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335: Gestational hypertension and induction of labor at 37 weeks: how changing guidelines have affected outcomes
Poster Session II during pregnancy. A prospective study was conducted on 44 pregnant women. During first half of pregnancy, noninvasive assessment of central aortic pressure and arterial stiffness was performed. The subjects were followed postpartum to determine maternal and fetal outcomes. We hypothesized that the non-invasive central blood pressure monitoring would identify pregnant women at higher risk for developing preeclampsia and GHTN during pregnancy. Differences in various indices of central blood pressure were measured. Independent students t test was used to compare mean differences in pulse pressure and augmentation 75 index between women with hypertensive disorders of pregnancy (preeclampsia alone, GHTN alone, and preeclampsia or GHTN combined) and normotensive women. RESULTS: Women with GHTN had a significantly higher mean pulse pressure than normotensive women (42.6 + 2.8 mm Hg versus 34.0 + 7.9 mm Hg, p<0.05). Women with either preeclampsia or GHTN had a significantly higher mean pulse pressure than normotensive women (42.7 + 9.5 mm Hg versus 34.0 + 7.9 mm Hg, p<0.05). Women with either preeclampsia or GHTN also had a significantly higher mean augmentation 75 index than normotensive women (30.7 + 20.1 versus 15.6 + 16.8, p<0.05) CONCLUSION: Women with hypertensive disorders of pregnancy have a significantly higher pulse pressure and augmentation index early in gestation compared to the normotensive women. Results suggest that noninvasive methods to measure arterial stiffness identify these patients earlier in pregnancy which can facilitate pregnancy management. Further studies are required to corroborate and extend these findings with larger sample sizes, particularly in women with pre eclampsia.
ajog.org 334 Regulation of CRLR and RAMPs by nitric oxide in endothelial cells Madhu Chauhan1, Meena Balakrishnan1, Chandra Yallampalli1 1
Baylor College of Medicine, Houston, TX
OBJECTIVE: Endothelium plays a major role in maintaining normal
blood pressure through several mechanisms and nitric oxide (NO) system is the primary one.NO is synthesized by NOS utilizing L-Arginine as a substrate. We have shown earlier that calcitonin gene related peptide (CGRP) family play a major role in vascular adaptation during pregnancy. A 7TM-calcitonin receptor like receptor (CRLR) and 3 receptor activity modifying proteins (RAMP1, 2, and 3) are the receptor components for CGRP family of peptides. However, it is unclear if NO regulates the levels of these receptors components in endothelium and thus the vascular adaptations in pregnancy. The aims of the study are to :1) Investigate the expression of CRLR and RAMPs in omental artery (OA) from pregnant normal (Preg) and PE women, 2) Assess the expression of CRLR and RAMPs in endothelial cells and, 3) Determine if NO donor DETA-NO and inhibitor L-NAME regulate receptor components levels in endothelial cells STUDY DESIGN: OA collected during caesarian section from women with Preg. (n¼5), and PE (n¼5) were used for RNA studies and immuno-fluorescent localization of receptor components. Uterine human microvascular endothelial cells (Ut-HMVEC) were treated with DETA-NO (10-6M-10-4M) or nitro-L-Arginine methyl ester (L-NAME) (10-6M-10-4M) for 24 hrs at 370C in a humidified incubator in an atmosphere of 95% air and 5% carbon dioxide. The mRNA levels were measured using qPCR and expressed relative to GAPDH. P<0.05 was considered significant. RESULTS: 1) Expression of mRNA levels of CRLR was higher (P< 0.01) in PE (2.28+/-0.56) compared to Preg (1.01+/-0.08) and no changes in RAMP 1 or 2 are noted. However, RAMP 3 mRNA levels are reduced in PE (0.18+/-0.08) compared to Preg (1.17+/-0.25, 2) RAMP 2 but not RAMP1 and RAMP3 mRNA was detected in the UtMVEC cells, 3) All 3 RAMPs are present in both, smooth muscle and endothelial cells in the OA, 4) DETA-NO caused a decline (P<0.05) whereas, inhibitor of NO synthesis, L-NAME caused an increase in the expression of CRLR and RAMP2 in Ut-HMVEC cells (P<0.05) CONCLUSION: Deficiency of NO production in vascular endothelial cells from PE women could trigger an increase in endothelial CRLR and RAMP2 and, thus the vasodialatory effects of CGRP family peptides as a compensatory mechanism to control blood pressure.
335 Gestational hypertension and induction of labor at 37 weeks: how changing guidelines have affected outcomes Katherine Betcher1 1
William Beaumont Hospital, Royal Oak, MI
OBJECTIVE: We sought to compare pregnancy outcomes for women
presenting for induction of labor with gestational hypertension or preeclampsia without severe features in the year preceding and following the publication of the “Task Force on Hypertension in Pregnancy” document by the American College of Obstetricians and Gynecologists. This document suggested delivery when diagnosis of hypertension without severe features was made if over 37 weeks gestational age. STUDY DESIGN: All women with indication for induction of either gestational hypertension or preeclampsia without severe features were reviewed over a 2 year period at a single institution (n¼269); 136 were induced prior to hypertension guideline changes, and 133 were induced after guideline changes. Evaluated outcomes include
S188 American Journal of Obstetrics & Gynecology Supplement to JANUARY 2016
Poster Session II
ajog.org primary cesarean rate, progression to severe disease, need for magnesium sulfate treatment, and need for anti-hypertensive medication. RESULTS: 79% (n¼212) of patients had gestational hypertension, 21% (n¼57) had pre-eclampsia without severe features. Average BMI was 35.1 in the pre-guideline changes group and 36.2 in the post- changes group (p¼0.24). Overall, 40.4% (n¼53) of patients in the pre-guideline changes group had a C-section, vs. 31.6% (n¼42) of patients in the post-guideline change group (p¼0.13). There were similar numbers of failed inductions in the pre- and post- guidelines groups: 9.4% (n¼5) and 14.6% (n¼6). More women in the preguidelines group required antihypertensives in labor; 26.1% vs 8.3% (p¼0.0001). More women in the pre-guidelines group developed severe symptoms during and after labor; 41.2% vs 21.8% (p¼0.0006), and more were administered magnesium therapy; 26.5% vs 9.0% (p¼0.0002). CONCLUSION: Practice guidelines changed to recommend induction of labor for hypertensive disease at an earlier gestational age. The study group post-guidelines trended toward lower rates of cesarean delivery and had significantly less progression to severe disease. These findings support the original HYPITAT study group data in an American population with high rates of obesity.
336 Why is the placenta not a fetus? Identifying silenced gene targets in the placenta at a transcriptome-wide scale using AGO-CLIP sequencing technology Kjersti Aagaard1, Mark Hamilton1, Maxim Seferovic1, Diana Racusin1, Sean McGuire1,2 1
Baylor College of Medicine, Houston, TX, 2University of Texas MD Anderson Cancer Center, Houston, TX
OBJECTIVE: Eutherian mammals, and humans in particular, rest on a fundamental biologic paradox. Soon after conception, the blastocyst exquisitely delineates into two lineages of identical genetic complement: the trophoblast (placenta) and the inner cell mass (embryo). Because prevalent disorders, such as preeclampsia, are intimately tied trophoblast differentiation, understanding how the fetal transcriptome is silenced while the trophoblast (placental) transcriptome is activated is essential. Based on established evidence that placentaspecific miRNAs (such as C19MC) serve as potent gene silencers by binding the 3’ untranslated region (3’UTR), we hypothesized that thorough identification of placental miRNA targets at a transcriptome wide scale would lead to breakthroughs in our understanding of placental biology and pathology. STUDY DESIGN: Argonaute protein cross-linking immunoprecipitation coupled to high-throughput sequencing (AGO-CLIP-seq) directly defines the discoverable miRNA interactome. AGO proteins serve as the key effector molecules that bind to miRNAs and mediate target recognition, canonically in the mRNA 3’UTR. Using the Illumina TruSeq small RNA platform we performed AGO-CLIP-seq in healthy, preterm, and preeclamptic primary human placental tissue (n¼9). Analytics were on customized mapping pipelines, and annotated against human assemblies (Gencode). RESULTS: The results of the AGO-CLIP-seq analyses are shown (Figures). Consistent with placental-specific patterning, AGOmiRNA loading demonstrates a clear predominance (w28% of total expression) of C19MC placental-specific miRNA coverage. C19MCcontaining miRNA family targets are also significantly enriched in the 3’UTR accounting for 25% (115/447) of unique targets (Fig. 1). Among preeclamptic samples (n¼4), there was significant depletion of mitochondrial gene targets and their cognate pathways (Fig. 2). CONCLUSION: This study is the first direct genome-wide identification of miRNA targets in the human placenta. Collectively, our findings identify multiple unique C19MC miRNA targets in normal & preeclamptic subjects. We speculate that miRNA mediated RNA control is a fundamental & rapid molecular means of regulating the transcriptome in placental differentiation.
Supplement to JANUARY 2016 American Journal of Obstetrics & Gynecology
S189
ajog.org 334 Regulation of CRLR and RAMPs by nitric oxide in endothelial cells Madhu Chauhan1, Meena Balakrishnan1, Chandra Yallampalli1 1
Baylor College of Medicine, Houston, TX
OBJECTIVE: Endothelium plays a major role in maintaining normal
blood pressure through several mechanisms and nitric oxide (NO) system is the primary one.NO is synthesized by NOS utilizing L-Arginine as a substrate. We have shown earlier that calcitonin gene related peptide (CGRP) family play a major role in vascular adaptation during pregnancy. A 7TM-calcitonin receptor like receptor (CRLR) and 3 receptor activity modifying proteins (RAMP1, 2, and 3) are the receptor components for CGRP family of peptides. However, it is unclear if NO regulates the levels of these receptors components in endothelium and thus the vascular adaptations in pregnancy. The aims of the study are to :1) Investigate the expression of CRLR and RAMPs in omental artery (OA) from pregnant normal (Preg) and PE women, 2) Assess the expression of CRLR and RAMPs in endothelial cells and, 3) Determine if NO donor DETA-NO and inhibitor L-NAME regulate receptor components levels in endothelial cells STUDY DESIGN: OA collected during caesarian section from women with Preg. (n¼5), and PE (n¼5) were used for RNA studies and immuno-fluorescent localization of receptor components. Uterine human microvascular endothelial cells (Ut-HMVEC) were treated with DETA-NO (10-6M-10-4M) or nitro-L-Arginine methyl ester (L-NAME) (10-6M-10-4M) for 24 hrs at 370C in a humidified incubator in an atmosphere of 95% air and 5% carbon dioxide. The mRNA levels were measured using qPCR and expressed relative to GAPDH. P<0.05 was considered significant. RESULTS: 1) Expression of mRNA levels of CRLR was higher (P< 0.01) in PE (2.28+/-0.56) compared to Preg (1.01+/-0.08) and no changes in RAMP 1 or 2 are noted. However, RAMP 3 mRNA levels are reduced in PE (0.18+/-0.08) compared to Preg (1.17+/-0.25, 2) RAMP 2 but not RAMP1 and RAMP3 mRNA was detected in the UtMVEC cells, 3) All 3 RAMPs are present in both, smooth muscle and endothelial cells in the OA, 4) DETA-NO caused a decline (P<0.05) whereas, inhibitor of NO synthesis, L-NAME caused an increase in the expression of CRLR and RAMP2 in Ut-HMVEC cells (P<0.05) CONCLUSION: Deficiency of NO production in vascular endothelial cells from PE women could trigger an increase in endothelial CRLR and RAMP2 and, thus the vasodialatory effects of CGRP family peptides as a compensatory mechanism to control blood pressure.
335 Gestational hypertension and induction of labor at 37 weeks: how changing guidelines have affected outcomes Katherine Betcher1 1
William Beaumont Hospital, Royal Oak, MI
OBJECTIVE: We sought to compare pregnancy outcomes for women
presenting for induction of labor with gestational hypertension or preeclampsia without severe features in the year preceding and following the publication of the “Task Force on Hypertension in Pregnancy” document by the American College of Obstetricians and Gynecologists. This document suggested delivery when diagnosis of hypertension without severe features was made if over 37 weeks gestational age. STUDY DESIGN: All women with indication for induction of either gestational hypertension or preeclampsia without severe features were reviewed over a 2 year period at a single institution (n¼269); 136 were induced prior to hypertension guideline changes, and 133 were induced after guideline changes. Evaluated outcomes include
S188 American Journal of Obstetrics & Gynecology Supplement to JANUARY 2016
Poster Session II
ajog.org primary cesarean rate, progression to severe disease, need for magnesium sulfate treatment, and need for anti-hypertensive medication. RESULTS: 79% (n¼212) of patients had gestational hypertension, 21% (n¼57) had pre-eclampsia without severe features. Average BMI was 35.1 in the pre-guideline changes group and 36.2 in the post- changes group (p¼0.24). Overall, 40.4% (n¼53) of patients in the pre-guideline changes group had a C-section, vs. 31.6% (n¼42) of patients in the post-guideline change group (p¼0.13). There were similar numbers of failed inductions in the pre- and post- guidelines groups: 9.4% (n¼5) and 14.6% (n¼6). More women in the preguidelines group required antihypertensives in labor; 26.1% vs 8.3% (p¼0.0001). More women in the pre-guidelines group developed severe symptoms during and after labor; 41.2% vs 21.8% (p¼0.0006), and more were administered magnesium therapy; 26.5% vs 9.0% (p¼0.0002). CONCLUSION: Practice guidelines changed to recommend induction of labor for hypertensive disease at an earlier gestational age. The study group post-guidelines trended toward lower rates of cesarean delivery and had significantly less progression to severe disease. These findings support the original HYPITAT study group data in an American population with high rates of obesity.
336 Why is the placenta not a fetus? Identifying silenced gene targets in the placenta at a transcriptome-wide scale using AGO-CLIP sequencing technology Kjersti Aagaard1, Mark Hamilton1, Maxim Seferovic1, Diana Racusin1, Sean McGuire1,2 1
Baylor College of Medicine, Houston, TX, 2University of Texas MD Anderson Cancer Center, Houston, TX
OBJECTIVE: Eutherian mammals, and humans in particular, rest on a fundamental biologic paradox. Soon after conception, the blastocyst exquisitely delineates into two lineages of identical genetic complement: the trophoblast (placenta) and the inner cell mass (embryo). Because prevalent disorders, such as preeclampsia, are intimately tied trophoblast differentiation, understanding how the fetal transcriptome is silenced while the trophoblast (placental) transcriptome is activated is essential. Based on established evidence that placentaspecific miRNAs (such as C19MC) serve as potent gene silencers by binding the 3’ untranslated region (3’UTR), we hypothesized that thorough identification of placental miRNA targets at a transcriptome wide scale would lead to breakthroughs in our understanding of placental biology and pathology. STUDY DESIGN: Argonaute protein cross-linking immunoprecipitation coupled to high-throughput sequencing (AGO-CLIP-seq) directly defines the discoverable miRNA interactome. AGO proteins serve as the key effector molecules that bind to miRNAs and mediate target recognition, canonically in the mRNA 3’UTR. Using the Illumina TruSeq small RNA platform we performed AGO-CLIP-seq in healthy, preterm, and preeclamptic primary human placental tissue (n¼9). Analytics were on customized mapping pipelines, and annotated against human assemblies (Gencode). RESULTS: The results of the AGO-CLIP-seq analyses are shown (Figures). Consistent with placental-specific patterning, AGOmiRNA loading demonstrates a clear predominance (w28% of total expression) of C19MC placental-specific miRNA coverage. C19MCcontaining miRNA family targets are also significantly enriched in the 3’UTR accounting for 25% (115/447) of unique targets (Fig. 1). Among preeclamptic samples (n¼4), there was significant depletion of mitochondrial gene targets and their cognate pathways (Fig. 2). CONCLUSION: This study is the first direct genome-wide identification of miRNA targets in the human placenta. Collectively, our findings identify multiple unique C19MC miRNA targets in normal & preeclamptic subjects. We speculate that miRNA mediated RNA control is a fundamental & rapid molecular means of regulating the transcriptome in placental differentiation.
Supplement to JANUARY 2016 American Journal of Obstetrics & Gynecology
S189