- Email: [email protected]
AXL signaling induces preeclampsia
Abstracts / Placenta 45 (2016) 63e133
for presence of VEGF+936C/T, sFlt-1(+4244G/A, -523C/G, -4771G/T) polymorphisms. Serum levels of VEGF-A and sFlt-1 were measured by sandwich ELISA. Results: Increased frequency of CT genotype of VEGF+936C/T, GA genotype of sFlt-1+4244G/A and CG genotype of sFlt-1-523C/G was seen in patients compared to controls. ELISA results showed lower VEGF-A (214.19pg/ml vs 225.65pg/ml) and higher sFlt-1 levels (2932.81pg/ml vs 1114.94; p<0.05) in patients compared to controls. Correlation of genotypes with protein levels showed that pregnant women with VEGF+936CT genotype had lower levels of VEGF-A and sFlt-1+4244GA genotype had increased serum levels of sFlt-1. Conclusion: The pilot study shows for the first time a possible association of VEGF and sFlt-1 polymorphisms with gene expression leading to altered protein levels in preeclamptic patients of Indian origin. P2.52 GROWTH ARREST-SPECIFIC PREECLAMPSIA
6
(GAS6)/AXL
SIGNALING
INDUCES
119
screening completed at 6 months postpartum were selected (n¼48). Blinded pathology reports were reviewed for lesions of maternal malperfusion, decidual vasculopathy, fetoplacental thrombotic vasculopathy, chronic uteroplacental separation, maternal-fetal interface disturbance and chronic inflammation. The number of lesions reported between high and low risk women was compared. Results: Of the 48 women, 32 (67%) were classified as high risk for lifetime CVD at 6 months postpartum. Approximately half of the placentas in each group were histologically normal (no lesions observed). Lesions of maternal underperfusion, acute atherosis and evidence of fetal thrombotic vasculopathy were more common in the high risk group although differences were non-significant (Fig. 1). Conclusion: Placental lesions may differentiate women deemed high risk for CVD following a pregnancy complicated by PE but larger numbers are needed to confirm these trends. Placental pathology may offer a unique modality for identifying women who should receive CVD follow-up postpartum.
Camilo Mejia, Montana Wayment, Troy Monson, Paul Reynolds, Juan Arroyo. Brigham Young University, Provo, UT, USA Objectives: Preeclampsia (PE) is a complicated obstetric complication characterized by increased blood pressure and decreased trophoblast invasion. The growth arrest-specific 6 (Gas6) protein is known to induce dynamic cellular responses including mechanisms of apoptosis, cell migration, and invasion. Gas6 binds and induces signaling through the AXL receptor tyrosine kinase and its expression is elevated in PE suggesting a mechanistic role for its signaling pathway during disease progression. Our objective was to identify the in vivo effects of increased Gas6 protein during pregnancy. Methods: Holtzman pregnant rats were treated daily with 3ug/ml of Gas6 for 11 days from day 7 to day 17 of gestation (dGA). At the time of necropsy (18 dGA) placental and fetal weights were recorded, proteinuria determined and tissues were immediately isolated for histological, RNA and protein analysis. First trimester trophoblast cells were treated with Gas6 and invasion was measured using an xCELLigence RTCA DP (Real Time Cell Analysis Dual Plate) instrument. Results: In Gas6 treated rats, we observed: 1) increased systolic and diastolic blood pressure; 2) increased proteinuria; 3) decreased trophoblast invasion 4) no differences in placental or fetal weights, 5) increased placental cleaved caspase 3 (80%) and phospho AXL (44%) at term, 6) and decreased Slug protein (27%). In vitro work demonstrated decreased trophoblast invasion after 24 hours of Gas6 treatment. Protein and PCR arrays further implicated a host of signaling intermediates in Gas6/AXLmediated PE. Conclusions: We conclude that Gas6/AXL interactions are involved in the induction of PE symptoms and this interaction directly inhibits trophoblast invasion coincident with PE. These results provide insight into pathways associated with increased Gas6 during PE and could be useful in the clarification of potential therapeutic mechanisms. P2.53 CAN PLACENTAL PATHOLOGY IDENTIFY WOMEN AT HIGH RISK OF CARDIOVASCULAR DISEASE FOLLOWING PREECLAMPSIA? Samantha Benton 1, David Grynspan 2, Graeme Smith 3, Shannon Bainbridge 1. 1 University of Ottawa, Ottawa, Ontario, Canada; 2 Children's Hospital of Eastern Ontario, Ottawa, Ontario, Canada; 3 Queen's University, Kingston, Ontario, Canada Objectives: Pre-eclampsia (PE) is a significant risk factor for cardiovascular disease (CVD) in later life. However, we currently have no easy method to identify which women are at highest risk at the time of delivery and who would benefit most from cardiovascular follow-up postpartum. Placenta pathology, a routine clinical examination conducted in the context of PE, may provide a tool to identify these women. In this pilot study, we investigated placental lesions in women deemed high and low risk for lifetime CVD in postpartum follow-up. Methods: From the Maternal Health Clinic (Kingston General Hospital), women with placental pathology performed at delivery and CVD risk
Fig. 1. Placental lesions observed in high and low risk groups.Ă
P2.54 PIM 1KINASE IN PREECLAMPSIA Stella M. Mary 1, Maja Weber 1, Wittaya Chaiwangyen 1, 2, Ekkehard Schleussner 1, Udo R. Markert 1. 1 Placenta-Lab, Department of Obstetrics, University hospital Jena, Jena, Germany; 2 Department of Biochemistry, School of Medical Sciences, University of Phayao, Phayao, Thailand Background: Preeclampsia is a pregnancy pathology which involves high blood pressure, proteinuria and inflammation with an onset in the second half of pregnancy. The proviral insertion in murine (PIM) lymphoma proteins are a serine /threonine kinase family composed of three isoforms PIM1, PIM2 and PIM3. PIM kinases play a critical role in the control of cell survival, proliferation, homing, migration. So far PIM kinases have not been studied in pregnancy related pathologies. The aim of this study was to investigate the expression of PIM kinases in placentae from patients with preeclampsia. Methods: Placental tissues were obtained from five preeclampsia patients and five healthy controls immediately after delivery, fixed in formaldehyde and embedded in paraffin. Immunohistochemistry staining for PIM1 was performed to investigate its expression and local distribution. We have applied an immune reactive score for the analysis. Results: PIM1 kinase was expressed in the cytoplasma of syncytiotrophoblast and in extra villous trophoblast cells. Expression was lower in preeclampsia when compared to healthy tissues.
for presence of VEGF+936C/T, sFlt-1(+4244G/A, -523C/G, -4771G/T) polymorphisms. Serum levels of VEGF-A and sFlt-1 were measured by sandwich ELISA. Results: Increased frequency of CT genotype of VEGF+936C/T, GA genotype of sFlt-1+4244G/A and CG genotype of sFlt-1-523C/G was seen in patients compared to controls. ELISA results showed lower VEGF-A (214.19pg/ml vs 225.65pg/ml) and higher sFlt-1 levels (2932.81pg/ml vs 1114.94; p<0.05) in patients compared to controls. Correlation of genotypes with protein levels showed that pregnant women with VEGF+936CT genotype had lower levels of VEGF-A and sFlt-1+4244GA genotype had increased serum levels of sFlt-1. Conclusion: The pilot study shows for the first time a possible association of VEGF and sFlt-1 polymorphisms with gene expression leading to altered protein levels in preeclamptic patients of Indian origin. P2.52 GROWTH ARREST-SPECIFIC PREECLAMPSIA
6
(GAS6)/AXL
SIGNALING
INDUCES
119
screening completed at 6 months postpartum were selected (n¼48). Blinded pathology reports were reviewed for lesions of maternal malperfusion, decidual vasculopathy, fetoplacental thrombotic vasculopathy, chronic uteroplacental separation, maternal-fetal interface disturbance and chronic inflammation. The number of lesions reported between high and low risk women was compared. Results: Of the 48 women, 32 (67%) were classified as high risk for lifetime CVD at 6 months postpartum. Approximately half of the placentas in each group were histologically normal (no lesions observed). Lesions of maternal underperfusion, acute atherosis and evidence of fetal thrombotic vasculopathy were more common in the high risk group although differences were non-significant (Fig. 1). Conclusion: Placental lesions may differentiate women deemed high risk for CVD following a pregnancy complicated by PE but larger numbers are needed to confirm these trends. Placental pathology may offer a unique modality for identifying women who should receive CVD follow-up postpartum.
Camilo Mejia, Montana Wayment, Troy Monson, Paul Reynolds, Juan Arroyo. Brigham Young University, Provo, UT, USA Objectives: Preeclampsia (PE) is a complicated obstetric complication characterized by increased blood pressure and decreased trophoblast invasion. The growth arrest-specific 6 (Gas6) protein is known to induce dynamic cellular responses including mechanisms of apoptosis, cell migration, and invasion. Gas6 binds and induces signaling through the AXL receptor tyrosine kinase and its expression is elevated in PE suggesting a mechanistic role for its signaling pathway during disease progression. Our objective was to identify the in vivo effects of increased Gas6 protein during pregnancy. Methods: Holtzman pregnant rats were treated daily with 3ug/ml of Gas6 for 11 days from day 7 to day 17 of gestation (dGA). At the time of necropsy (18 dGA) placental and fetal weights were recorded, proteinuria determined and tissues were immediately isolated for histological, RNA and protein analysis. First trimester trophoblast cells were treated with Gas6 and invasion was measured using an xCELLigence RTCA DP (Real Time Cell Analysis Dual Plate) instrument. Results: In Gas6 treated rats, we observed: 1) increased systolic and diastolic blood pressure; 2) increased proteinuria; 3) decreased trophoblast invasion 4) no differences in placental or fetal weights, 5) increased placental cleaved caspase 3 (80%) and phospho AXL (44%) at term, 6) and decreased Slug protein (27%). In vitro work demonstrated decreased trophoblast invasion after 24 hours of Gas6 treatment. Protein and PCR arrays further implicated a host of signaling intermediates in Gas6/AXLmediated PE. Conclusions: We conclude that Gas6/AXL interactions are involved in the induction of PE symptoms and this interaction directly inhibits trophoblast invasion coincident with PE. These results provide insight into pathways associated with increased Gas6 during PE and could be useful in the clarification of potential therapeutic mechanisms. P2.53 CAN PLACENTAL PATHOLOGY IDENTIFY WOMEN AT HIGH RISK OF CARDIOVASCULAR DISEASE FOLLOWING PREECLAMPSIA? Samantha Benton 1, David Grynspan 2, Graeme Smith 3, Shannon Bainbridge 1. 1 University of Ottawa, Ottawa, Ontario, Canada; 2 Children's Hospital of Eastern Ontario, Ottawa, Ontario, Canada; 3 Queen's University, Kingston, Ontario, Canada Objectives: Pre-eclampsia (PE) is a significant risk factor for cardiovascular disease (CVD) in later life. However, we currently have no easy method to identify which women are at highest risk at the time of delivery and who would benefit most from cardiovascular follow-up postpartum. Placenta pathology, a routine clinical examination conducted in the context of PE, may provide a tool to identify these women. In this pilot study, we investigated placental lesions in women deemed high and low risk for lifetime CVD in postpartum follow-up. Methods: From the Maternal Health Clinic (Kingston General Hospital), women with placental pathology performed at delivery and CVD risk
Fig. 1. Placental lesions observed in high and low risk groups.Ă
P2.54 PIM 1KINASE IN PREECLAMPSIA Stella M. Mary 1, Maja Weber 1, Wittaya Chaiwangyen 1, 2, Ekkehard Schleussner 1, Udo R. Markert 1. 1 Placenta-Lab, Department of Obstetrics, University hospital Jena, Jena, Germany; 2 Department of Biochemistry, School of Medical Sciences, University of Phayao, Phayao, Thailand Background: Preeclampsia is a pregnancy pathology which involves high blood pressure, proteinuria and inflammation with an onset in the second half of pregnancy. The proviral insertion in murine (PIM) lymphoma proteins are a serine /threonine kinase family composed of three isoforms PIM1, PIM2 and PIM3. PIM kinases play a critical role in the control of cell survival, proliferation, homing, migration. So far PIM kinases have not been studied in pregnancy related pathologies. The aim of this study was to investigate the expression of PIM kinases in placentae from patients with preeclampsia. Methods: Placental tissues were obtained from five preeclampsia patients and five healthy controls immediately after delivery, fixed in formaldehyde and embedded in paraffin. Immunohistochemistry staining for PIM1 was performed to investigate its expression and local distribution. We have applied an immune reactive score for the analysis. Results: PIM1 kinase was expressed in the cytoplasma of syncytiotrophoblast and in extra villous trophoblast cells. Expression was lower in preeclampsia when compared to healthy tissues.