sFlt-1 Levels Research Articles

The role of vascular inflammation markers in deficiency of adenosine deaminase 2

ObjectivesThe first objective was to assess the role of vascular inflammatory factors in the pathogenesis of deficiency of adenosine deaminase 2 (DADA2) and to compare these markers among DADA2 patients with different phenotypes. We also aimed to investigate differences between DADA2 patients with vasculitic features and classic polyarteritis nodosa (PAN) for the aforementioned markers. MethodsThe study included eighteen DADA2 patients, ten PAN patients, and eight healthy controls. Plasma levels of sST2, sRAGE, Tie-2, sCD40L, Tie-1, sFlt-1, LIGHT, TNF-α, PlGF, IL-6, IL-18, IL-10, MCP-1 were studied by cytometric bead-based multiplex assay panel. ResultsAmong the DADA2 patients, five had hematological manifestations, 13 had vasculitic findings, and accompanying immunological findings were present in seven patients. Nine patients had neurological findings, five of whom had neuropathy. Plasma levels of Tie-1 and sFlt-1 were higher in the overall DADA2 patients compared to healthy controls and PAN patients (p<0.001 and p = 0.004, respectively). DADA2 patients with PAN-like features had higher sRAGE, Tie-2, and TNF-α levels compared to PAN patients (p = 0.013, p = 0.003, and p = 0.001, respectively).In DADA2 patients with hematological findings, plasma IL-18 levels were higher than those with PAN-like phenotype (p = 0.001). Finally, DADA2 patients with neuropathy had higher sRAGE concentrations than patients without neuropathy and healthy controls (p = 0.03 and p = 0.008, respectively). ConclusionsWe suggest that the high plasma IL-18 levels observed in DADA2 patients with hematologic manifestations may be associated with an activated IFNγ pathway, and lack of response to anti-TNF treatment. We identified sRAGE as a potential biomarker of neuropathy in DADA2 patients. Higher concentrations of Tie-1, Tie-2, sFlt-1, sRAGE, and TNF-α distinguished DADA2 patients with PAN-like features from PAN patients.

The sFlt-1 to PlGF ratio in pregnant women with rheumatoid arthritis: impact of disease activity and sulfasalazine use.

An elevated sFlt-1/PlGF ratio has been validated as a significant predictor of preeclampsia, but has not been established in women with RA. We explored whether the sFlt-1/PlGFratio could be altered due to disease activity in RA, and could be applied in this population to predict preeclampsia. Since SSZ has been suggested to improve the angiogenic imbalance in preeclampsia, we also aimed to examine whether SSZ could affect sFlt-1 or PlGF levels. Making use of a nationwide, observational, prospective cohort study on pregnant women with RA, sFlt-1 and PlGF were measured in the third trimester. A total of 221 women, aged 21-42 years, were included, with a median gestational age of 30 + 3 weeks. No differences in sFlt-1 or PlGF were observed between women with high, intermediate or low disease activity (P = 0.07 and P = 0.41), whereas sFlt-1 and PlGF did not correlate with DAS28-CRP score (r = -0.01 and r = -0.05, respectively). Four (2%) women with a sFlt-1/PlGF ratio ≤38 developed preeclampsia in comparison to three (43%) women with a ratio > 38, corresponding to a negative predictive value of 98.1%. SSZ users (n = 57) did not show altered levels of sFlt-1 or PlGF in comparison to non-SSZ users (n = 164, P = 0.91 and P = 0.11). Our study shows that in pregnant women with RA, the sFlt-1/PlGF ratio is not altered due to disease activity and a cut-off ≤38 can be used to exclude preeclampsia. Additionally, SSZ use did not affect sFlt-1 or PlGF levels in this population.

SARS-CoV-2 colonization of maternal and fetal cells of the human placenta promotes alteration of local renin-angiotensin system

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection appears to increase the risk of adverse pregnancy outcomes, such as pre-eclampsia in pregnant women. The mechanism(s) by which this occurs remains unclear.MethodsWe investigated the pathophysiology of SARS-CoV-2 at maternal-fetal interface in pregnant women who tested positive for the virus using RNA in situ hybridization (viral RNA), immunohistochemistry, and hematoxylin and eosin staining. To investigate whether viral infection alters the renin angiotensin system (RAS) in placenta, which controls blood pressure, we treated human trophoblasts with recombinant spike protein or a live modified virus with a vesicular stomatitis viral backbone expressing spike protein (VSV-S).FindingsViral colonization was highest in maternal decidua, fetal trophoblasts, Hofbauer cells, and in placentas delivered prematurely. We localized SARS-CoV-2 to cells expressing angiotensin-converting enzyme 2 (ACE2) and demonstrate that infected placentas had significantly reduced ACE2. In response to both spike protein and VSV-S, cellular ACE2 decreased although angiotensin II receptor type 1 (AT1R) increased with concomitant increase in soluble fms-like tyrosine kinase-1 (sFlt1). Viral infection decreased pro-angiogenic factors, AT2R, and placental growth factor, which competitively binds to sFlt1. Sera from infected pregnant women had elevated levels of sFlt1 and angiotensin II type 1-receptor autoantibodies prior to delivery, both signatory markers of pre-eclampsia.ConclusionsSARS-CoV-2 colonizes ACE2-expressing maternal and fetal cells in the placenta. Infection in pregnant women correlates with alteration of placental RAS. As RAS regulates blood pressure, SARS-CoV-2 infection may thus increase adverse hemodynamic outcomes, such as pre-eclampsia in pregnant women.FundingNIH/NICHD grants R01 HD091218 and 3R01HD091218-04S1 (RADx-UP Supplement).

Can placental growth factors explain birthweight variation in offspring of women with type 1 diabetes?

Maternal hyperglycaemia alone does not explain the incidence of large offspring amongst women with type 1 diabetes. The objective of the study was to determine if there is an association between placental function, as measured by angiogenic factors, and offspring birthweight z score in women with type 1 diabetes. This cohort study included samples from 157 Continuous Glucose Monitoring in Pregnant Women with Type 1 Diabetes (CONCEPTT) trial participants. Correlations were estimated between birthweight z score and placental growth factor (PlGF) and soluble fms-like tyrosine kinase (sFlt-1) levels measured at baseline and at 24 and 34weeks of gestation. Linear regression was used to assess the relationship between birthweight z score and placental health, as measured by PlGF and sFlt-1/PlGF ratio, stratified by glycaemic status (continuous glucose monitoring and HbA1c measures) and adjusted for potential confounders of maternal BMI, smoking and weight gain. Higher PlGF levels and lower sFlt-1/PlGF ratios represent healthy placentas, while lower PlGF levels and higher sFlt-1/PlGF ratios represent unhealthy placentas. Among CONCEPTT participants, the slopes relating PlGF levels to birthweight z scores differed according to maternal glycaemia at 34weeks of gestation (p = 0.003). With optimal maternal glycaemia (HbA1c < 48mmol/mol [6.5%]/ or continuous glucose monitoring time above range ≤ 30%), birthweight z scores were reduced towards zero (normal weight) with increasing PlGF values (representing a healthy placenta), and increased with decreasing PlGF values. With suboptimal glycaemic status (HbA1c ≥ 48mmol/mol [6.5%] or time above range > 30%), increasing PlGF values were associated with heavier infants. Those with a healthy placenta (PlGF > 100) and suboptimal glycaemic control had a higher mean z score (2.45) than those with an unhealthy placenta (mean z score = 1.86). Similar relationships were seen when using sFlt-1/PlGF ratio as a marker for a healthy vs unhealthy placenta. In women with type 1 diabetes, infant birthweight is influenced by both glycaemic status and placental function. In women with suboptimal glycaemia, infant birthweight was heavier when placentas were healthy. Suboptimal placental function should be considered in the setting of suboptimal glycaemia and apparently 'normal' birthweight.

Soluble Flt-1, Placental Growth Factor, and Vascular Endothelial Growth Factor Serum Levels to Differentiate Between Active Lupus Nephritis During Pregnancy and Preeclampsia.

To evaluate mean serum levels of vascular endothelial growth factor (VEGF), placental growth factor (PlGF), and soluble Flt-1 (sFlt-1) in pregnant patients with systemic lupus erythematosus (SLE) with inactive disease, active lupus nephritis, and preeclampsia for differential diagnosis between these conditions. Pregnant women with SLE, with singleton pregnancies and no other autoimmune diseases, were classified according to disease activity (inactive SLE and active lupus nephritis) and the presence of preeclampsia. Serum samples were collected within 3 weeks of delivery and frozen for subsequent blinded analysis through the enzyme-linked immunosorbent assay method. A total of 71 women were included, with 41 classified as having inactive SLE (group 1; Systemic Lupus Erythematosus Pregnancy Disease Activity Index [SLEPDAI] score <4), 15 with a diagnosis of active lupus nephritis (group 2, SLEPDAI score ≥4, including renal criteria), and 15 with a diagnosis of preeclampsia (group 3). Patients in group 3 had higher mean levels of sFlt-1 and lower mean levels of PlGF compared to groups 1 and 2, both findings with statistical significance. The sFlt-1:PlGF ratio was also significantly higher in patients with preeclampsia, while mean VEGF levels were higher in pregnant woman with active lupus nephritis compared to patients with preeclampsia or inactive SLE. Evaluation of serum VEGF, PlGF, and sFlt-1 levels can differentiate between preeclampsia, inactive SLE, and active lupus nephritis during pregnancy.

MODULATION OF ANGIOGENIC BALANCE DURING PREGNANCY BY EXOGENOUS HEPARIN

Objective: Preeclampsia is a pregnancy-specific hypertensive disorder, commonly believed to result from placental dysfunction. Increased soluble vascular endothelial growth factor receptor-1 (sFlt-1) is consistently observed, and is believed to be pathogenic. sFlt-1 contains a binding site for the extracellular matrix (ECM) component heparan sulfate and, therefore, the anticoagulant heparin. Here we hypothesized that infusion of unfractionated heparin during pregnancy in rodents would displace and solubilize ECM-bound sFlt-1, cause maternal angiogenic imbalance, and maternal hypertension. Design and method: To determine the effects of heparin on sFlt-1 release in vitro. BeWo cytotrophoblasts and rodent placental explants were cultured at normoxic (8%) or hypoxic (1%) oxygen with or without exogenous heparin (10U/ml). Media sFlt-1 was measured by commercial ELISA, membrane-bound sFlt-1 was determined by quantitative immunofluorescence, and mRNA levels of Flt-1, sFlt-1, and heparanase were monitored by qRT-PCR. Pregnant rats were treated from GD14–19 with i.p. unfractionated heparin (300U/kg/day) or saline. MAP was monitored by carotid catheter on GD19, when tissues were collected and morphometrics noted. Placental and plasma sFlt-1 and VEGF were determined by commercial ELISA. Results: In BeWo cells, heparin increased media sFlt-1 in both normoxic (p = 0.0002) and hypoxic samples (p = 0.0066) compared to controls. Total Flt-1 and sFlt-1 was significantly decreased in heparin treated cells compared to controls (p = 0.0063) by immunofluorescence. In placental explants, 10U/mL heparin for 30 minutes significantly increased media sFlt-1 levels (p = 0.0005). Flt-1 mRNA was significantly decreased (p = 0.003) in heparin treated samples, while sFlt-1 and HPSE showed trends towards reduction. Infusion of unfractionated heparin increased MAP in pregnant rats compared to controls (110 ± 2.6 mmHg vs. 99.6 ± 1.3 mmHg, respectively; p = 0.0038). Placental mass (p = 0.0116) and pup weight (p = 0.0075) decreased with heparin treatment. Placental sFlt-1 levels were reduced with heparin (761.8 ± 48.4 pg/mg vs. 913.8 ± 33 pg/mg; p = 0.0178), while placental VEGF levels trended toward reduction. Heparin treatment reduced plasma bioavailable VEGF levels compared to controls (1174 ± 38.5 pg/mL vs. 1492 ± 62.2 pg/mL, respectively; p = 0.0006). Conclusions: Heparin infusion released ECM-bound sFlt-1, altered the angiogenic balance in the placenta and circulation of pregnant rats, and elevated their mean arterial pressure.

Effect of Fas/FasL signaling pathway activation in trophoblasts on recurrent spontaneous abortion.

To investigate the expression of Fas/FasL in human villous trophoblast cell HTR8-S/Vneo of patients with recurrent spontaneous abortion (RSA), and to explore the related function and molecular mechanism of Fas/FasL signaling pathway. The expression levels of FasL, Fas, and E-cadherin in the villous tissues of patients with RSA and those with artificial abortion in normal pregnancy (Normal) were detected by Western blot. CCK-8, flow cytometry, and wound healing were used to detect cell proliferation, apoptosis, and reactive oxygen species (ROS) level, and cell migration ability. Quantitative reverse transcription PCR (RT-qPCR) and Western blot were used to detect the expression of mRNA and protein of Notch1, FasL, Fas, E-cadherin, PKC, Hesl, sFlt-1, VEGF. Compared with normal group, the protein expression of FasL, Fas, and E-cadherin in villous tissues of RSA group were increased. HTR-8/SVneo cells in the H/R group had decreased proliferation and migration, increased apoptosis, and up-regulated ROS level compared with the Control group. The activation of Fas/FasL signaling pathway promoted HTR-8/SVneo cell injury in H/R group compared with the Fas/FasL+H/R group. Further RT-qPCR and Western blot experiments revealed that the mRNA and protein expression of Notch1, PKC, and Hesl were decreased in H/R group compared with Control group, while the mRNA and protein expression levels of E-cadherin, sFlt-1, and VEGF were significantly increased. The activation of Fas/FasL signaling pathway promotes trophoblast apoptosis induced by oxidative stress. This molecular mechanism relates to the inhibition of Notch1 signaling pathway activation, and the up-regulation of E-cadherin, sFlt-1, and VEGF expression.

Maternal Serum Inhibin-A Augments the Value of Maternal Serum PlGF and of sFlt-1/PlGF Ratio in the Prediction of Preeclampsia and/or FGR Near Delivery—A Secondary Analysis

Objective: We previously provided evidence to confirm that maternal serum levels of soluble Fms-like tyrosine kinase-1 (sFlt-1), placental growth factor (PlGF), and their ratio are useful tools to direct the management of preeclampsia (PE), fetal growth restriction (FGR), and PE+FGR near delivery. In this secondary analysis, we further examine the potential additive value of maternal serum Inhibin-A, which is a hormone marker of the transforming growth factor family, to the accuracy provided by maternal serum PlGF and sFlt-1. Methods: We conducted a secondary analysis where we extracted the data of a cohort of 125 pregnant women enrolled near delivery at the clinics of the University Medical Center of Ljubljana, Slovenia. The dataset included 31 cases of PE, 16 of FGR, 42 of PE+FGR, 15 preterm delivery (PTD), and 21 unaffected controls with delivery of a healthy baby at term. Cases delivered before 34 weeks’ gestation included 10 of PE, 12 of FGR, 28 of PE+FGR, and 6 of PTD. In addition to the recorded demographic characteristics and medical history and the maternal serum levels of PlGF and sFlt-1/PlGF ratio, which were previously published, we evaluated the added value of maternal serum Inhibin-A. The predictive accuracy of each biomarker, their ratios, and combinations were estimated from areas under the curve (AUC) of receiver operating characteristics (ROC) curves, Box and Whisker plots, and by multiple regression. We estimated accuracy by the continuous marker model and a cutoff model. Results: In this study, we combined Inhibin-A with PlGF or with the sFlt-1/PlGF ratio and showed a 10–20% increase in AUCs and 15–45% increase in the detection rate, at 10% false positive rate, of PE, and a lower, but significant, increase for PE+FGR and FGR in all cases but not for FGR in early cases delivered &lt; 34 weeks. The use of a cutoff model was adequate, although a bit higher accuracy was obtained from the continuous model. The highest correlation was found for PlGF with all three complications. Conclusion: In this secondary analysis, we have found that maternal serum Inhibin-A improves the accuracy of predicting PE and PE+FGR provided by maternal serum angiogenic markers alone, bringing the results to a diagnostic level; thus, it could be considered for directing clinical management. Inhibin-A had smaller or no added value for the accuracy of predicting FGR alone, mainly of early cases delivered &lt;34 weeks.

Evolution of Angiogenic Factors in Pregnant Patients with Breast Cancer Treated with Chemotherapy

Simple SummaryAnthracyclines and taxanes are being used as a standard treatment for breast cancer diagnosed during pregnancy. These chemotherapy regimens allow the continuation of pregnancy without delaying cancer treatment with relatively good maternal and neonatal outcomes. However, their effects on placental function and fetal development are not completely understood. Maternal serum angiogenic factors are a surrogate of placental function and are abnormal weeks before placental complications such as preeclampsia or intrauterine growth restriction development. In our cohort, pregnant women with breast cancer treated with chemotherapy during pregnancy show an antiangiogenic state with significantly higher levels of soluble fms-like tyrosine kinase (sFlt-1), sFlt-1/PGF ratio, and soluble endoglin (sEng) at the end of the third trimester. Angiogenic factors could be useful in the clinical obstetric management of these patients, although more studies are guaranteed.High prevalence of placental-derived complications, such as preeclampsia and intrauterine growth restriction, has been reported in women with breast cancer (BC) treated with chemotherapy during pregnancy (PBC-CHT). Aim: To ascertain whether PBC-CHT is associated with an imbalance of angiogenic factors, surrogate markers for placental insufficiency, that could explain perinatal outcomes. Methods: Prospective study between 2012 and 2016 in a single institution. Soluble fms-like tyrosine kinase (sFlt-1), placental growth factor (PlGF), and soluble endoglin (sEng) in maternal blood were assessed throughout pregnancy in 12 women with BC and 215 controls. Results: Cancer patients were treated with doxorubicin-based regimes and with taxanes. Ten PBC-CHT (83%) developed obstetrical complications. At the end of the third trimester, significantly higher levels of sFlt-1; sFlt-1/PGF ratio, and sEng levels were observed in BC women as compared to controls. Moreover; there was a significant correlation between plasma levels of sFlt-1 and the number of chemotherapy cycles administered. Besides, more chemotherapy cycles correlated with lower birthweight and head circumference at birth. Conclusions: Women with BC treated during pregnancy showed an antiangiogenic state compatible with placental insufficiency. Angiogenic factors could be useful in the clinical obstetric management of these patients; although further studies will be required to guide clinical decision-making.

SERUM LEVELS OF SOLUBLE FMS –LIKE TYROSINE KINASE -1 IN PRE-ECLAMPTIC AND ECLAMPTIC CASES AND ITS CORRELATION TO FETOMATERNAL OUTCOME

Preeclampsia is a multisystem progressive disorder characterized by the new onset of hypertension and proteinuria or the new onset of hypertension and significant end-organ dysfunction with or without proteinuria after 20 weeks of gestation or postpartum in previously normotensive women. It is develops due to placental and maternal vascular dysfunction and disappears after delivery gradually by time. The incidence is estimated to be between 3 and 10% of all pregnancies.Maternal endothelial dysfunction which leads to pre-eclamptic signs and symptoms is induced by high levels of the antiangiogenic factor sFLT1, which is produced in the placenta and released into the maternal circulation. sFLT1 is a soluble splice variant of the membrane-bound receptor VEGFR1 that binds to the proangiogenic proteins VEGF and placental growth factor (PlGF); therefore, sFLT1 acts as a ligand trap and antagonizes ligand-mediated angiogenic signalling via the cell surface receptors. Aim of the workThe aim of this work was to evaluate serum levels of soluble fms-like tyrosine kinase-1 (sFlt-1) in cases of severe preeclampsia and eclamptic fits,and to correlate serum levels of soluble fms-like tyrosine kinase-1 (sFlt-1) in these cases to outcome of fetus and mother and incidence of complication.

Soluble syndecan-1 and glycosaminoglycans in preeclamptic and normotensive pregnancies

Preeclampsia, an important cause of maternal and fetal morbidity and mortality, is associated with increased sFLT1 levels and with structural and functional damage to the glycocalyx contributing to endothelial dysfunction. We investigated glycocalyx components in relation to preeclampsia in human samples. While soluble syndecan-1 and heparan sulphate were similar in plasma of preeclamptic and normotensive pregnant women, dermatan sulphate was increased and keratan sulphate decreased in preeclamptic women. Dermatan sulphate was correlated with soluble syndecan-1, and inversely correlated with blood pressure and activated partial thromboplastin time. To determine if syndecan-1 was a prerequisite for the sFlt1 induced increase in blood pressure in mice we studied the effect of sFlt1 on blood pressure and vascular contractile responses in syndecan-1 deficient and wild type male mice. The classical sFlt1 induced rise in blood pressure was absent in syndecan-1 deficient mice indicating that syndecan-1 is a prerequisite for sFlt1 induced increase in blood pressure central to preeclampsia. The results show that an interplay between syndecan-1 and dermatan sulphate contributes to sFlt1 induced blood pressure elevation in pre-eclampsia.

Predictive values of various serum biomarkers in women with suspected preeclampsia: A prospective study.

BackgroundPreeclampsia (PE) prediction has been shown to improve the maternal and fetal outcomes in pregnancy. We aimed to evaluate the PE prediction values of a series of serum biomarkers.MethodsThe singleton pregnant women (20–36 gestational weeks) with PE‐related clinical and/or laboratory presentations were recruited and had the blood drawn at their first visits. The following markers were tested with the collected serum samples: soluble fms‐like tyrosine kinase 1 (sFlt‐1), placental growth factor (PlGF), thrombomodulin (TM), tissue plasminogen activator inhibitor complex (tPAI‐C), complement factors C1q, B, H, glycosylated fibronectin (GlyFn), pregnancy‐associated plasma protein‐A2 (PAPP‐A2), blood urea nitrogen (BUN), creatinine (Cre), uric acid (UA), and cystatin C (Cysc).ResultsOf the 196 recruited subjects, 25% (n = 49) developed preeclampsia before delivery, and 75% remained preeclampsia negative (n = 147). The serum levels of sFlt‐1, BUN, Cre, UA, Cysc, and PAPP‐A2 were significantly elevated, and the PlGF level was significantly decreased in the preeclampsia‐positive patients. In the receiver operating characteristics (ROC) analyses, the area under the curves were listed in the order of decreasing values: 0.73 (UA), 0.67 (sFlt‐1/PlGF), 0.66 (Cysc), 0.65 (GlyFn/PlGF), 0.64 (PAPP‐A2/PlGF), 0.63 (BUN), 0.63 (Cre), and 0.60 (PAPP‐A2). The positive predictive values of these serum markers were between 33.1% and 58.5%, and the negative predictive values were between 80.9% and 89.5%.ConclusionsThe serum markers investigated in current study showed better performance in ruling out than ruling in PE. Absence of pre‐defined latency period between blood draw and the onset of PE limits the clinical utility of these markers.

Crocin exhibits an antihypertensive effect in a rat model of gestational hypertension and activates the Nrf-2/HO-1 signaling pathway.

Gestational hypertension is a leading cause of both prenatal and maternal mortality and morbidity; however, there have been rather limited advances in the management of gestational hypertension in recent years. There has been evidence supporting the antihypertensive properties of crocin, but the specific mechanism is still unclear. N-Nitro-L-arginine methyl ester (L-NAME) was employed to establish a rat model with a preeclampsia-like phenotype, particularly gestational hypertension. Enzyme-linked immunosorbent assays were conducted to determine the levels of placental growth factor (PlGF) and soluble fms-like tyrosine kinase (sFlt-1); the levels of the circulating cytokines interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α; and oxidative stress factors. Quantitative RT-PCR assays were performed to assess the transcript levels of various cytokines in the placenta, and western blot assays were carried out to evaluate the protein levels of heme oxygenase-1 (HO-1) and nuclear factor-erythroid 2-like 2 (Nrf-2). Treatment with crocin reduced the blood pressure of rats with gestational hypertension, which was accompanied by suppressed circulating levels of PlGF and sFlt-1. Crocin further alleviated the inflammatory signals and oxidative stress in the serum, as well as in placental tissues, in rats with L-NAME-induced hypertension. Crocin treatment also improved pregnancy outcomes in terms of fetal survival, fetal weight, and the fetal/placental weight ratio. Finally, in hypertension elicited by L-NAME, crocin stimulated the placental Nrf-2/HO-1 pathway. Crocin alleviated inflammatory and oxidative stress in placental tissues, thereby protecting against gestational hypertension, one of the major phenotypes of preeclampsia, and activated the Nrf-2/HO-1 pathway.

Biomarkers and Their Relation to Cardiac Function Late After Peripartum Cardiomyopathy

BackgroundAngiogenic imbalance involving the placental protein soluble Fms-like tyrosine kinase-1 (sFlt-1) and cleavage of the nursing-hormone prolactin by the enzyme cathepsin D (CD) both play a role in the pathogenesis of peripartum cardiomyopathy (PPCM). We hypothesized that angiogenic imbalance and increased activity of CD have a long-lasting impact in women with PPCM. Methods and ResultsA nationwide Danish cohort of women with PPCM (PPCM group, n = 28), age matched women with previous preeclampsia (n = 28) and uncomplicated pregnancies (n = 28) participated in a follow-up study including biomarker analysis, exercise testing and cardiac magnetic resonance imaging. The median time to follow-up was 91 months (range 27–137 months) for the PPCM group. Levels of sFlt-1, placental growth factor, N-terminal pro-natriuretic brain peptide, and copeptin were all significantly higher in the PPCM group. More women in the PPCM group had detectable CD activity (68%) compared with the preeclampsia group (29%) and uncomplicated pregnancies group (36%) (P = .0002). Levels of angiogenic factors and biomarkers correlated inversely with maximal exercise capacity and cardiac functional parameters assessed with cardiac magnetic resonance imaging. ConclusionsWomen with PPCM had higher biomarker levels and CD activity up to 7 years after diagnosis. Higher biomarker levels correlated inversely with maximal exercise capacity and markers of cardiac dysfunction suggesting that persistent angiogenic imbalance and increased CD activity is associated with residual cardiac dysfunction.

Circulating growth factors and cardiac remodeling in the community: The Framingham Heart Study

Background and aimsCardiac and vascular growth factors (GF) may influence myocardial remodeling through cardiac growth and angiogenic effects. We hypothesized that concentrations of circulating GF are associated with cardiac remodeling traits. MethodsWe related blood concentrations of vascular endothelial GF (VEGF), VEGFR-1 (sFlt1), angiopoietin 2 (Ang-2), soluble angiopoietin type-2 receptor (sTie2), hepatocyte GF (HGF), insulin-like GF (IGF)-1, IGF binding protein (IGFBP)-3, and growth differentiation factor-15 (GDF-15) to echocardiographic traits in 3151 Framingham Study participants (mean age 40 years, 55% women). We evaluated the following measures: left ventricular (LV) mass index (LVMi), LV ejection fraction (LVEF), global longitudinal strain (GLS), mitral E/e', and aortic root diameter (AoR). All biomarker values were sex-standardized. ResultsIn multivariable-adjusted analyses, higher GDF-15 concentrations were associated with higher log-LVMi (β = 0.009 per SD, P = 0.01). Similarly, sTie2 concentrations were positively associated with log-E/e' (β = 0.011 per SD, P = 0.04). IGF-1 and Ang-2 concentrations were positively and negatively associated with GLS, respectively (βIGF-1 = 0.16 per SD and βAng-2 = −0.15 per SD, both P < 0.05), whereas higher sFlt1 and Ang-2 levels were associated with smaller log-AoR (βsFlt1 = −0.004 per SD and β Ang-2 = −0.005 per SD, respectively; P < 0.05). ConclusionIn our large community-based sample, we observed patterns of associations between several circulating vascular GF and cardiac remodeling indices that are consistent with the known biological effects of these pro- and anti-angiogenic factors on the myocardium and conduit arteries. Additional studies are warranted to replicate our findings and assess their prognostic significance.

Pravastatin Promotes Endothelial Colony-Forming Cell Function, Angiogenic Signaling and Protein Expression In Vitro.

Endothelial dysfunction is a primary feature of several cardiovascular diseases. Endothelial colony-forming cells (ECFCs) represent a highly proliferative subtype of endothelial progenitor cells (EPCs), which are involved in neovascularization and vascular repair. Statins are known to improve the outcome of cardiovascular diseases via pleiotropic effects. We hypothesized that treatment with the 3-hydroxy-3-methyl-glutaryl–coenzyme A (HMG-CoA) reductase inhibitor pravastatin increases ECFCs’ functional capacities and regulates the expression of proteins which modulate endothelial health in a favourable manner. Umbilical cord blood derived ECFCs were incubated with different concentrations of pravastatin with or without mevalonate, a key intermediate in cholesterol synthesis. Functional capacities such as migration, proliferation and tube formation were addressed in corresponding in vitro assays. mRNA and protein levels or phosphorylation of protein kinase B (AKT), endothelial nitric oxide synthase (eNOS), heme oxygenase-1 (HO-1), vascular endothelial growth factor A (VEGF-A), placental growth factor (PlGF), soluble fms-like tyrosine kinase-1 (sFlt-1) and endoglin (Eng) were analyzed by real time PCR or immunoblot, respectively. Proliferation, migration and tube formation of ECFCs were enhanced after pravastatin treatment, and AKT- and eNOS-phosphorylation were augmented. Further, expression levels of HO-1, VEGF-A and PlGF were increased, whereas expression levels of sFlt-1 and Eng were decreased. Pravastatin induced effects were reversible by the addition of mevalonate. Pravastatin induces beneficial effects on ECFC function, angiogenic signaling and protein expression. These effects may contribute to understand the pleiotropic function of statins as well as to provide a promising option to improve ECFCs’ condition in cell therapy in order to ameliorate endothelial dysfunction.

Placenta Growth Factor and Soluble Fms-Like Tyrosine Kinase 1 in Preeclampsia and Normotensive Pregnant Nigerian Women

Background: Preeclampsia (PE) is still one of the leading causes of maternal/perinatal morbidity/mortality in Nigeria. Imbalance between placenta growth factor (PLGF) and soluble fms-like tyrosine kinase 1 (sFlt1) has been reportedly present both before and after the manifestation of PE; however, Nigerian data regarding these angiogenesis-related substances are lacking. We here attempted to determine the maternal serum level of PLGF and sFlt1 and sFlt1/PLGF ratio in PE vs. non-PE women in Lagos State University Teaching Hospital, Nigeria. Methods: An observational cross-sectional study was made on 75 women with PE and 75 age-gestational-age matched women without PE, as case and control, respectively. Levels of sFlt-1, PIGF and the sFlt-1: PIGF ratio was compared between the two. Results: Serum levels of Flt-1 and sFlt1/PIGF ratio were significantly higher in PE patients (6581.86 ± 865.75, and 146.42 ± 92.43) than in the normotensive control (4584.52 ± 1479.6 and 11.60 ± 6.42). PIGF was significantly lower in PE patients (70.14 ± 51.03) than the normotensives (494.06 ± 475.8). There were positive and negative correlations between the sFlt-1 and PLGF respectively and mean arterial blood pressure. Conclusion: Serum sFlt-1, sFlt1/PIGF ratio was significantly higher and PIGF levels were significantly lower in PE than normotensive control in Nigerian population.

Clinical value of serum VEGF and sFlt-1 in pernicious placenta previa

This study was designed to explore the expression and the diagnostic value of vascular endothelial growth factor (VEGF) and soluble fms-like tyrosine kinase-1 (sFlt-1) in pernicious placenta previa (PPP) combined placental accreta/increta. A total of 140 PPP patients were enrolled and divided into two groups: 56 patients with placenta accreta/increta (PA group), and 84 patients without placenta accreta/increta (non-PA group). In the same period, 46 pregnant women without PPP who had undergone caesarean section were selected as controls. The levels of VEGF and sFlt-1 in serum were detected by enzyme-linked immunosorbent assay. Diagnostic efficiency of VEGF and sFlt-1 in serum were evaluated by receiver operating characteristics curve. It was found that both VEGF and sFlt-1 had diagnostic value for PPP and placenta accreta/increta combined PPP. In addition, the levels of VEGF and sFlt-1 could be used to distinguish placenta accreta from placenta increta. VEGF was negatively correlated with sFlt-1 in PPP patients. In summary, the levels of VEGF and sFlt-1 could be used as auxiliary indicators to diagnose PPP and distinguish between placenta accreta and increta. KEY POINTS The levels of VEGF and sFlt-1 could be used to distinguish placenta accreta from placenta increta. VEGF is negatively correlated with sFlt-1 in PPP patients. The levels of VEGF and sFlt-1 could be used as auxiliary indicators to diagnose PPP and distinguish between placenta accreta and increta.

Antiangiogenic Biomarker Soluble Fms-like Tyrosine Kinase-1 in Pregnancy Complicated with Preeclampsia: A Cohort Study

Introduction: Preeclampsia (PE) is one of the leading obstetric diseases with significant morbidity and mortality in both mother and foetus. The etiology of preeclampsia is unknown. It may result from several reasons with imbalance between angiogenic regulatory factors in maternal circulation as one of the factor. High circulatory levels of Soluble Fms-like Tyrosine Kinase-1 (sFlt-1) are detectable several weeks before clinical presentation of preeclampsia. Aim: To determine association of serum levels of sFlt-1 with preeclampsia in second and third trimester of pregnancy. Materials and Methods: A prospective cohort study was conducted in the Department of Biochemistry in collaboration with the Department of Obstetrics and Gynaecology in a tertiary care hospital (Sri Guru Ram Das Institute of Medical Sciences and Research, Amritsar, Punjab, India), from November 2018 to March 2021. The study participants were divided into two groups i.e, normotensive (group 1) and preeclamptic (group 2). The enrollment of participants was done during second trimester. Serum sFlt-1 concentration was measured in second trimester (24-28 weeks) and in third trimester (beyond 28 weeks) using Enzyme Linked Immunoassay (ELISA) kits. Receiver Operating Characteristics (ROC) curve analysis was done for evaluation of Area Under Curve (AUC), sensitivity and specificity using software defined cut-off values. Results: Total 60 participants were there in each group with maternal mean age of 25.8±3.2 years in group 1 and 30.6±5.5 years in group 2 (p-value &lt;0.001). sFlt-1 levels were significantly higher in preeclampsia group both in second and third trimesters when compared with normotensive group with median 313.07 versus 65.150 (p-value &lt;0.001) and 337.875 versus 76.925 (p-value &lt;0.001), respectively. The ROC curve analysis using 190.5 ng/mL as cut-off point in second trimester showed sensitivity 90%, specificity 85%, AUC was 0.832, 95% CI (0.745-0.918) and in third trimester at cut-off point 271.5 ng/mL showed sensitivity 90%, specificity 90%, AUC was 0.884, 95% CI (0.817-0.951). Conclusion: The soluble Fms-Like Tyrosine Kinase-1-1 (sFlt-1), may serve as biomarker for early diagnosis and can improve prediction of preeclampsia.

Increased Expression Levels of Soluble Fms-Like Tyrosine Kinase-1 Inhibits Nervous System Development of Zebrafish

Introduction Preeclampsia (PE) remains a leading cause of maternal and perinatal morbidity. At present, the treatments of preeclampsia are limited, and many patients need to terminate the pregnancy to relieve the disease. Soluble fms-like tyrosine kinase-1 (sFlt-1) is a decoy receptor of placental growth factor (PIGF) and vascular endothelial growth factor which can promote angiogenesis. Its level increases throughout pregnancy with preeclampsia and umbilical cord blood of their newborns. Material and methods In this experiment, we constructed zebrafish with high expression levels of sFlt-1 and explored their changes of nervous system developments and behaviors. Results Comparing with the WT group and the casRx group, the cerebral vascular developments at 72 h in the F1、F2 group were significantly decreased, the ratios of brain volume area to whole body area at 96 h were significantly decreased, and the behavioral trajectory distances and movement speeds at 6th day were shortened and slowed down. Conclusions The increased expression levels of sFlt-1 in zebrafish can inhibit the developments of cerebral blood vessels, brain volumes and the behavioral activities. It is speculated that elevated sFlt-1 in the pathological state of PE can inhibit the developments of nervous system in offspring.