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Maternal and Placental Inflammation and Oxidative stress in Women of Advanced Maternal Age
Abstracts / Placenta 57 (2017) 225e335
pathological placental examination. Inter-observer agreement was in general good, but some microscopic criteria on e.g. maternal vascular malperfusion need to be specified. Conclusion: As long as there is no internationally valid, easily understandable, reproducible classification system on pathological placental diagnoses, national and international comparison of placental insufficiency with worst fetal outcome is hampered. Our classification system has contributed to the revision of diagnostic criteria for placental diagnosis in the Amsterdam placenta consensus group. http://dx.doi.org/10.1016/j.placenta.2017.07.029
S2.4. MATERNAL AND PLACENTAL INFLAMMATION AND OXIDATIVE STRESS IN WOMEN OF ADVANCED MATERNAL AGE Samantha Lean, Katie Stephens, Alexander Heazell, Rebecca Jones. Maternal and Fetal Health Research Centre, University of Manchester, Manchster, UK Introduction: Women of advanced maternal age (AMA, 35 years) are a high risk pregnancy cohort for complications associated with placental dysfunction, including stillbirth and fetal growth restriction. Placental dysfunction is evident in women of AMA, but the underlying mechanisms are unclear. We predict that markers of ageing (inflammation and oxidative stress) will be evident in serum and placentas of women of AMA, contributing to placental dysfunction. Methods: A prospective cohort study (MAMAS) collected maternal plasma and placental samples from mothers aged 20-30, 35-39 and 40 years. ELISA measured biomarkers of ageing and third trimester maternal blood (n¼40/group) and placentas post-delivery (n¼15/group). Comparisons were made between mothers with normal pregnancy outcome aged 20-30 vs. AMA (Kruskal Wallis), and AMA mothers with normal vs. composite adverse outcome (CAPO; Mann-Whitney). Results: Women of AMA with both normal and adverse outcomes had raised total antioxidant capacity (TAC) in maternal circulation and placenta (p<0.05). This was associated with a reduced level of lipid peroxidation in maternal serum and placenta from women with AMA with normal pregnancies (p<0.05), but not adverse outcomes. Women of AMA with normal outcomes had a pro-inflammatory bias, with reduced concentrations of anti-inflammatory cytokines, IL-10 and IL-1RA, in maternal serum and elevated placental pro-inflammatory IFN-ɣ, TNF-a, IL-8, IL-1a (p<0.050.01). These changes were also detected in pregnancies with adverse outcomes, with additional downregulation of placental anti-inflammatory cytokines (IL-4, IL-10, IL-1Ra; p<0.05). Discussion: These studies reveal age-related changes in systemic oxidative balance and inflammatory status in women of AMA. Elevated TAC may act to combat and prevent lipid peroxidation with normal outcomes. The proinflammatory status was more marked in the placenta and was exacerbated in those women of AMA with adverse outcomes. These data support the hypothesis that oxidative stress and inflammation may contribute to placental dysfunction in women of AMA. http://dx.doi.org/10.1016/j.placenta.2017.07.030
S2.5. CAN PLACENTAL HISTOPATHOLOGY LESIONS PREDICT RECURRENCE OF FETAL GROWTH RESTRICTION? Michal Levy 1, 2, Yossi Mizrachi 1, 2, Sophia Lytes 1, 2, Eran Weiner 1,2, Letizia Schreiber 1, 2, Michal Kovo 1,2. 1 Edith Wolfson Medical Center, Holon, Israel; 2 Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel Objective: To study the role of placental pathology in the prediction of recurrence of fetal growth restriction (FGR). Methods: The medical records and placental pathological reports of normotensive women who gave birth at 24-42 weeks, to neonates with
229
birthweight <10th % were reviewed. Patients were divided according to their subsequent pregnancy into those who developed or did not develop recurrent FGR. The clinical and pathological characteristics of the index pregnancies were compared between the groups. Prediction models were generated for FGR recurrence. Results: The recurrent FGR group (n¼70) was characterized by a higher rate of placental weight <10th percentile (p¼0.02), and higher neonatal to placental weight ratio (p¼0.003), as compared to the non-recurrent FGR group (n¼105). On multivariate logistic regression analysis placental maternal and fetal vascular malperfusion lesions and higher neonatal to placental weight ratio were all independently associated with recurrent FGR. Birth-weight <3rd % was the only clinical variable associated with recurrent FGR. A clinical-pathological prediction model was superior to a prediction model including only clinical variables (AUC 0.71 vs. 0.57). Conclusion: The presence of placental vascular malperfusion lesions and increased neonatal to placental weight ratio in the index pregnancy, are associated with recurrent FGR in subsequent pregnancy. http://dx.doi.org/10.1016/j.placenta.2017.07.031
S3.1. NEUTROPHIL INFILTRATION IN DECIDUA AND PREECLAMPSIA Caroline Dunk. Lunenfeld Tanenbaum Research Institute, Toronto, Canada Introduction: We previously identified a novel population of 2nd trimester tissue resident decidual neutrophils that possess a distinct pro-angiogenic phenotype compared to peripheral blood neutrophils (PBN). In TGFbdominant environments, ie. tumours N1 inflammatory neutrophils differentiate to a pro-angiogenic tumour promoting N2 phenotype. We hypothesized that the decidua similarly differentiates PBN into a N2-like phenotype that would support utero-placental development. As failure of uterine vascular transformation is associated with the development of preeclampsia we investigated the if the effect of decidua-secreted factors on N1/N2 and angiogenic gene expression was coprimised in PE women as compared to healthy women. Methods: PBN were isolated from healthy women at 16-18 and 26-29 weeks of gestation, and from preeclamptic women 26-35wk (PE) (n¼5-7 in each group). Isolated PBN were split into 2 treatment groups: 1) RPMI; and 2) 2nd trimester decidua-conditioned media (DCM). After 5h, RNA was isolated for RT-qPCR of angiogenic and N1/N2 factors. Neutrophil transendothelial invasion was assessed in the same groups. Finally placental bed sections from healthy non labouring women and severe early onset PE women were examined for the presence of infiltrated neutrophils using immunohistochemistry and image analysis. Results: Cluster analysis of 16 week, 26 week and PE samples identified segregating angiogenic and N1/N2 expression profiles of PBN from healthy and PE women under both SFM and DCM stimulated conditions. Interestingly one way ANOVA showed that in both analyses the PE samples grouped with the 16-18 week healthy samples and were distinctly different from their gestational matched 27-29 week controls. Levels of BAI1, CD59, RUNX1, TGFA, THBS1, FAS1, CXCR4 and TNF were increased in DCM stimulated PE PBN while CCL-2, CCL-5, CSF3, CXCL2 and 3, EREG, IL6, SERPINE1, CMYC, ICAM1 and VEGF were decreased. Three of these genes (CMYC, CXCR4, FAS) can reliably distinguish PE PBN from 26 week samples (binary logistic regression). In transendothelial invasion assays both 16-18 wk and 26-29wk samples migrated in response to both DCM and IL8. PE PBN did not migrate in response to IL8 and showed a blunted response to DCM. Finally while tissue infiltrated neutrophils are often observed in placental bed sample from healthy pregnancies they are absent from the PE placental bed. Conclusion: PE PBN resistance to decidual driven angioigenic N2 like differentiation may contribute to the inflammatory endothelial dysfunction and deficient uterovascular remodelling associated with PE. http://dx.doi.org/10.1016/j.placenta.2017.07.032
pathological placental examination. Inter-observer agreement was in general good, but some microscopic criteria on e.g. maternal vascular malperfusion need to be specified. Conclusion: As long as there is no internationally valid, easily understandable, reproducible classification system on pathological placental diagnoses, national and international comparison of placental insufficiency with worst fetal outcome is hampered. Our classification system has contributed to the revision of diagnostic criteria for placental diagnosis in the Amsterdam placenta consensus group. http://dx.doi.org/10.1016/j.placenta.2017.07.029
S2.4. MATERNAL AND PLACENTAL INFLAMMATION AND OXIDATIVE STRESS IN WOMEN OF ADVANCED MATERNAL AGE Samantha Lean, Katie Stephens, Alexander Heazell, Rebecca Jones. Maternal and Fetal Health Research Centre, University of Manchester, Manchster, UK Introduction: Women of advanced maternal age (AMA, 35 years) are a high risk pregnancy cohort for complications associated with placental dysfunction, including stillbirth and fetal growth restriction. Placental dysfunction is evident in women of AMA, but the underlying mechanisms are unclear. We predict that markers of ageing (inflammation and oxidative stress) will be evident in serum and placentas of women of AMA, contributing to placental dysfunction. Methods: A prospective cohort study (MAMAS) collected maternal plasma and placental samples from mothers aged 20-30, 35-39 and 40 years. ELISA measured biomarkers of ageing and third trimester maternal blood (n¼40/group) and placentas post-delivery (n¼15/group). Comparisons were made between mothers with normal pregnancy outcome aged 20-30 vs. AMA (Kruskal Wallis), and AMA mothers with normal vs. composite adverse outcome (CAPO; Mann-Whitney). Results: Women of AMA with both normal and adverse outcomes had raised total antioxidant capacity (TAC) in maternal circulation and placenta (p<0.05). This was associated with a reduced level of lipid peroxidation in maternal serum and placenta from women with AMA with normal pregnancies (p<0.05), but not adverse outcomes. Women of AMA with normal outcomes had a pro-inflammatory bias, with reduced concentrations of anti-inflammatory cytokines, IL-10 and IL-1RA, in maternal serum and elevated placental pro-inflammatory IFN-ɣ, TNF-a, IL-8, IL-1a (p<0.050.01). These changes were also detected in pregnancies with adverse outcomes, with additional downregulation of placental anti-inflammatory cytokines (IL-4, IL-10, IL-1Ra; p<0.05). Discussion: These studies reveal age-related changes in systemic oxidative balance and inflammatory status in women of AMA. Elevated TAC may act to combat and prevent lipid peroxidation with normal outcomes. The proinflammatory status was more marked in the placenta and was exacerbated in those women of AMA with adverse outcomes. These data support the hypothesis that oxidative stress and inflammation may contribute to placental dysfunction in women of AMA. http://dx.doi.org/10.1016/j.placenta.2017.07.030
S2.5. CAN PLACENTAL HISTOPATHOLOGY LESIONS PREDICT RECURRENCE OF FETAL GROWTH RESTRICTION? Michal Levy 1, 2, Yossi Mizrachi 1, 2, Sophia Lytes 1, 2, Eran Weiner 1,2, Letizia Schreiber 1, 2, Michal Kovo 1,2. 1 Edith Wolfson Medical Center, Holon, Israel; 2 Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel Objective: To study the role of placental pathology in the prediction of recurrence of fetal growth restriction (FGR). Methods: The medical records and placental pathological reports of normotensive women who gave birth at 24-42 weeks, to neonates with
229
birthweight <10th % were reviewed. Patients were divided according to their subsequent pregnancy into those who developed or did not develop recurrent FGR. The clinical and pathological characteristics of the index pregnancies were compared between the groups. Prediction models were generated for FGR recurrence. Results: The recurrent FGR group (n¼70) was characterized by a higher rate of placental weight <10th percentile (p¼0.02), and higher neonatal to placental weight ratio (p¼0.003), as compared to the non-recurrent FGR group (n¼105). On multivariate logistic regression analysis placental maternal and fetal vascular malperfusion lesions and higher neonatal to placental weight ratio were all independently associated with recurrent FGR. Birth-weight <3rd % was the only clinical variable associated with recurrent FGR. A clinical-pathological prediction model was superior to a prediction model including only clinical variables (AUC 0.71 vs. 0.57). Conclusion: The presence of placental vascular malperfusion lesions and increased neonatal to placental weight ratio in the index pregnancy, are associated with recurrent FGR in subsequent pregnancy. http://dx.doi.org/10.1016/j.placenta.2017.07.031
S3.1. NEUTROPHIL INFILTRATION IN DECIDUA AND PREECLAMPSIA Caroline Dunk. Lunenfeld Tanenbaum Research Institute, Toronto, Canada Introduction: We previously identified a novel population of 2nd trimester tissue resident decidual neutrophils that possess a distinct pro-angiogenic phenotype compared to peripheral blood neutrophils (PBN). In TGFbdominant environments, ie. tumours N1 inflammatory neutrophils differentiate to a pro-angiogenic tumour promoting N2 phenotype. We hypothesized that the decidua similarly differentiates PBN into a N2-like phenotype that would support utero-placental development. As failure of uterine vascular transformation is associated with the development of preeclampsia we investigated the if the effect of decidua-secreted factors on N1/N2 and angiogenic gene expression was coprimised in PE women as compared to healthy women. Methods: PBN were isolated from healthy women at 16-18 and 26-29 weeks of gestation, and from preeclamptic women 26-35wk (PE) (n¼5-7 in each group). Isolated PBN were split into 2 treatment groups: 1) RPMI; and 2) 2nd trimester decidua-conditioned media (DCM). After 5h, RNA was isolated for RT-qPCR of angiogenic and N1/N2 factors. Neutrophil transendothelial invasion was assessed in the same groups. Finally placental bed sections from healthy non labouring women and severe early onset PE women were examined for the presence of infiltrated neutrophils using immunohistochemistry and image analysis. Results: Cluster analysis of 16 week, 26 week and PE samples identified segregating angiogenic and N1/N2 expression profiles of PBN from healthy and PE women under both SFM and DCM stimulated conditions. Interestingly one way ANOVA showed that in both analyses the PE samples grouped with the 16-18 week healthy samples and were distinctly different from their gestational matched 27-29 week controls. Levels of BAI1, CD59, RUNX1, TGFA, THBS1, FAS1, CXCR4 and TNF were increased in DCM stimulated PE PBN while CCL-2, CCL-5, CSF3, CXCL2 and 3, EREG, IL6, SERPINE1, CMYC, ICAM1 and VEGF were decreased. Three of these genes (CMYC, CXCR4, FAS) can reliably distinguish PE PBN from 26 week samples (binary logistic regression). In transendothelial invasion assays both 16-18 wk and 26-29wk samples migrated in response to both DCM and IL8. PE PBN did not migrate in response to IL8 and showed a blunted response to DCM. Finally while tissue infiltrated neutrophils are often observed in placental bed sample from healthy pregnancies they are absent from the PE placental bed. Conclusion: PE PBN resistance to decidual driven angioigenic N2 like differentiation may contribute to the inflammatory endothelial dysfunction and deficient uterovascular remodelling associated with PE. http://dx.doi.org/10.1016/j.placenta.2017.07.032