- Email: [email protected]
P6. Active and inactivated hemopexin in pregnancy and preeclampsia: Do they activate monocytes and endothelial cells?
Poster Presentations / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 1 (2011) 273–299
P5. Bioinformatic analysis of protein–protein interaction network in preeclampsia E. Tejera a, J. Bernardes b, I. Rebelo a (a Faculty of Pharmacy/IBMC, Porto, Portugal, b Faculty of Medicine/S João Hospital, Porto, Portugal) Preeclampsia has been studied for many years, but until now the underlying mechanisms remain unknown. Likewise, it has not been identified a predictive diagnostic marker or an effective treatment to prolong the fetus and/or mother wellbeing. Although there are many articles about preeclampsia few are related with bioinformatic analysis. The objective of present study is to analyze the protein–protein interaction network of preeclampsia genes/proteins in order to identify significant biological process and metabolic functions as well as central genes/proteins in preeclampsia. An extensive collection of genes/proteins involved in preeclampsia was obtained and analyzed. A genes/proteins dataset (645) of significantly differentiated regulation/ expression between normal and preeclampsia was built considering both public proteomic data and publication analysis using several tools. After a mapping of the dataset under several public protein/genes database a protein–protein interaction network was developed with a total of 8046 located interactions. Both, the dataset and the interaction network were analyzed to identify statistical significant gene ontology process and functions as well as metabolic pathways. The results reveal a significant modification of biological process related with immunological, lipid and several enzymatic processes. However some specific proteins present a wide type of metabolic involvement reflecting the complexity and multifactorial manifestation of preeclampsia. Topic of the abstract: Placental and fetal preeclampsia. First Author < 35 years doi:10.1016/j.preghy.2011.08.066
P6. Active and inactivated hemopexin in pregnancy and preeclampsia: Do they activate monocytes and endothelial cells? F. Spaans, C. Chiang, W.W. Bakker, T. Borghuis, M.M. Faas (Div. of Medical Biology and Dept. Pathology and Medical Biology, University Medical Center Groningen, Groningen, The Netherlands) Introduction: Normal pregnancy is characterized by activation of monocytes, and this is exacerbated in preeclampsia. In addition, activation of endothelial cells is also observed in preeclampsia. The mechanism of activation of monocytes and endothelial cells remains unknown. Hemopexin (Hx) activity increases during normal pregnancy, which facilitates reduced Angiotensin II sensitivity and normal expansion of the vascular bed. However, in preeclampsia Hx activity is not elevated and this is thought to be due to high extracellular ATP titers in preeclamptic patients, which can inhibit Hx activity. In this study we aimed to investigate the effect of extracellular ATP, active and ATP inactivated Hx
275
on activation of human monocytes and endothelial cells in vitro. Methods: Monomac-6 cells (human monocyte cell line) and Human Umbillical Vein Endothelial Cells (HUVECs) were separately stimulated with LPS (2 lg/ml, positive control), ATP (250 lmol/L), active Hx (100 lg/ml, reflecting the normal pregnant situation), or Hx inhibited by ATP (Hx-ATP, 100 lg/ ml, reflecting the situation in preeclampsia) and compared to unstimulated control. After 4 h stimulation the cells were stained for ICAM-1 expression to determine the activation status using flow cytometry. Results: Stimulation of monocytes and endothelial cells with either LPS, Hx or Hx-ATP increases ICAM-1 expression (p < 0.05). ATP stimulation alone did not change ICAM-1 expression on both cell types. On monocytes ICAM-1 expression was higher after stimulation with Hx-ATP compared to stimulation with active Hx (see Fig. 1, *p < 0.05; **p < 0.01 Mann–Whitney U test). Conclusion/discussion: Human monocytes and endothelial cells become activated when stimulated with active Hx, which is in line with the moderate inflammation that is observed in normal pregnancy. However, Hx-ATP activates monocytes even more, resembling what is observed in preeclampsia. These data suggest a possible role for Hx-ATP in inducing the inflammatory response in preeclampsia. Topic of the abstract: Placental and fetal preeclampsia. First Author < 35 years: Yes. doi:10.1016/j.preghy.2011.08.067
P7. Role of EGF-like domain 7 (Egfl7) in placental development and implantation Silvia Salvi a, Sergio Ferrazzani a, Lucia Vecchione b, Gregorio Siracusa b, Heidi Stuhlmann c, Luisa Campagnolo b (a Catholic University of Sacred Heart, Rome, Italy, b University of Rome ‘‘Tor Vergata’’, Rome, Italy, c Weill Cornell Medical College, New York, USA) Objectives: Implantation and placental development are essential for the patterning of the embryo, for its growth and survival upon implantation, as well as for preserving the health of the fetus and mother. Inadequate invasion of trophoblast cells into the maternal decidua and the deficient growth of fetal blood vessels are actually recognized as possible pathogenetic factors of preeclampsia (PE). To examine molecular mechanisms that control these events, we are focusing in our studies on the endothelial gene Egfl7. Our preliminary studies show that Egfl7 has a critical role in molecular processes that control human vascular development and proper placental implantation: in particular, Egfl7 is expressed in the trophectoderm and in the fetal placental vasculature. To assess the involvement of Egfl7 in the development of PE, we investigate the EGFL7 expression patterns between PE and control placentas. Materials and methods: Two study groups are considered: (1) a first of women with pregnancies complicated by early onset PE ; (2) a second study group of healthy women. The placental biopsies were obtained immediately after caesarean section from both groups, the PE patients and the
P5. Bioinformatic analysis of protein–protein interaction network in preeclampsia E. Tejera a, J. Bernardes b, I. Rebelo a (a Faculty of Pharmacy/IBMC, Porto, Portugal, b Faculty of Medicine/S João Hospital, Porto, Portugal) Preeclampsia has been studied for many years, but until now the underlying mechanisms remain unknown. Likewise, it has not been identified a predictive diagnostic marker or an effective treatment to prolong the fetus and/or mother wellbeing. Although there are many articles about preeclampsia few are related with bioinformatic analysis. The objective of present study is to analyze the protein–protein interaction network of preeclampsia genes/proteins in order to identify significant biological process and metabolic functions as well as central genes/proteins in preeclampsia. An extensive collection of genes/proteins involved in preeclampsia was obtained and analyzed. A genes/proteins dataset (645) of significantly differentiated regulation/ expression between normal and preeclampsia was built considering both public proteomic data and publication analysis using several tools. After a mapping of the dataset under several public protein/genes database a protein–protein interaction network was developed with a total of 8046 located interactions. Both, the dataset and the interaction network were analyzed to identify statistical significant gene ontology process and functions as well as metabolic pathways. The results reveal a significant modification of biological process related with immunological, lipid and several enzymatic processes. However some specific proteins present a wide type of metabolic involvement reflecting the complexity and multifactorial manifestation of preeclampsia. Topic of the abstract: Placental and fetal preeclampsia. First Author < 35 years doi:10.1016/j.preghy.2011.08.066
P6. Active and inactivated hemopexin in pregnancy and preeclampsia: Do they activate monocytes and endothelial cells? F. Spaans, C. Chiang, W.W. Bakker, T. Borghuis, M.M. Faas (Div. of Medical Biology and Dept. Pathology and Medical Biology, University Medical Center Groningen, Groningen, The Netherlands) Introduction: Normal pregnancy is characterized by activation of monocytes, and this is exacerbated in preeclampsia. In addition, activation of endothelial cells is also observed in preeclampsia. The mechanism of activation of monocytes and endothelial cells remains unknown. Hemopexin (Hx) activity increases during normal pregnancy, which facilitates reduced Angiotensin II sensitivity and normal expansion of the vascular bed. However, in preeclampsia Hx activity is not elevated and this is thought to be due to high extracellular ATP titers in preeclamptic patients, which can inhibit Hx activity. In this study we aimed to investigate the effect of extracellular ATP, active and ATP inactivated Hx
275
on activation of human monocytes and endothelial cells in vitro. Methods: Monomac-6 cells (human monocyte cell line) and Human Umbillical Vein Endothelial Cells (HUVECs) were separately stimulated with LPS (2 lg/ml, positive control), ATP (250 lmol/L), active Hx (100 lg/ml, reflecting the normal pregnant situation), or Hx inhibited by ATP (Hx-ATP, 100 lg/ ml, reflecting the situation in preeclampsia) and compared to unstimulated control. After 4 h stimulation the cells were stained for ICAM-1 expression to determine the activation status using flow cytometry. Results: Stimulation of monocytes and endothelial cells with either LPS, Hx or Hx-ATP increases ICAM-1 expression (p < 0.05). ATP stimulation alone did not change ICAM-1 expression on both cell types. On monocytes ICAM-1 expression was higher after stimulation with Hx-ATP compared to stimulation with active Hx (see Fig. 1, *p < 0.05; **p < 0.01 Mann–Whitney U test). Conclusion/discussion: Human monocytes and endothelial cells become activated when stimulated with active Hx, which is in line with the moderate inflammation that is observed in normal pregnancy. However, Hx-ATP activates monocytes even more, resembling what is observed in preeclampsia. These data suggest a possible role for Hx-ATP in inducing the inflammatory response in preeclampsia. Topic of the abstract: Placental and fetal preeclampsia. First Author < 35 years: Yes. doi:10.1016/j.preghy.2011.08.067
P7. Role of EGF-like domain 7 (Egfl7) in placental development and implantation Silvia Salvi a, Sergio Ferrazzani a, Lucia Vecchione b, Gregorio Siracusa b, Heidi Stuhlmann c, Luisa Campagnolo b (a Catholic University of Sacred Heart, Rome, Italy, b University of Rome ‘‘Tor Vergata’’, Rome, Italy, c Weill Cornell Medical College, New York, USA) Objectives: Implantation and placental development are essential for the patterning of the embryo, for its growth and survival upon implantation, as well as for preserving the health of the fetus and mother. Inadequate invasion of trophoblast cells into the maternal decidua and the deficient growth of fetal blood vessels are actually recognized as possible pathogenetic factors of preeclampsia (PE). To examine molecular mechanisms that control these events, we are focusing in our studies on the endothelial gene Egfl7. Our preliminary studies show that Egfl7 has a critical role in molecular processes that control human vascular development and proper placental implantation: in particular, Egfl7 is expressed in the trophectoderm and in the fetal placental vasculature. To assess the involvement of Egfl7 in the development of PE, we investigate the EGFL7 expression patterns between PE and control placentas. Materials and methods: Two study groups are considered: (1) a first of women with pregnancies complicated by early onset PE ; (2) a second study group of healthy women. The placental biopsies were obtained immediately after caesarean section from both groups, the PE patients and the