- Email: [email protected]
[62-OR]
32
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 5 (2015) 2–52
Disclosures: Z.A. Brown: None. S. Schalekamp-Timmermans: None. A. Hofman: None. V. Jaddoe: None. E. Steegers: None.
doi:10.1016/j.preghy.2014.10.064
[61-OR] Functional regulation of trophoblast cells by lysophosphatidic acid signaling and its pathologic relevance to PIH Mayuko Ichikawa, Takeshi Nagamatsu, Tatsuya Fujii, Mari Hoya, Yuki Kawai-Iwasawa, Katsutoshi Oda, Kei Kawana, Takahiro Yamashita, Yutaka Osuga, Tomoyuki Fujii (The University of Tokyo, Tokyo, Japan) Objectives: Lysophosphatidic acid (LPA) is a new class lipid mediator and exerts varied physiological and pathological functions through the binding to specific cell-membrane receptors. Impaired implantation was reported in mice deficient in LPA3, one of LPA receptors, suggesting the critical role of this pathway in reproduction. A secreting protein, autotaxin (ATX) is a key enzyme to determine local LPA production. We aimed to explore the regulatory role of trophoblast cell function by ATX-LPA-LAP3 pathway and its association with the pathology of pregnancy induced hypertension (PIH). Methods: (1) We analyzed the local expression of ATX and LPA3 in human placenta using immunohistochemistry. (2) We transfected LPA3 gene to HTR-8/SVneo, a trophoblastic cell line, and stimulated it with the specific agonist of LPA3. Real-time PCR was conducted to examined the impact of the stimulation on the expression level of angiogenic factors in LPA3-transfected cells. (3) We analyzed the difference in ATX mRNA expression in human placenta between normal pregnancy and PIH. All placental samples and the experiments were conducted under the approval of the ethical committee in our facility. Results: (1) ATX-specific staining is confirmed in all types of trophoblast cells. The expression of LPA3 was restricted to differentiated trophoblasts including syncytiotrophoblast and extra-villous trophoblasts. (2) LPA3-transfected HTR-8/SVneo enhanced cyclooxygenase-2 (COX-2), interleukin-8 (IL-8), and vascular endotherial growth factor(VEGF) expression remarkably upon the stimulation with a specific agonist to LPA3. (3) In real-time PCR analysis, ATX mRNA expression level was significantly lower in the placentas with PIH compared with normal placentas.
Conclusions: Our findings revealed that the ATX-LPA-LPA3 pathway regulates the expression of angiogenic factors in trophoblasts. Reduced ATX production occurring in preeclamptic placentas might diminish the angiogenic property in trophoblasts, consequently leading to aberrant placentation. Disclosures: M. Ichikawa: None. T. Nagamatsu: None. T. Fujii: None. M. Hoya: None. Y. Kawai-Iwasawa: None. K. Oda: None. K. Kawana: None. T. Yamashita: None. Y. Osuga: None. T. Fujii: None. doi:10.1016/j.preghy.2014.10.065
[62-OR] Impaired autophagy in placenta of pregnant women with preeclampsia Ingrid C. Weel a, Vanessa R. Ribeiro a, Mariana Romao a, Mariana L. Matias b, Vera T. M Borges b, Jose C. Peracoli b, Joao P. Araujo Jr a, Maria T. S Peracoli a (a Institute of Biosciences, Sao Paulo State University, Botucatu, SP, Brazil, b Botucatu Medical School, Sao Paulo State University, Botucatu, SP, Brazil) Objectives: Autophagy is a lysosomal degradation pathway that removes protein aggregates and damaged organelles maintaining the cellular integrity. It is an inducible process that responds to the environment and it seems to be essential for cell survival during stress, starvation, hypoxia and consequently to the placenta implantation and development. Studies on the role of autophagy in placenta from pregnant women with preeclampsia are scarce and still unclear. The aim of this study was to compare the expression of autophagy-related proteins, LC3-II, beclin-1 as well as mTOR protein in placental tissue from pregnant women with preeclampsia and normotensive pregnant women. Methods: The expression of LC3-II and beclin-1 proteins involved in the autophagosome formation as well as mTOR protein was evaluated in 18 placentas of pregnant women with preeclampsia and in 20 of normotensive pregnant women who had elective cesarean delivery before the onset of labor. Fragments of the placenta were obtained immediately after delivery and the samples were prepared for real-time quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry and transmission electron microscopy analysis. Results: LC3-II expression as detected by RT-qPCR was significant higher in placentas of normotensive pregnant women compared with preeclamptic women. No significant difference in beclin-1 and mTOR expression was seen between these two groups. LC3-II, beclin-1 and mTOR proteins were detected by immunohistochemistry in syncytiotrophoblast and cytotrophoblast in both groups, and the presence of autophagic vacuoles containing intracytoplasmatic organelles was confirmed with electron microscopy. Conclusions: The results of the impaired autophagy demonstrated by decrease of LC3- II expression in placenta of
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 5 (2015) 2–52
preeclamptic patients might play a role in the pathophysiology of preeclampsia. Financial support: FAPESP 2013/00535-1 and 2012/ 24697-8. Disclosures: I.C. Weel: None. V.R. Ribeiro: None. M. Romao: None. M.L. Matias: None. V.T. Borges: None. J.C. Peracoli: None. J.P. Araujo Jr: None. M.T. Peracoli: None.
33
indices seem to be good ultrasonographic indicators for risk estimation and prevention of complications in pregnancies with hypertension. Disclosures: Á. Altorjay: None. A. Surányi: None. M. Jakó: None. T. Nyári: None. G. Németh: None. A. Pál: None. doi:10.1016/j.preghy.2014.10.067
doi:10.1016/j.preghy.2014.10.066
[64-OR] [63-OR] Effect of pregnancies complicated by essential as well as pregnancy induced hypertension on placental vascularization Ábel Altorjay a, Andrea Surányi a, Mária Jakó a, Tibor Nyári b, Gábor Németh a, Attila Pál a (a University of Szeged, Faculty of Medicine, Szeged, Hungary, b University of Szeged, Faculty of Physics and Informatics, Szeged, Hungary) Objectives: Proper uterine and placental vascularization is crucial for the adequate development of pregnancies. Pathological fetoplacental circulation leads to elevated resistance in placental vascularization which causes insufficiency and decreased fetal oxygenation. The aim of the study was to analyze placental vascularization in pregnant women with essential hypertension (EH) and pregnancy induced hypertension (PIH) with the help of VOCAL technique. Methods: We measured placental 3-dimensional power Doppler indices such as vascularization index (VI), flow index (FI) and vascularization flow index (VFI) in pregnancies complicated by PIH (NPIH = 40), and EH (NEH = 40) and compared these to pregnancies with normal blood pressure (NNBP = 316). We analyzed the correlation between the above mentioned indices, the volume of the placenta and placental localization. We used two-sample Wilcoxon rank-sum test, Levene’s test and quantile regression. Results: VI and VFI were significantly lower in pregnancies with PIH (VImean ± SD: 10.60 ± 10.40; VFImean ± SD: 2.77 ± 2.13) than in pregnancies with NBP (p < 0.001) and they were also significantly lower in case of PIH compared to EH (p < 0.01). We haven’t found significant difference between EH (VImean ± SD: 15.46 ± 8.49; VFImean ± SD: 7.55 ± 5.06) and NBP groups. FI was significantly lower in pregnancies with EH and PIH than in pregnancies with NBP (p < 0.001), and we found significant difference between EH (FImean ±SD: 43.89 ± 9.99) and PIH (FImean ±SD: 36.42 ± 13.67) as well. The placental volume was significantly smaller (zscore: 1.5) in EH as well as PIH than in NBP pregnancies, but there was no significant difference (zscore: 0.12) between the two affected groups. The placental localization had no significant influence on VI, FI or VFI (p < 0.01). There was no correlation found between the indices and gestational weeks. Conclusions: The direct 3-dimensional measurement of placental vascularization should gather more and more ground, because anomalies of uteroplacental and fetal vascularization appear too late, when complications have already been developed. Lower placental vascularization
Molecular basis for racial disparity in pre-eclampsia Elena R. Rivers a, Anthony J. Horton a, Christopher D. Clark a, Elizabeth G. Favre a, Katherine M. Senf a, Angela F. Hawk a, Eugene Y. Chang a, Christopher J. Robinson b, Kyu-Ho Lee a (a Medical University of South Carolina, b Charleston, SC, USA, University of Virginia, Charlottesville, VA, USA) Objectives: Pre-eclampsia (PE) affects 5-8% of pregnancies, and factors causing PE and mediating observed racial disparity are poorly understood. The cardiovascular gene Nkx2-5 is expressed in trophoblast cells and abnormal amnions are observed in mice Nkx2-5 mutant mice. We assessed Nkx25-related gene expression in placenta from normal and early onset and severe pre-eclampsia (EOSPE) pregnancies. Methods: IHC and qPCR assay of Nkx2.5 and target gene expression in normal term and EOSPE placenta. Results: Nkx2-5 is highly expressed in a subset of early EOSPE placentae, and is highly correlated with expression of the PE marker sFlt-1, and with an RNA splicing factor, Sam68. These correlations are significant in Caucasian, but not African American women. siRNA knockdown of Sam68 significantly impacts sFlt-1 expression in vitro, supporting a hypothesis that Nkx2-5 impacts EOSPE severity via upregulation of Sam68 and increased sFlt-1 expression. Other Nkx2-5 targets related to metabolic stress responses are also differentially activated in between Caucasians and African American women with EOSPE. Conclusions: Nkx2-5 may play a direct role in the genesis of PE via regulation of Sam68/sFlt-1, and serve as a common link to two other stress-related modifiers of EOSPE.
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 5 (2015) 2–52
Disclosures: Z.A. Brown: None. S. Schalekamp-Timmermans: None. A. Hofman: None. V. Jaddoe: None. E. Steegers: None.
doi:10.1016/j.preghy.2014.10.064
[61-OR] Functional regulation of trophoblast cells by lysophosphatidic acid signaling and its pathologic relevance to PIH Mayuko Ichikawa, Takeshi Nagamatsu, Tatsuya Fujii, Mari Hoya, Yuki Kawai-Iwasawa, Katsutoshi Oda, Kei Kawana, Takahiro Yamashita, Yutaka Osuga, Tomoyuki Fujii (The University of Tokyo, Tokyo, Japan) Objectives: Lysophosphatidic acid (LPA) is a new class lipid mediator and exerts varied physiological and pathological functions through the binding to specific cell-membrane receptors. Impaired implantation was reported in mice deficient in LPA3, one of LPA receptors, suggesting the critical role of this pathway in reproduction. A secreting protein, autotaxin (ATX) is a key enzyme to determine local LPA production. We aimed to explore the regulatory role of trophoblast cell function by ATX-LPA-LAP3 pathway and its association with the pathology of pregnancy induced hypertension (PIH). Methods: (1) We analyzed the local expression of ATX and LPA3 in human placenta using immunohistochemistry. (2) We transfected LPA3 gene to HTR-8/SVneo, a trophoblastic cell line, and stimulated it with the specific agonist of LPA3. Real-time PCR was conducted to examined the impact of the stimulation on the expression level of angiogenic factors in LPA3-transfected cells. (3) We analyzed the difference in ATX mRNA expression in human placenta between normal pregnancy and PIH. All placental samples and the experiments were conducted under the approval of the ethical committee in our facility. Results: (1) ATX-specific staining is confirmed in all types of trophoblast cells. The expression of LPA3 was restricted to differentiated trophoblasts including syncytiotrophoblast and extra-villous trophoblasts. (2) LPA3-transfected HTR-8/SVneo enhanced cyclooxygenase-2 (COX-2), interleukin-8 (IL-8), and vascular endotherial growth factor(VEGF) expression remarkably upon the stimulation with a specific agonist to LPA3. (3) In real-time PCR analysis, ATX mRNA expression level was significantly lower in the placentas with PIH compared with normal placentas.
Conclusions: Our findings revealed that the ATX-LPA-LPA3 pathway regulates the expression of angiogenic factors in trophoblasts. Reduced ATX production occurring in preeclamptic placentas might diminish the angiogenic property in trophoblasts, consequently leading to aberrant placentation. Disclosures: M. Ichikawa: None. T. Nagamatsu: None. T. Fujii: None. M. Hoya: None. Y. Kawai-Iwasawa: None. K. Oda: None. K. Kawana: None. T. Yamashita: None. Y. Osuga: None. T. Fujii: None. doi:10.1016/j.preghy.2014.10.065
[62-OR] Impaired autophagy in placenta of pregnant women with preeclampsia Ingrid C. Weel a, Vanessa R. Ribeiro a, Mariana Romao a, Mariana L. Matias b, Vera T. M Borges b, Jose C. Peracoli b, Joao P. Araujo Jr a, Maria T. S Peracoli a (a Institute of Biosciences, Sao Paulo State University, Botucatu, SP, Brazil, b Botucatu Medical School, Sao Paulo State University, Botucatu, SP, Brazil) Objectives: Autophagy is a lysosomal degradation pathway that removes protein aggregates and damaged organelles maintaining the cellular integrity. It is an inducible process that responds to the environment and it seems to be essential for cell survival during stress, starvation, hypoxia and consequently to the placenta implantation and development. Studies on the role of autophagy in placenta from pregnant women with preeclampsia are scarce and still unclear. The aim of this study was to compare the expression of autophagy-related proteins, LC3-II, beclin-1 as well as mTOR protein in placental tissue from pregnant women with preeclampsia and normotensive pregnant women. Methods: The expression of LC3-II and beclin-1 proteins involved in the autophagosome formation as well as mTOR protein was evaluated in 18 placentas of pregnant women with preeclampsia and in 20 of normotensive pregnant women who had elective cesarean delivery before the onset of labor. Fragments of the placenta were obtained immediately after delivery and the samples were prepared for real-time quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry and transmission electron microscopy analysis. Results: LC3-II expression as detected by RT-qPCR was significant higher in placentas of normotensive pregnant women compared with preeclamptic women. No significant difference in beclin-1 and mTOR expression was seen between these two groups. LC3-II, beclin-1 and mTOR proteins were detected by immunohistochemistry in syncytiotrophoblast and cytotrophoblast in both groups, and the presence of autophagic vacuoles containing intracytoplasmatic organelles was confirmed with electron microscopy. Conclusions: The results of the impaired autophagy demonstrated by decrease of LC3- II expression in placenta of
Abstracts / Pregnancy Hypertension: An International Journal of Women’s Cardiovascular Health 5 (2015) 2–52
preeclamptic patients might play a role in the pathophysiology of preeclampsia. Financial support: FAPESP 2013/00535-1 and 2012/ 24697-8. Disclosures: I.C. Weel: None. V.R. Ribeiro: None. M. Romao: None. M.L. Matias: None. V.T. Borges: None. J.C. Peracoli: None. J.P. Araujo Jr: None. M.T. Peracoli: None.
33
indices seem to be good ultrasonographic indicators for risk estimation and prevention of complications in pregnancies with hypertension. Disclosures: Á. Altorjay: None. A. Surányi: None. M. Jakó: None. T. Nyári: None. G. Németh: None. A. Pál: None. doi:10.1016/j.preghy.2014.10.067
doi:10.1016/j.preghy.2014.10.066
[64-OR] [63-OR] Effect of pregnancies complicated by essential as well as pregnancy induced hypertension on placental vascularization Ábel Altorjay a, Andrea Surányi a, Mária Jakó a, Tibor Nyári b, Gábor Németh a, Attila Pál a (a University of Szeged, Faculty of Medicine, Szeged, Hungary, b University of Szeged, Faculty of Physics and Informatics, Szeged, Hungary) Objectives: Proper uterine and placental vascularization is crucial for the adequate development of pregnancies. Pathological fetoplacental circulation leads to elevated resistance in placental vascularization which causes insufficiency and decreased fetal oxygenation. The aim of the study was to analyze placental vascularization in pregnant women with essential hypertension (EH) and pregnancy induced hypertension (PIH) with the help of VOCAL technique. Methods: We measured placental 3-dimensional power Doppler indices such as vascularization index (VI), flow index (FI) and vascularization flow index (VFI) in pregnancies complicated by PIH (NPIH = 40), and EH (NEH = 40) and compared these to pregnancies with normal blood pressure (NNBP = 316). We analyzed the correlation between the above mentioned indices, the volume of the placenta and placental localization. We used two-sample Wilcoxon rank-sum test, Levene’s test and quantile regression. Results: VI and VFI were significantly lower in pregnancies with PIH (VImean ± SD: 10.60 ± 10.40; VFImean ± SD: 2.77 ± 2.13) than in pregnancies with NBP (p < 0.001) and they were also significantly lower in case of PIH compared to EH (p < 0.01). We haven’t found significant difference between EH (VImean ± SD: 15.46 ± 8.49; VFImean ± SD: 7.55 ± 5.06) and NBP groups. FI was significantly lower in pregnancies with EH and PIH than in pregnancies with NBP (p < 0.001), and we found significant difference between EH (FImean ±SD: 43.89 ± 9.99) and PIH (FImean ±SD: 36.42 ± 13.67) as well. The placental volume was significantly smaller (zscore: 1.5) in EH as well as PIH than in NBP pregnancies, but there was no significant difference (zscore: 0.12) between the two affected groups. The placental localization had no significant influence on VI, FI or VFI (p < 0.01). There was no correlation found between the indices and gestational weeks. Conclusions: The direct 3-dimensional measurement of placental vascularization should gather more and more ground, because anomalies of uteroplacental and fetal vascularization appear too late, when complications have already been developed. Lower placental vascularization
Molecular basis for racial disparity in pre-eclampsia Elena R. Rivers a, Anthony J. Horton a, Christopher D. Clark a, Elizabeth G. Favre a, Katherine M. Senf a, Angela F. Hawk a, Eugene Y. Chang a, Christopher J. Robinson b, Kyu-Ho Lee a (a Medical University of South Carolina, b Charleston, SC, USA, University of Virginia, Charlottesville, VA, USA) Objectives: Pre-eclampsia (PE) affects 5-8% of pregnancies, and factors causing PE and mediating observed racial disparity are poorly understood. The cardiovascular gene Nkx2-5 is expressed in trophoblast cells and abnormal amnions are observed in mice Nkx2-5 mutant mice. We assessed Nkx25-related gene expression in placenta from normal and early onset and severe pre-eclampsia (EOSPE) pregnancies. Methods: IHC and qPCR assay of Nkx2.5 and target gene expression in normal term and EOSPE placenta. Results: Nkx2-5 is highly expressed in a subset of early EOSPE placentae, and is highly correlated with expression of the PE marker sFlt-1, and with an RNA splicing factor, Sam68. These correlations are significant in Caucasian, but not African American women. siRNA knockdown of Sam68 significantly impacts sFlt-1 expression in vitro, supporting a hypothesis that Nkx2-5 impacts EOSPE severity via upregulation of Sam68 and increased sFlt-1 expression. Other Nkx2-5 targets related to metabolic stress responses are also differentially activated in between Caucasians and African American women with EOSPE. Conclusions: Nkx2-5 may play a direct role in the genesis of PE via regulation of Sam68/sFlt-1, and serve as a common link to two other stress-related modifiers of EOSPE.