Production Of Antiangiogenic Factors Research Articles

A Novel Dual-Function Redox Modulator Relieves Oxidative Stress and Anti-Angiogenic Response in Placental Villus Explant Exposed to Hypoxia-Relevance for Preeclampsia Therapy.

Preeclampsia (PE) is a severe, life-threatening complication during pregnancy (~5-7%), and no causative treatment is available. Early aberrant spiral artery remodeling is associated with placental stress and the release of oxygen radicals and other reactive oxygen species (ROS) in the placenta. This precedes the production of anti-angiogenic factors, which ultimately leads to endothelial and trophoblast damage and the key features of PE. We tested whether a novel dual-function redox modulator-AKT-1005-can effectively reduce placental oxidative stress and alleviate PE symptoms in vitro. Isolated human villous explants were exposed to hypoxia and assessed to determine whether improving cell-redox function with AKT-1005 diminished ROS production, mitochondrial stress, production of the transcription factor HIF1A, and downstream anti-angiogenic responses (i.e., sFLT1, sEng production). MitoTEMPO was used as a reference antioxidant. In our villous explant assays, pretreatment with AKT-1005 reduced mitochondrial-derived ROS production, reduced HIF-1A, sFLT1, and sEng protein expression, while increasing VEGF in hypoxia-exposed villous trophoblast cells, with better efficiency than MitoTEMPO. In addition, AKT-1005 improved mitochondrial electron chain enzyme activity in the stressed explant culture. The redox modulator AKT-1005 has the potential to intervene with oxidative stress and can be efficacious for PE therapy. Future studies are underway to assess the in vivo efficacy of HMP.

Abstract P105: Redox Modulator Akt-1005 Alleviates Oxidant Stress In An In Vitro Model Of Preeclampsia

Background: Preeclampsia (PE) is a pregnancy-specific syndrome affecting 5-7% of all pregnant women, and there is no effective treatment available. Early abnormal placental development is associated with oxidative stress and the release of reactive oxygen species in the placenta. This phenomenon leads to downstream signaling and production of anti-angiogenic factors, which cause endothelial and trophoblast dysfunction and the cardinal features of PE, i.e., hypertension, proteinuria, and in severe cases, eclampsia. Our group developed a novel series of redox modulators and sought to test if these compounds can effectively reduce placental oxidative stress and mitigate PE symptoms in vitro . Methods: We induced in vitro oxidant stress in human villous explants with 2% O 2 and assessed whether augmenting cell-redox function with the proposed compound AKT-1005 reduced (i) cell injury (cell cytotoxicity assay), (ii) mitochondrial stress (mitochondrial -derived superoxide production, mitochondria dysfunction) (iii) production of the transcription factor HIF1A and (iv) downstream anti-angiogenic responses (sFLT1 production). These effects were compared with reference antioxidants i.e., Vit C and MitoTEMPO. Results: In our villous explant assays, pretreatment with the AKT-1005 reduced mitochondrial-derived ROS production in hypoxia-exposed trophoblasts cells, indicating that the key factor in the development of PE, the oxidant stress, can be alleviated by the redox modulator compound. The AKT-1005 pretreatment also reduced HIF-1A expression and sFLT1 protein expression in hypoxia-exposed villous trophoblast cell, with better efficacy than Vit C and MitoTempo. Furthermore, AKT-1005 improved the mitochondrial electron chain enzyme activity in the stressed explant culture, mainly in syncytiotrophoblast cells, which is another promising characteristic of the applied antioxidant molecule. Conclusion: In reducing placental trophoblast oxidative stress, redox modulators present a potential novel therapeutic approach for the treatment of preeclampsia. Future investigation is warranted regarding the in vivo use of these compounds.

SIRT 1 Activator Loaded Inhaled Antiangiogenic Liposomal Formulation Development for Pulmonary Hypertension.

Pulmonary hypertension (PH) is characterized by the rise in mean pulmonary arterial pressure (≥ 20mmHg at rest) due to the narrowing of the pulmonary arterial networks. Current treatments provide symptomatic treatment and the underlying progress of PH continues leading to higher mortality rates due to non-reversal of the disease. This warrants the need for drug therapies targeting angiogenesis and vascular remodeling mechanisms. Resveratrol, SIRT 1 activator, alters various signaling pathways, inhibits apoptosis, and negatively regulates angiogenesis either by increasing the production of anti-angiogenic factors or inhibiting pro-angiogenic factors. Our work describes the liposomal formulation development, physicochemical characterization, and in vitro aerosolization performance of resveratrol liposomal dry powder formulation. The resveratrol liposomal dry powder formulation reduces the right ventricular systolic pressure measured during right jugular vein catheterization and significantly reverses the PH disease pathological changes as demonstrated by histological observations of pulmonary arterial lumen and ventricular hypertrophy. The developed resveratrol liposomal dry powder formulation alleviates the pulmonary arterial remodeling through its antiangiogenic mechanism and indicates a promising therapeutic strategy for PH treatment.

Does COVID-19 cause pre-eclampsia?

Does COVID-19 cause pre-eclampsia?

Uncaria tomentosa extract (AC-11) improves pregnancy hypertension together with suppression of sFlt-1 and sEng.

Disruption of well-controlled reproductive functions leads to pregnancy complications such as hypertensive disorders of pregnancy (HDP). Uncaria tomentosa (Wild), known as cat's claw, is widely used for the treatment of a various types of health problems; AC-11 (AC-11®, hot-water extract of U. tomentosa) is unique phytochemical compound and has potential roles as anti-inflammatory or anti-oxidant processes. We investigated whether AC-11 has a protective effect on pathogenesis of HDP in vivo and production of anti-angiogenic factors (sFlt-1 and sEng, major factors for the onset of HDP) in in vitro. Non-pregnant or pregnant mice were administered AC-11 (4mg/mL), then, angiotensin II (Ang II) was subcutaneously infused to increase blood pressure. Human placental tissues or human umbilical vein endothelial cells (HUVECs) were incubated with or without AC-11. Treatment with AC-11 significantly reduced blood pressure induced by Ang II infusion. The population of CD8+T cells, the ratio of CD8/CD4, and plasma interleukin-6 levels were increased by Ang II infusion, and were decreased by AC-11 both in pregnant and non-pregnant mice. In pregnant mice, plasma levels of sFlt-1 and sEng were decreased by AC-11. In in vitro cell culture of HUVECs or placental tissue culture, treatment with AC-11 significantly inhibited secretion of sFlt-1 and sEng. We suggest a novel role of AC-11 in regulating blood pressure by controlling the balance of T cell population and inflammatory cytokine production both in non-pregnant and pregnant conditions. In addition, AC-11 inhibits HDP-related factors, including sFlt-1 and sEng, suggesting that AC-11 may useful for relieving HDP.

Abstract P236: Organofluorine Hydrazones Preventing Oxidant Stress In An In Vitro Model Of Preeclampsia

Background: Preeclampsia (PE) is a pregnancy-specific syndrome affecting 5-7% of all pregnant women, and there is no effective treatment available. Early abnormal placental development is associated with oxidative stress and the release of reactive oxygen species in the placenta. This phenomenon leads to downstream signaling and production of anti-angiogenic factors, which cause endothelial and trophoblast dysfunction and the cardinal features of PE, i.e., hypertension, proteinuria, and in severe cases, eclampsia. Our group developed a novel series of mitochondria-targeted antioxidants and sought to test if these compounds can effectively reduce placental oxidative stress and mitigate PE symptoms in vitro. Methods: We induced in vitro oxidant stress in human trophoblast (HTR8/SVneo) cells with hydrogen peroxide (H 2 O 2 ) and assessed whether augmenting cell-redox function with the proposed antioxidant compounds reduced (i) cell injury (cell cytotoxicity assay), (ii) mitochondrial stress (mitochondrial -derived superoxide production, mitochondria dysfunction) (iii) production of the transcription factor HIF1A and (iv) downstream anti-angiogenic responses (sFLT1 production). These effects were compared with the antioxidants - NAC, Vit E and MitoQ. Results: In our cell-based assays, pretreatment with the hydrazone compounds reduced mitochondrial-derived ROS production in H 2 O 2 -exposed trophoblasts cells, indicating that the key factor in the development of PE, the oxidant stress, can be alleviated by the antioxidant hydrazones. The most effective of these compounds, HY12 also reduced HIF-1A expression and sFLT1 protein expression H 2 O 2 -exposed HTR8 cells. Furthermore, the antioxidant hydrazones improved the mitochondrial electron chain enzyme activity in the stressed HTR8 cells, which is another promising characteristic of the applied hydrazones. Conclusion: In reducing placental trophoblast oxidative stress, hydrazone-based antioxidants present a potential novel therapeutic approach for the treatment of preeclampsia. Future investigation is warranted regarding the in vivo use of these compounds.

STING agonist-based treatment promotes vascular normalization and tertiary lymphoid structure formation in the therapeutic melanoma microenvironment

BackgroundThe degree of immune infiltration in tumors, especially CD8+ T cells, greatly impacts patient disease course and response to interventional immunotherapy. Enhancement of tumor infiltrating lymphocyte (TIL) is a critical...

Sulforaphane improves vascular reactivity in mouse and human arteries after “preeclamptic-like” injury

IntroductionThe widespread maternal endothelial dysfunction that underlies the manifestations of preeclampsia is thought to arise from excessive placental production of antiangiogenic factors and enhanced oxidative stress. Therefore, we assessed whether the natural antioxidant sulforaphane could improve vascular function. MethodsCell viability of human umbilical vein endothelial cells (HUVECs) was assessed after 24 or 48 h in normoxia (20% O2) or hypoxia (1% O2) with or without sulforaphane. To model vascular dysfunction associated with preeclampsia, mouse mesenteric arteries were incubated in trophoblast conditioned media (TCM), and human omental arteries incubated in preeclamptic explant media (PEM) with or without sulforaphane. Both media are rich in antiangiogenic compounds associated with preeclampsia. TCM was generated from primary cytotrophoblast cells from term placentae of normotensive, while PEM was generated from explants from preeclamptic women. Reactivity was assessed by wire myography. sulforaphane's actions as a vasodilator were also investigated. ResultsUnder conditions of hypoxia, sulforaphane improved HUVEC viability. In mouse mesenteric arteries, sulforaphane reduced contraction evoked by potassium (p < 0.001), phenylephrine and endothelin 1 (all p < 0.001). Sulforaphane also inhibited Ca2+-induced contraction (p = 0.014). Sulforaphane prevented TCM-induced augmentation of phenylephrine and angiotensin II-mediated contraction of mouse mesenteric arteries. In human omental arteries, sulforaphane induced vasodilation (p < 0.001), and prevented PEM-induced endothelial dysfunction by restoring arterial sensitivity to the endothelium-dependent vasodilator bradykinin (p = 0.008). DiscussionSulforaphane causes relaxation in arteries and protects against arterial dysfunction induced by placental-derived antiangiogenic factors, which are known to contribute to the preeclampsia.

Epigallocatechin Gallate (EGCG) Improves Anti-Angiogenic State, Cell Viability, and Hypoxia-Induced Endothelial Dysfunction by Downregulating High Mobility Group Box 1 (HMGB1) in Preeclampsia.

BackgroundPreeclampsia (PE) is a serious complication of pregnancy with no effective therapy. This study assessed whether epigallocatechin gallate (EGCG) could reduce the production of anti-angiogenic factors, improve cell viability, and suppress endothelial dysfunction in vitro via regulating high mobility group box 1 (HMGB1) in preeclampsia.Material/MethodsHuman umbilical vein endothelial cells (HUVECs) grown in conditioned medium from hypoxic JEG-3 cells were used to investigate the effects of EGCG on anti-angiogenic state, cell viability, and markers of endothelial dysfunction. To confirm that EGCG exerted its effects via HMGB1, we also examined the impact of EGCG on anti-angiogenic state, cell viability, and endothelial dysfunction following HMGB1 treatment in vitro.ResultsEGCG inhibited HMGB1 expression in hypoxic trophoblast cells in a dose-dependent manner. In addition, EGCG relieved anti-angiogenic state and endothelial dysfunction in hypoxic trophoblast cells by downregulating HMGB1. Moreover, EGCG dose-dependently promoted cell proliferation by downregulating HMGB1.ConclusionsTaken together, our data show the protective role of EGCG in preeclampsia and revealed EGCG-mediated effects on the production of anti-angiogenic factors, cell viability, and endothelial dysfunction through downregulating HMGB1. These observations suggest that EGCG is a novel therapeutic candidate for preeclampsia.

Sulforaphane improves endothelial function and reduces placental oxidative stress in vitro.

The maternal endothelial dysfunction characteristic of preeclampsia arises, in part, from excessive placental production of anti-angiogenic factors, including soluble Flt-1, soluble endoglin and activin A, inducing oxidative stress. We assessed whether the antioxidant and NRF2-activator sulforaphane could mitigate endothelial and trophoblast dysfunction in vitro. We induced dysfunction in human umbilical vein endothelial cells (HUVECs) with TNF-α, assessing endothelial activation and dysfunction (endothelin-1, vascular cell adhesion molecule; VCAM1, intracellular adhesion molecule; ICAM1, e-selectin and endothelial permeability) in the presence or absence of sulforaphane. We also assessed the effects of sulforaphane in mitigating hypoxic and hyperoxic injury in term placental explants by measuring secretion of anti-angiogenic factors. To assess the role of NRF2 we silenced NRF2 in HUVECs and primary trophoblast cells. Sulforaphane reduced TNF-α mediated HUVEC secretion of endothelin-1, VCAM1, ICAM1 and E-selectin, and prevented increased endothelial permeability. In placental explants, sulforaphane reduced the secretion of soluble Flt-1, soluble endoglin and activin A. Sulforaphane induced activation and nuclear translocation of NRF2 in HUVECs, inducing heme oxygenase 1. NRF2 silencing blocked some but not all of sulforaphane's effects in HUVECs. NRF2 silencing did not prevent sulforaphane's inhibition of trophobast secretion of soluble Flt-1 or activin A. In reducing placental and endothelial oxidative stress, sulforaphane may offer a new adjuvant therapeutic approach for the treatment of preeclampsia.

Differences in levelFms-Like Tyrosine Kinase-1 (sFlt-1), soluble Endoglin (s-Eng), and Placental Growth Factor (PIGF) between Early Onset Preeclampsia and Late Onset Preeclampsia

Early Onset Preeclampsia (EO-PE) is preeclampsia that develops before 34 weeks 'gestation, caused by intrinsic factors, while Late Onset Preeclampsia (LO-PE) is preeclampsia that develops after 34 weeks' gestation due to extrinsic and maternal factors. There is an increased production of antiangiogenic factors (sFlt-1, s-Eng and PIGF) contribute to pathophysiology of preeclampsia.This study aims to measure the difference of sFlt-1, sEng, PIGF levels between EO-PE and LO-PE. This was an observational study with cross sectional design conducted at Dr. M. Djamil, TK Hospital. III dr. Reksodiwiryo and Biomedical Laboratory FK Unand Padang from August 2017 to August 2018. The sample of this study were 26 severe preeclampsia women : 13 (EO-PE) and 13 (LO-PE), selected using consecutive sampling. Levels of sFlt-1, sEng, PIGF were examined using the enzyme-linked immunosorbent assay (ELISA) method. Statistical analysis was performed using unpaired t test and Mann-Whitney Test. Results shown that serum levels of sFlt-1 and sEng in (EO-PE) were 9.51 ± 0.71 ng / L, 1.44 ± 0.06 ng / mL, 5.79 ± 0.42 ng / mL while in PEAL it was 8, 89 ± 0.78 ng / mL, 1.35 ± 0.14 ng / mL, 6.72 ± 0.76. There were a significant difference with a value of p &lt;0.05. The conclusion of this study is that the levels of sFlt-1 and sEng are higher in (EO-PE) than(LO-PE)and PIGF levels was lower in (EO-PE) compared to (LO-PE)

89. Placental oxidative stress, angiogenic factor and maternal endothelial function in pregnant women with normotensive fetal growth restriction

Introduction Placental oxidative damage may increase the production of anti-angiogenic factors, resulting in maternal endothelial dysfunction in pregnant women with preeclampsia (PE). Endothelial dysfunction is observed in women with a history of normotensive fetal growth restriction (FGR). However, placental oxidative stress, angiogenic factors and maternal endothelial function in those women remain unclear. Objectives This study aimed to assess the relationship between placental oxidative stress, angiogenic factors and maternal endothelial function in pregnant women with normotensive FGR. Methods A total of 21 women with uncomplicated pregnancies, 17 women with early-onset preeclampsia (PE), 18 with late-onset PE, and 14 with normotensive FGR were evaluated. We measured to determine serum parameters of oxygen free radicals (d-ROMs), PlGF and sFlt-1 as maternal anti-angiogenic factor, placental oxidative DNA damage, and flow-mediated vasodilation (FMD) as a marker of maternal endothelial function. Immunohistochemical analysis was performed to measure the proportion of placental trophoblast cell nuclei staining positive for 8-hydroxy-2′-deoxyguanosine (8-OHdG) and redox factor-1(ref-1), which are markers of oxidative DNA damage. Results Maternal serum d-ROMs, sFlt-1 concentrations, and FMD did not significantly differ between control and normotensive FGR groups. The proportion of nuclei staining positive for 8-OHdG was significantly higher in the normotensive FGR group relative to the control group, but ref-1 were not significantly differences among all groups. Conclusion Our findings demonstrate that although placental oxidative DNA damage was observed in both women with PE and those with normotensive FGR, pregnant women with normotensive FGR show no increase in the concentrations of sFlt-1 and d-ROMs, or a decrease in FMD.

Apium graveolens Prevents Intrauterine Growth Restriction via Suppression of Antiangiogenic Factor Production

Preeclampsia is the worldwide leading cause of fetomaternal morbidity and mortality which involves the placental dysfunction. A poor placentation and formed of non-dilated spiral artery caused utero-placental circulation insufficiency, resulted in inadequate supply of nutrients and oxygen to support normal aerobic growth of the fetus. Apium graveolens or celery has been widely known as antioxidant, antiinflammation and antihypertensive with flavonoid-apigenin as main active compound. Apigenin can inhibit TNF-α, HIF-1α and nitric oxide blocking as major pathophysiological pathway of preeclampsia. This study was aimed to find how the Apium graveolens can improve intrauterine growth and its correlation with suppression of anti-angiogenic factor sFlt-1 in anti-Qa2 preeclampsia animal model. Twenty female BALB/c Mus musculus were divided into 4 groups: control, anti-Qa2 and anti-Qa2 with 500 and 1000 mg/kgBW celery herbs extract. The fetal weights and lengths, placental weights and serum sFlt-1 levels were measured and analyzed with One Way ANOVA and further tested with Least Significance Difference in 95% confidence interval. The result showed a difference between control and treatments group (p≤0.05) with 1000 mg/kgBW significantly increase intrauterine growth and decrease sFlt-1, then there is a negative correlation between intrauterine weight and serum sFlt-1. This study suggests that celery herbs extract (CHE) has an apigenin-flavonoid compound which can prevent intrauterine growth restriction (IUGR) via suppression of antiangiogenic factor production in preeclampsia mice model.

Resveratrol mitigates trophoblast and endothelial dysfunction partly via activation of nuclear factor erythroid 2-related factor-2

IntroductionMaternal endothelial dysfunction underlying preeclampsia arises from excessive placental release of anti-angiogenic factors, such as soluble fms-like tyrosine kinase-1 (sFlt1), soluble endoglin (sEng) and activin A. Resveratrol, an activator of the nuclear factor erythroid 2-related factor-2 (Nrf2) transcription factor, mediates the gene expression of antioxidant and vasoprotective factors that may counter the endothelial damage imposed by these anti-angiogenic factors. The objective of this study was to assess whether resveratrol could reduce placental oxidative stress and production of anti-angiogenic factors in vitro and/or improve in vitro markers of endothelial dysfunction via Nrf2 activation. MethodWe used in vitro term placental explants to assess the effects of resveratrol on placental oxidative stress and production of sFlt1, sEng and activin A. Using human umbilical vein endothelial cells we investigated the effects of resveratrol on markers of in vitro endothelial dysfunction, including the expression of intercellular adhesion molecule 1 (ICAM1), vascular cell adhesion molecule 1 (VCAM1), E-selectin and endothelin-1, and endothelial permeability. To confirm that resveratrol mediated its effects via Nrf2, we examined the impact of resveratrol on the same in vitro markers of endothelial and placental dysfunction following Nrf2 knockdown. ResultsResveratrol significantly decreased placental oxidative stress and the production of sFlt1 and activin A. Resveratrol significantly mitigated tumor necrosis factor–α stimulated endothelial expression of ICAM1, VCAM1, E-selectin and endothelin-1 and prevented an increase in endothelial monolayer permeability. Nrf2 knockdown abolished some of the protective effects of resveratrol on endothelial cells, but not in primary trophoblast cells. ConclusionFeatures of placental and endothelial dysfunction characteristic of preeclampsia are improved by resveratrol in vitro, partially via the modulation of Nrf2.

Activating Transcription Factor 3 Is Reduced in Preeclamptic Placentas and Negatively Regulates sFlt-1 (Soluble fms-Like Tyrosine Kinase 1), Soluble Endoglin, and Proinflammatory Cytokines in Placenta.

Preeclampsia is a major pregnancy complication associated with poor placental perfusion and placental hypoxia. Systemic and placental inflammation and elevated placental secretion of the antiangiogenic factors sFlt-1 (soluble fms-like tyrosine kinase 1) and sEng (soluble endoglin) are hallmarks of preeclampsia, causing endothelial dysfunction and multiorgan injury. A molecule that links placental hypoxia, inflammation, and antiangiogenic factor release has not been described. ATF3 (activating transcription factor 3) is highly expressed in placenta. We assessed whether placental ATF3 is dysregulated in preterm preeclampsia, is altered by hypoxia, and regulates proinflammatory cytokine and antiangiogenic factor production. ATF3 mRNA and protein expression was significantly reduced in preterm preeclamptic placentas compared with gestation-matched controls. Hypoxia reduced ATF3 expression in primary cytotrophoblast and placental explants. Silencing ATF3 in primary cytotrophoblast increased proinflammatory cytokine (IL-6 [interleukin 6], TNF-α [tumor necrosis factor α]) and NF-κB (nuclear factor κB) expression. In silico analysis identified an ATF3-binding site in the promoter of Flt-1 (the transcript from which sFlt-1 is produced). Silencing ATF3 increased sFlt-1 and sEng secretion from primary cytotrophoblast possibly by increasing Rab11a and Arf1, cargo proteins that facilitate exosomal release of sFlt-1. ATF3 knockout mice did not have a preeclampsia phenotype, suggesting that these pathways may be specific to humans (preeclampsia is a uniquely human condition). To conclude, we have shown that ATF3 is decreased in preeclamptic placentas and that this decrease is likely to occur after prolonged hypoxia. We show that ATF3 is a regulator of placental proinflammatory cytokines and antiangiogenic factors sFlt-1 and sEng. Therefore, reduced ATF3 may be centrally involved in the pathology of preeclampsia.

The effects of very low doses of chemotherapeutic drugs on endothelial cells.

e23014 Background: Angiogenesis is one of the key processes leading tumor progression and metastasis. Conventional chemotherapeutic drugs are usually cytotoxic to tumor cells at the therapeutic doses. The lowered doses of those drugs have been shown to induce apoptosis in endothelial cells and implemented as metronomic schedule in the clinic. We aimed to study the effects of very low doses of chemotherapeutic drugs on endothelial cell functions and gene and protein expression of pro-and anti-angiogenic factors. Methods: We first determined the IC50 doses of 5-fluorouracil, irinotecan, and oxaliplatin in a human umbilical endothelial cell line (HUVEC). Then we’ve tested the effects of the IC50, -1log, -2log and -3log of IC50 doses on endothelial cell migration and tube formation. We’ve also studied the effects of the assigned doses on expression levels of the genes VEGF-A, VEGFR2, PDGF-B, angiopoietin-2 (Ang-2), endothelin-1 (End-1), thrombospondin-1 (Tbs-1), iNOS, eNOS by real-time PCR, protein secretion levels of VEGF-A, PDGF-BB, Ang-2, End-1 and Tbs-1 by ELISA and nitrate/nitrite production by a colorimetric assay. Results: The IC50 dose levels of 5-FU, irinotecan, and oxaliplatin were 9,2 μM, 9,8 μM and 11,8 μM, respectively. We found that all the -1log, -2log and -3log of IC50 doses of the drugs tested in the study significantly decreased the migration capacity of the HUVEC cells. However, no significant changes were observed in tube formation of endothelial cells. Likewise, the expression levels of pro-angiogenic genes including Ang-2, iNOS, eNOS, PDGF-B and VEGF-A were significantly decreased at the same doses of the drugs. Accordingly, the levels of Tbs-1 were significantly increased at all 3 dose levels of the drugs. The most efficacious doses of the drugs to decrease NO levels in HUVEC cells were -1logIC50 for 5-FU, -2logIC50 for irinotecan and -3logIC50 for oxaliplatin. Conclusions: In conclusion, our results suggest that 5-FU, irinotecan, and oxaliplatin could decrease the pro-angiogenic and increase anti-angiogenic factor production in endothelial cells at the doses much lower than the usual metronomic schedules and warrant further research in animal models and clinical trials as an anti-angiogenic treatment modality.

The relationship between circulating tissue transglutaminase, soluble fms-like tyrosine kinase-1, soluble endoglin and vascular endothelial growth factor in pre-eclampsia.

Preeclampsia (PE) is a pregnancy-specific syndrome that causes substantial maternal and fetal morbidity and mortality. Increased production of antiangiogenic factors, soluble fms-like tyrosine kinase receptor-1 (sFlt-1) and soluble endoglin (sEng), as well as decreased circulating levels of free vascular endothelial growth factor (VEGF), contribute to the pathophysiology of PE. Our objective was to evaluate a novel placenta-related factor, tissue transglutaminase (tTG), in PE and to investigate the correlation among tTG and sFlt-1, sEng and VEGF levels in both normotensive pregnant patients and PE patients. A total of 205 pregnant primigravid women were recruited and divided into a normotensive group (n=100), a mild PE group (n=45) and a severe PE group (n=60). Circulating serum tTG, sFlt-1, sEng and free VEGF levels were determined using an enzyme-linked immunosorbent assay. The severe PE group showed higher levels of tTG, sFlt-1 and sEng than the mild PE and normotensive groups. Free VEGF levels were lower in the severe PE group than in the mild PE and normotensive groups. tTG correlated significantly with sFlt-1, sEng and VEGF in the PE groups, whereas this correlation was not observed in the normotensive group. The tTG, sFlt-1, sEng and VEGF levels showed a significant correlation with mean arterial pressure in the PE groups but not in the normotensive group. The tTG, sFlt-1, sEng and VEGF levels correlated with the degree of proteinuria. Our results reveal that tTG is associated with sFlt-1, sEng and VEGF in the maternal circulation of PE patients, suggesting that tTG may have a role in the pathogenesis of PE.

Acute Atherosis of the Uterine Spiral Arteries: Clinicopathologic Implications.

Acute atherosis is unique vascular changes of the placenta associated with poor placentation. It is characterized by subendothelial lipid-filled foam cells, fibrinoid necrosis of the arterial wall, perivascular lymphocytic infiltration, and it is histologically similar to early-stage atherosclerosis. Acute atherosis is rare in normal pregnancies, but is frequently observed in non- transformed spiral arteries in abnormal pregnancies, such as preeclampsia, small for gestational age (SGA), fetal death, spontaneous preterm labor and preterm premature rupture of membranes. In preeclampsia, spiral arteries fail to develop physiologic transformation and retain thick walls and a narrow lumen. Failure of physiologic transformation of spiral arteries is believed to be the main cause of uteroplacental ischemia, which can lead to the production of anti-angiogenic factors and induce endothelial dysfunction and eventually predispose the pregnancy to preeclampsia. Acute atherosis is more frequently observed in the spiral arteries of the decidua of the placenta (parietalis or basalis) than in the decidual or myometrial segments of the placental bed. The presence and deeper location of acute atherosis is associated with poorer pregnancy outcomes, more severe disease, earlier onset of preeclampsia, and a greater frequency of SGA neonates in patients with preeclampsia. Moreover, the idea that the presence of acute atherosis in the placenta may increase the risk of future cardiovascular disease in women with a history of preeclampsia is of growing concern. Therefore, placental examination is crucial for retrospective investigation of pregnancy complications and outcomes, and accurate placental pathology based on universal diagnostic criteria in patients with abnormal pregnancies is essential for clinicopathologic correlation.

Abstract 4071: Cancer stem cell-induced vascularization of skin cancer tumors

Abstract Our previous studies identify a subpopulation of epidermal cancer stem (ECS) cells in human squamous cell carcinoma that are efficient tumor formers. These cells display enhanced invasion and migration, and form large, invasive and highly aggressive tumors. We now show that ECS cells produce highly vascularized tumors. This is confirmed visually and by detection of increased expression of the CD31 endothelial cell marker. To understand the molecular basis for increased vascularization, we assayed for ECS cell and non-stem cancer cell production of angiogenic factors. Pro-angiogenic factors, including VEGF-A, HIF-1α and MMP9, are produced at high level in ECS cell-derived tumors, as compared to non-stem cancer cell tumors. In addition, anti-angiogenic factor production (thrombospondin-1, endothelin-1, serpin B5) is reduced. This is consistent with the finding that ECS cell tumor extracts more efficiently stimulate HUVEC cell tube formation. Moreover, these ECS tumor-derived cells display enhanced spheroid formation, and cell migration and invasion. Knockdown of VEGF-A or VEGFR2, or treatment with anti-VEGF-A attenuates these events. In addition, treatment with bevacizumab (anti-VEGF-A) suppresses tumor formation in vivo. These studies suggest that the VEGF-A signaling pathway is enhanced in ECS cells and is in part responsible the ability of these cells to form aggressive and highly vascularized tumors. Citation Format: Dan Grun, Gautam Adhikary, Richard Eckert. Cancer stem cell-induced vascularization of skin cancer tumors. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4071. doi:10.1158/1538-7445.AM2015-4071

Transthyretin Is Dysregulated in Preeclampsia, and Its Native Form Prevents the Onset of Disease in a Preclinical Mouse Model

Preeclampsia is a major pregnancy complication with potential short- and long-term consequences for both mother and fetus. Understanding its pathogenesis and causative biomarkers is likely to yield insights for prediction and treatment. Herein, we provide evidence that transthyretin, a transporter of thyroxine and retinol, is aggregated in preeclampsia and is present at reduced levels in sera of preeclamptic women, as detected by proteomic screen. We demonstrate that transthyretin aggregates form deposits in preeclampsia placental tissue and cause apoptosis. By using in vitro approaches and a humanized mouse model, we provide evidence for a causal link between dysregulated transthyretin and preeclampsia. Native transthyretin inhibits all preeclampsia-like features in the humanized mouse model, including new-onset proteinuria, increased blood pressure, glomerular endotheliosis, and production of anti-angiogenic factors. Our findings suggest that a focus on transthyretin structure and function is a novel strategy to understand and combat preeclampsia.