Patients were followed through delivery. PE was diagnosed by ACOG criteria. Patients with chronic hypertension and diabetes were excluded from this analysis. RESULTS: Median analyte concentrations for early and late and testing characteristics for the prediction of early PE are presented in the table. Twenty patients developed PE before 34 weeks compared to 182 patients with late PE. Incidence of early PE was 1%. PlGF concentrations are significantly lower among early PE patients at all time points and magnitude of increase with gestation was lower than those with late PE. sFlt-1 concentrations and the ratio are significantly different at 26 weeks. However, the testing characteristics do not suggest clinical utility in the prediction of early PE. CONCLUSION: Our results support the suggestion that PE is a heterogeneous disease with early and late PE possibly representing different disease processes. Despite low incidence, the significant differences in analytes in early versus late suggests a more homogenous cohort. However, the analytes, do not appear to be clinically useful in the prediction of early PE.

649: PTX3, sFlt-1 and PlGF levels in mothers and their own newborn

649: PTX3, sFlt-1 and PlGF levels in mothers and their own newborn

634: DNA methylation patterns of first trimester trophoblast in severe preeclampsia

vs. 6.2, p 0.033). PlGF and sFlt-1 levels were not significantly different between women with late-onset PE (delivery 36 weeks) and controls without PE. CONCLUSION: Higher plasma PlGF prior to the establishment of hemochorial circulation and lower after in early PE suggests a possible response of the nascent placenta to aberrant placentation and an altered hypoxic environment. The pattern of changes in sFlt-1 levels with hemochorial circulation in patients with early PE may represent a similar response. Taken together, these results confirm that the conditions of PE are established in the first trimester.

633: Maternal plasma levels of PlGF and sFlt-1 change throughout the 1st trimester and correlate with development of early-onset pre-eclampsia

633: Maternal plasma levels of PlGF and sFlt-1 change throughout the 1st trimester and correlate with development of early-onset pre-eclampsia

Placental Growth Factor for the Prediction of Adverse Outcomes in Patients with Suspected Preeclampsia or Intrauterine Growth Restriction

BackgroundThe circulating concentration of PlGF is reported to be lower in patients experiencing preeclampsia and patients delivering a small for gestational age (SGA) neonate. To evaluate the predictive value of circulating PlGF for preeclampsia and adverse outcome in patients with suspected preeclampsia or intrauterine growth restriction (IUGR).Methodology/Principal FindingsA double blind prospective study. We enrolled 96 women for suspected preeclampsia or IUGR, and measured plasma levels of PlGF (Triage®) at enrolment. We defined adverse outcome as severe preeclampsia, SGA neonate (<10th centile) or elective delivery for maternal or fetal complication. Severe adverse outcome was studied among patients included <34 weeks gestation (WG) and defined as eclampsia, HELLP syndrome, very SGA (<3rd centile) or elective delivery <34 WG. The mean logtransformed PlGF level was lower for women who experienced preeclampsia (2.9 vs 3.7, p = 0.02), and was markedly lower for patients who experienced adverse outcome (2.9 vs 4.3, p<0.001). The odds of presenting an adverse outcome were higher for the lowest tertile of PlGF compared to the higher (OR = 13 , 95% CI [3–50]). For severe adverse outcome, odds were respectively for the lowest and intermediate tertile as compared with the higher tertile : OR = 216, 95% CI [18–2571]; and OR = 17, 95% CI [3–94]. When included <34 WG, patients with a PlGF level <12 pg/ml experienced a severe adverse outcome in 96% of cases (24/25), and only 1 of 20 patients with a PlGF level >5th centile experienced a severe adverse outcome within 15 days (5%).Conclusions/SignificanceAmong women with suspected preeclampsia or IUGR, PlGF helps identify women who will experience an adverse outcome and those who will not within a time period of 15 days.

Serum sFlt-1 and PlGF levels can be used to distinguish between CKD and pre-eclampsia in pregnant women

Serum sFlt-1 and PlGF levels can be used to distinguish between CKD and pre-eclampsia in pregnant women