Enhanced expression of the stemness-related factors OCT4, SOX15 and TWIST1 in ectopic endometrium of endometriosis patients

Abstract Background Current evidence suggests that endometrial-derived stem cells, spilled in the peritoneal cavity via retrograde menstruation, are key players in the establishment of endometriotic lesions. The aim of this study was to determine the presence and distribution of the stemness-related factors OCT4, SOX15, TWIST1 and DCAMLK1 in women with and without endometriosis. Methods Immunohistochemical analysis was used to determine stromal and epithelial expression of OCT4, SOX15, TWIST1 and DCAMLK1 in endometriosis patient (EP) endometrium (n = 69) and endometriotic tissue (n = 90) and in control endometrium (n = 50). Quantitative Real-Time PCR of OCT4, SOX15 TWIST1 and DCAMLK1 was performed in paired samples of EP endometrium and endometriotic tissue. Co-immunofluorescence staining was performed for OCT4 and SOX15. For statistical analyses we used unpaired t-test, Fisher combination test and Spearman test. For paired analyses, paired t-test and McNemar test were used. Results We detected a significant correlation between the expression of the established stem cell marker OCT4 and the stemness-related markers SOX15 (p < 0.001) and TWIST1 (p = 0.002) but not DCAMLK1. We showed a colocalization of SOX15 and OCT4 in epithelial and stromal cells of endometriotic tissue by coimmunofluorescence. A concordant expression of OCT4 and SOX15 in the same sample was observed in epithelial cells of the endometriotic tissue (71.7%). The expression of stemness-related factors was not associated with proliferative or secretory phase of the menstrual cycle in endometriosis patients but was found to be differentially expressed during the menstrual cycle in the control group. Increased expression of epithelial OCT4, SOX15 and TWIST1 was detected in endometriotic tissue compared to EP endometrium in paired (p = 0.021, p < 0.001 and p < 0.001) and unpaired analysis (p = 0.040, p < 0.001 and p = 0.001). Conclusion Our findings support the hypothesis that upregulation of stem cell-related factors contribute to the establishment of endometriotic lesions. Trial registration The study was approved by the institutional review board (545/2010 on 6th of May 2014) of the Medical University of Vienna ( http://ethikkommission.meduniwien.ac.at/fileadmin/ethik/media/dokumente/register/alle_2010.pdf )

Expression of angiogenetic factors in the eutopic endometrium in women with and without endometriosis and ectopic lesions

Unter Endometriose versteht man das Vorkommen endometriumhaltiger Zellen außerhalb des Cavum uteri. Der genaue Entstehungsmechanismus ist noch nicht geklärt. Man geht von einem multifaktoriellen Geschehen aus, bei der die Angiogenese eine große Rolle spielt. Das Ziel dieser Arbeit war es, das Expressionsverhalten verschiedener Angiogenesefaktoren zu beschreiben. Folgende Faktoren wurden untersucht: VEGFA, VEGFR2, HIF1A,HGF,PDGFB, FGF 18, TNFa, TGFB2, EPO, ANG,EPHB4,NRP1.Ihr Expressionsmuster wurde bei gesunden und erkrankten Frauen, im eutopen Endometrium und im ektopem Gewebe verglichen. Ferner wurden unterschiedliche Gewebsproben der einen und derselben Patientin sowie das Expressionsmuster in unterschiedlichen ektopen Läsionen (Ovar, Peritoneum, Rektum) untersucht. Insgesamt wurden 205 Gewebsproben von 114 Frauen aufgearbeitet. Annährend alle Angiogenesefaktoren zeigten nennenswerte Anstiege im eutopen Endometrium bei Frauen mit Endometriose und 6 Faktoren (VEGFR2, HIF1A, HGF, PDGFB, NRP1 und EPHB4) im ektopen Läsionen im Vergleich zum Endometrium gesunder und kranker Frauen. Eine Hochregulation von VEGFR2, HIF1A und EPHB4 in eutopen Läsion konnte festgestellt werden. Für HIF1A, HGF und EPHB4 konnte keine significante hocregulation im Peritonealgewebe gezeigt werden, welches nicht direct von Endometriose betroffen war. Eine starke Korrellation wurde für drei Gene gefunden VEGFR2, PDGFB und EPHB4. Die Ergebnisse lassen den Schluss zu, dass unterschiedliche Angiogenesefaktoren bei der Entstehung der Endometriose beteiligt sind. Diese Faktoren sind unterschiedlich stark am Geschehen beteiligt und voneinander abhängig.Endometriosis is one of the most common benign gynecological diseases in women of reproductive age. Endometriosis is a multifactorial event, in which angiogenesis plays a major role. Angiogenesis is a prerequisite for the formation of endometriotic lesions. In fact, the survival of endometriotic lesions depends on adequate blood supply. A number of angiogenetic factors have been investigated and reported on in literature, but there is little information about the interaction of these factors.The objective of the present study was to evaluate the expression of vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor receptor 2 (VEGFR2), hypoxia-inducible factor (HIF1A), hepatocyte growth factor (HGF), platelet-derived growth factor (PDGFB), fibroblast growth factor 18 (FGF 18), tumor necrosis factor alpha (TNFa), transforming growth factor beta 2; (TGFB2), erythropoietin (EPO), angiogenin (ANG), ephrin receptor EphB4 (EPHB4), and neuropilin 1(NRP1), in the eutopic endometrium of women with and without endometriosis and ectopic lesions (in paired samples). The expression pattern of these factors with reference to the location of the ectopic lesion and gene expression in the peritoneum, which is not directly affected by endometriosis, were also investigated.Potential differences in the expression pattern with reference to cycle phases were studied, hormone therapies determined, and rAFS staging was performed. We also looked for a potential interrelationship between these factors, particularly with regard to the impact of VEGF. 205 tissue samples of 114 women were studied: 61 women with endometriosis and 53 without.151 endometriotic tissue samples, (including those from the eutopic and ectopic endometrium) and 54 peritoneal tissue samples were investigated. The samples were obtained during an operative laparoscopy. For detection and quantification of mRNA expression, each sample was analyzed by quantitative real-time PCR after RNA extraction. All samples revealed marked gene expression in the eutopic endometrium of women with endometriosis and ectopic lesions, in contrast to controls. No expression was registered for EPO or ANG in any of the three groups, and therefore all further investigation in regard of these entities was omitted. TNF[alpha] gene expression revealed no significant correlation in terms of tissue type or lesion location. We demonstrated that VEGFR2, HIF1A, HGF, PDGFB, NRP1 and EPHB4 were overexpressed in ectopic lesions compared to eutopic tissues of women with endometriosis. We further showed a significant upregulation of VEGFR2, HIF1A and EPHB4 in eutopic endometrial tissues of women with endometriosis compared to that of controls. Additionally, a significant downregulation for HIF1A, HGF and EPHB4 was observed in unaffected peritoneal tissues of women with endometriosis compared to controls. Interestingly, HIF1A, EPHB4 and HGF show an inverse expression in peritoneal tissues compared to eutopic endometrial tissues. Further, strong correlations were found between the three genes VEGFR2, PDGFB, and EPHB4.In light of the fact that not only VEGFA but also VEGFR2, HIF1A, PDGFB, NRP1 and EBHB4 are highly expressed in endometriotic implants, it is reasonable to speculate that beside VEGFA other angiogenic factors may play a crucial role in the progression of endometriosis or rather in angiogenesis and vascularization of endometrial implants. Our expression analysis of various angiogenetic factors has highlighted some new putative candidates regarding an association with endometriosis (EPHB4, PDGFB) and has confrimed already known candidates (VEGFA, VEGFR2, and HIF1A). Further studies should be performed to discover the exact role of these genes in the pathogenesis of endometriosis. ^In addition, a higher focus upon non-lesional, unaffected peritoneum of endometriosis patients seems beneficially in the context of endometriosis.submitted by Gülen YerlikayaArbeit an der Bibliothek noch nicht eingelangt - Daten nicht geprüftAbweichender Titel laut Übersetzung der Verfasserin/des VerfassersMedizinische Universität Wien, Dissertation, 2015OeBB(VLID)271540

Archives of Gynecology and Obstetrics / Prenatal detection of right aortic arch

Purpose To examine an unselective population of fetuses with right aortic arch (RAA) and suggest perinatal management. Second, to evaluate the importance and possible implication of fetal MRI in those cases. Methods Retrospective study of 36 patients with RAA diagnosed prenatally between 2006 and 2017 in a tertiary referral center. Results 32 fetuses were diagnosed with RAA and 4 with double aortic arch (DAA). 7 (19, 5%) cases had intracardiac abnormalities. Tetralogy of the Fallot was the most frequent one. Other extracardiac malformations were observed in 11/36 (30, 6%). Karyotype was available in 16 (44, 5%) cases. Two had 22q11.2 microdeletion, two trisomy 21, and one 20p12.2 duplication. Two needed surgery for respiratory symptoms. A newborn was identified with epilepsy, LennoxGastaud syndrome and PallisterKillian syndrome postnatally and another one with showed hyperreflexia and premature closer of the fontanelle. Three feticides were performed for pregnancy termination in one case with 22q11 deletion, trisomy 21, and partial agenesis of corpus callosum. Conclusion RAA can be detected by fetal echocardiography and it is associated with other cardiac or non-cardiac abnormalities, 22q11 microdeletion, trisomy 21, and other chromosomal abnormalities. karyotyping should be offered in any case of RAA, irrespective of co-existing malformations. Fetal MRI is a promising diagnostic tool for assessment of congenital cardiovascular abnormalities and extracardiac anomalies.(VLID)496249

Simpson-Golabi-Behmel-Syndrome in Dichorionic-Diamniotic Twin Pregnancy

Simpson-Golabi-Behmel syndrome (SGBS) is a rare x-linked overgrowth syndrome with distinct clinical features, which is difficult to diagnose prenatally. We report the diagnosis of SGBS in dichorionic-diamniotic twin pregnancies in the first trimester by ultrasound and genetic testing. The affected fetus developed polyhydramnios and the cervical length of the mother decreased significantly. To save the unaffected twin, a selective feticide of the affected fetus was performed. Finally, the patient underwent preterm caesarean section due to premature rupture of membranes in the dead twin, and also intrauterine infection. While SGBS has been reported, this was the first case in a multiple pregnancy, with possible consequences for the healthy twin. In conclusion, SGBS is a rare condition, which should be considered in the differential diagnosis of prenatal overgrowth syndromes and associated malformation

Factors Influencing the Duration of Termination of Pregnancy for Fetal Anomaly with Mifepristone in Combination with Misoprostol

This study&rsquo;s aim was to determine relevant factors that influence the time interval between first induction and fetal expulsion in late termination of pregnancy (TOP) and TOP after previous feticide for severe fetal malformation with a mifepristone&ndash;misoprostol regime. This retrospective study included 913 TOPs from a single tertiary care referral center. In 197 out of 913 TOPs, a previous feticide had been performed due to advanced gestational age (after 22 + 0 weeks of gestation). Induction was accomplished using 600 mg mifepristone followed by 400 &mu;g misoprostol. The interval between first induction with misoprostol and fetal expulsion was examined. Univariate and multivariate logistic regression analysis were used to predict an induction interval of 12 h or less. The median gestational age at induction of labor was 18.9 weeks of pregnancy. In 487 (53.3%) cases women delivered within 12 h; in 344 (37.7%) cases the induction interval was between 12 h and 36 h. In 82 (9%) cases induction took longer than 36 h. Factors that were significantly associated with a delivery duration of &lt;12 h were a lower gestational age at induction (OR 0.87; 95% CI 0.84&ndash;0.89; p &lt; 0.001) and a history of at least one previous vaginal delivery (OR 1.57; 95% CI 1.20&ndash;2.05; p &lt; 0.001). Factors that had no impact included previous cesarean section, performing feticide before induction and maternal age. Maternal BMI showed a non-significant trend

Clinical Course and Outcome of Non-Immune Fetal Hydrops in Singleton Pregnancies

Nonimmune fetal hydrops is a condition defined by abnormal fluid accumulation in two or more body compartments. The aim is to evaluate factors associated with adverse outcome in diagnosed fetal hydrops and to investigate the aspects for the decision making in the case of termination of pregnancy. Therefore, a retrospective data analysis of pregnancies complicated by non-immune hydrops fetalis between 2004 and 2018 was performed in a single tertiary referral center. Of 361 pregnancies with diagnosed fetal hydrops, in 183 cases (50.7%), the parents decided to terminate the pregnancy. A strong relationship between etiology and termination of pregnancy was demonstrated, whereas the highest rates of termination of pregnancy were found if a chromosomal aberration was diagnosed. Of the remaining 178 cases, 51 cases (28.7%) had a miscarriage, 33 cases (18.5%) had an intrauterine fetal death, and 94 cases (52.8%) were live born, whereas 26 (27.7%) of these offspring died within the first week of life. The risk of an adverse outcome increased with lower gestational age at diagnosis (p &lt; 0.001). A nuchal translucency thickness greater than 2.5 mm was associated with an adverse outcome (p &lt; 0.01). Furthermore, pregnancies with adverse outcome had significantly more affected compartments (median: 3; IQR 2), compared with live born cases (median: 2; IQR 1; p &lt; 0.01). In conclusion, adverse outcome in pregnancies with fetal hydrops was associated with a lower gestational age at diagnosis, nuchal translucency greater than 2.5 mm and a higher count of affected compartments. These results confirm that a precise clinical workup to identify the underlying etiology of non-immune fetal hydrops is essential for a better prognostic assessment and accurate counselling of parents

To Predict the Requirement of Pharmacotherapy by OGTT Glucose Levels in Women with GDM Classified by the IADPSG Criteria

The aim of this study was to assess the association between OGTT glucose levels and requirement of pharmacotherapy in GDM patients classified by the IADPSG criteria. This study included 203 GDM patients (108 managed with lifestyle modification and 95 requiring pharmacotherapy). Clinical risk factors and OGTT glucose concentrations at 0 (G0), 60 (G60), and 120 min (G120) were collected. OGTT glucose levels were significantly associated with the later requirement of pharmacotherapy (ROC-AUC: 71.1, 95% CI: 63.8–78.3). Also, the combination of clinical risk factors (age, BMI, parity, and pharmacotherapy in previous gestation) showed an acceptable predictive accuracy (ROC-AUC: 72.1, 95% CI: 65.0–79.2), which was further improved when glycemic parameters were added (ROC-AUC: 77.5, 95% CI: 71.5–83.9). Random forest analysis revealed the highest variable importance for G0, G60, and age. OGTT glucose measures in addition to clinical risk factors showed promising properties for risk stratification in GDM patients classified by the recently established IADPSG criteria

Fetal Growth and Adipose Fat Tissue Trajectories in Twin Pregnancies after Gastric Bypass Surgery

Introduction!#!Previous studies demonstrated a continuous decline in fetal growth throughout singleton pregnancy after bariatric surgery. However, intrauterine growth in twin pregnancy is subjected to further underlying processes. This study was to investigate the longitudinal assessment of fetal biometry and abdominal fat thickness of twin pregnancies conceived after gastric bypass (GB) surgery and compare them to body mass index-matched (BMIM) and obese (OB) controls.!##!Materials and methods!#!We retrospectively assessed ultrasound data of 30 women with dichorionic-diamniotic twin pregnancy (11 women after GB surgery, 9 OB mothers with pregestational BMI ≥30 kg/m2, and 10 BMIM and age-matched controls). We assessed fetal growth parameters including fetal subcutaneous adipose tissue thickness (FSCTT) as well as newborn biometry after delivery. Patient characteristics were obtained from the medical records.!##!Results!#!The rise in FSCTT curves was markedly slower in the twin offspring of women with history of GB as compared to the offspring of OB mothers and offspring of BMIM controls. Hence, FSCTT was significantly decreased in the GB offspring as compared to both control groups at 34 weeks of gestation. Also, growth curves of abdominal circumference were decreased in the offspring of GB patients as compared to OB mothers. Infants of mothers with history of GB showed significantly lower birth weight percentiles compared to newborns of OB mothers (27.2 vs. 48.8 pct, p = 0.025). There was no significant difference in inter-twin birth weight difference between the offspring of GB (median: 9.9%, interquartile ranges [IQR]: 6.5-20.0) versus OB (median: 14.6%, IQR: 8.2-21.6) and BMIM controls (median: 9.0%, IQR: 6.3-12.6, p = 0.714).!##!Conclusions!#!In summary, intrauterine growth delay in twin pregnancies after GB is assumed to be a multifactorial event with altered metabolism as the most important factor. However, special attention must be paid to the particularity of twin pregnancies as they seem to be subject to other additional mechanism

Enhanced epithelial to mesenchymal transition (EMT) and upregulated MYC in ectopic lesions contribute independently to endometriosis

Abstract Background Epithelial to mesenchymal transition (EMT) is a process in which epithelial cells lose polarity and cell-to-cell contacts and acquire the migratory and invasive abilities of mesenchymal cells. These abilities are thought to be prerequisites for the establishment of endometriotic lesions. A hallmark of EMT is the functional loss of E-cadherin (CDH1) expression in epithelial cells. TWIST1, a transcription factor that represses E-cadherin transcription, is among the EMT inducers. SNAIL, a zinc-finger transcription factor, and its close relative SLUG have similar properties to TWIST1 and are thus also EMT inducers. MYC, which is upregulated by estrogens in the uterus by an estrogen response cis-acting element (ERE) in its promoter, is associated with proliferation in endometriosis. The role of EMT and proliferation in the pathogenesis of endometriosis was evaluated by analyzing TWIST1, CDH1 and MYC expression. Methods CDH1, TWIST1, SNAIL and SLUG mRNA expression was analyzed by qRT-PCR from 47 controls and 74 patients with endometriosis. Approximately 42 ectopic and 62 eutopic endometrial tissues, of which 30 were matched samples, were collected during the same surgical procedure. We evaluated TWIST1 and MYC protein expression by immunohistochemistry (IHC) in the epithelial and stromal tissue of 69 eutopic and 90 ectopic endometrium samples, of which 49 matched samples were analyzed from the same patient. Concordant expression of TWIST1/SNAIL/SLUG and CDH1 but also of TWIST1 and MYC was analyzed. Results We found that TWIST1, SNAIL and SLUG are overexpressed (p < 0.001, p = 0.016 and p < 0.001) in endometriosis, while CDH1 expression was concordantly reduced in these samples (p < 0.001). Similar to TWIST1, the epithelial expression of MYC was also significantly enhanced in ectopic endometrium compared to eutopic tissues (p = 0.008). We found exclusive expression of either TWIST1 or MYC in the same samples (p = 0.003). Conclusions Epithelial TWIST1 is overexpressed in endometriosis and may contribute to the formation of endometriotic lesions by inducing epithelial to mesenchymal transition, as CDH1 was reduced in ectopic lesions. We found exclusive expression of either TWIST1 or MYC in the same samples, indicating that EMT and proliferation contribute independently of each other to the formation of endometriotic lesions