Ambros Huber

Ten estrogen- related polymorphisms and endometriosis : A study of multiple gene-gene interactions

OBJECTIVE: Genetic as well as hormonal factors are known to influence the development and clinical course of endometriosis. We aimed to investigate the association among 10 single nucleotide polymorphisms (SNPs) involved in the estrogen metabolism and endometriosis and to develop a multiple genetic model. METHODS: In a case–control study, we investigated the genotype frequencies of 10 estrogen metabolizing SNPs in 32 patients with endometriosis and 790 healthy controls using sequencing-on-chip-technology with solid-phase polymerase chain reaction on oligonucleotide microarrays: catechol-O-methyltransferase, Val158Met G->A, 17-β-hydroxysteroid dehydrogenase type 1 (HSD17), vlV A->C, cytochrome P450 (CYP), 17 A2 allele T->C, CYP1A1 MspI RFLP T->C, CYP1A1 Ile462Val A->G, CYP19 Arg264Cys C->T, CYP19 C1558T C->T, CYP 1B1 Leu432Val, CYP1B1 Asn453Ser, and estrogen receptor alpha IVS1 –401>C. Associations and 2-way interaction models between SNPs were calculated by stepwise logistic regression models. RESULTS: In a univariate model, HSD17 vlV A->C was associated with a significantly increased risk of endometriosis (P = .004; odds ratio 3.9, 95% confidence interval 1.6–9.8). When all 2-way interactions of investigated SNPs were ascertained, no significant interactions among SNPs were observed. In a multivariate model, HSD17 vlV A->C was also significantly associated with endometriosis (P = .002). CONCLUSION: We present data on multiple SNPs in patients with endometriosis indicating an association between HSD17 gene variation and the disease. Although not able to demonstrate interaction models of SNPs, we provide evidence of HSD17 vlV A->C as a low penetrance genetic marker of endometriosis. LEVEL OF EVIDENCE: II-2

Prevalence of Cervical and Intrauterine Human Papillomavirus Infected in the Third Trimester in Asymptomatic Women

Objective: To study the prevalence and association of human papillomavirus (HPV) infection in the cervix of pregnant women without visible signs of genital HPV infection undergoing cesarean delivery in the third trimester and to investigate a possible HPV transmission to the fetus. Methods: All women underwent cesarean delivery between 37 and 40 weeks of gestation. Cervical samples were taken prior to cesarean delivery. Furthermore, amniotic fluid, placental tissue, and cord blood were sampled and polymerase chain reaction (PCR) or Hybrid Capture II test (Digene Corp, Beltsville, MD) was performed to detect HPV DNA. Results: We found that 56 (36.6%) of 153 women were positive for HPV in the cervix. Logistic regression analyses showed a decrease of prevalence of HPV infection with increasing maternal age (P = .02). No HPV DNA could be detected in the amniotic fluid or cord blood, whereas eight placental specimens were positive for HPV DNA. Conclusion: The infection rate in women without clinical symptoms of HPV infection is high, but there was no HPV found in the amniotic fluid and in cord blood in women with subclinical infection in the third trimester.

Gene expression profiling of cervical tissue during physiological cervical effacement.

OBJECTIVE: The softening and dilation of cervical tissue during parturition requires a rapid reorganization of extracellular matrix and cellular interactions. The purpose of this study was to gain insight into the complex transformational changes in gene expression that lead to cervical effacement. METHODS: Cervical biopsies from effaced cervices of 10 women undergoing spontaneous vaginal delivery and from competent cervices of 10 women undergoing primary cesarean delivery were collected at 37–41 weeks of gestation and subjected to differential complementary DNA (cDNA) microarray analysis. Gene expression results were validated by real-time polymerase chain reaction (PCR). RESULTS: In a cDNA array that enables the analysis of the differential gene expression of more than 600 genes, the messenger (m)RNA expression of 40 genes increased more than 2.5-fold during cervical ripening. The majority of these genes encode cytokines, transcription factors, and cell-matrix–associated proteins. The mRNA expression of 6 genes decreased to less than 0.5-fold. The remaining 556 genes were not significantly altered. Real-time PCR analysis performed for selected, highly up-regulated genes confirmed our cDNA array findings. CONCLUSION: Complete cervical effacement is associated with a characteristic and profound alteration in the gene expression profile of cervical cells. We hypothesize that an understanding of the molecular events that accompany physiological cervical dilation is pivotal to an understanding of pathological conditions such as premature delivery and postterm pregnancy. LEVEL OF EVIDENCE: II-2

Peroxisome Proliferator-Activated REceptor γ Coactivator-1α Gene Variations Are Not Associated With Gestational Diabetes Mellitus

Objective: Epidemiologic, pathophysiologic, and genetic data suggest a close link between gestational diabetes mellitus (GDM) and type 2 diabetes. Previous studies yielded controversial results on the impact of peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1) gene variations on the development of type 2 diabetes mellitus. Therefore, we examined two common single nucleotide polymorphisms (SNP) of this gene in women with GDM. Methods: We assesed a total of 875 women by oral glucose tolerance testing (OGTT). Two hundred women of this population, 100 patients with an abnormal OGTT and 100 normal controls, were randomly slected. DNA samples isolated from the blood of the control and study groups were analyzed with respect to the SNP Gly482Ser and Thr394Thr of the PGC-1 gene using polymerase chain reaction (PCR) amplification and restriction analysis. Furthermore, a potential interaction between the Gly482Ser and the Thr394Thr variant on the risk of GDM was investigated. Results: Women with GDM were significantly older (32.2 ± 5.5 years vs 29.7 ± 6.1 years; P = .005), had higher body mass indices (BMI; 28.0 ± 7.1 kg/m2 vs 25.0 ± 5.7 kg/m2; p = .002) and displayed higher memoglobin A1c (HbA1c) values (5.6 ± 0.9 vs 4.9 vs 4.9 ± 0.5; P <.001). There was no significant difference between the allele distribution of the two polymorphisms in women with and without GDM. No significant associations between the two polymorphisms and BMI or OGTT values were observed. When the different haplotype combinations of the two loci were analyzed for the risk of GDM, on significant association could be found. Conclusion: Based on our data, the Gly-482Ser and the Thr394Thr polymorphisms of the PGC-1 gene are not associated with the development of GDM.

Transforming growth factor-beta 1 serum levels in pregnancy and pre-eclampsia

Background.u2003 Isoforms of transforming growth factor‐beta (TGF‐β1) are thought to be involved in the pathogenesis of pre‐eclampsia. Data with respect to TGF‐β1 are controversial. We examined the correlation between TGF‐β1 serum levels and the occurrence and severity of pre‐eclampsia.

Ten Polymorphisms of Estrogen-Metabolizing Genes and a Family History of Colon Cancer—An Association Study of Multiple Gene-Gene Interactions:

Estrogen replacement therapy is associated with a reduced risk of colon cancer. Therefore, we evaluated the following ten estrogen metabolism-associated single-nucleotide polymorphisms (SNPs) by sequencing-on-chip technology using solid-phase polymerase chain reaction (PCR) on oligonucleotide microarrays: catechol-O-methyltransferase (COMT) Val158Met G→A, 17-beta-hydroxysteroid dehydrogenase type 1 (HSD17) vlV A→C, cytochrome P-450 (CYP) 17 A2 allele T→C, CYP1A1 MspI RFLP f→C, CYP1A1 Ile462Val A→G, CYP19 Arg264Cys C→T, CYP19 C1558T C→T, CYP 1B1 Leu432Val, CYP1B1 Asn453Ser, and estrogen receptor (ER) alpha IVS1 - 401→C in 76 patients with a family history of colon cancer and 722 healthy controls. Using stepwise logistic regression models, we found that none of the investigated SNPs is associated with a family history of colon cancer in a univariate and multivariate logistic regression model. In addition, when all two-way interactions of the investigated SNPs were ascertained, no significant interactions between SNPs were observed. In conclusion, we found no association between the carriage of one or multiple SNPs of the estrogen metabolism and a family history of colon cancer.

β-HCG/LH Receptor (β-HCG/LH-R) Expression in Eutopic Endometrium and Endometriotic Implants: Evidence for β-HCG Sensitivity of Endometriosis

Background: Luteinizing hormone (LH) and human chorionic gonadotropin (HCG) target their receptor in gonadal and nongonadal cells to stimulate steroidogenesis and cell growth. The aim of the present study was to investigate the expression of HCG/LH-R in endometriosis to elucidate a possible impact of LH and HCG on this disease. Materials and methods: Analysis of HCG/LH-R protein expression in 23 paired samples of ectopic and eutopic tissue of cycling women with endometriosis and in endometrial samples from 22 healthy controls was conducted via immunofluorescence. HCG and HCG/LH-R gene expression in endometriotic lesions was confirmed by reverse-transcriptase polymerase chain reaction. Results: In endometriotic implants, epithelial HCG/LH-R was found in 12/23 samples. No significant differences in HCG/LH-R levels were observed when compared with glands of uterine endometrium from the same patients or healthy controls. Messenger RNA transcripts for HCG were detected in all 12 samples, whereas HCG/LH-R mRNAs were observed in 10 of the 12 endometriotic lesions investigated. Conclusions: Although HCG/LH-R was not found to be selectively upregulated in endometriosis, the mere presence of HCG/LH-R in endometriotic tissue may suggest sensitivity of endometriosis to HCG and LH that target HCG/LH-R.

An lnterleukin-6 Gene Promoter Polymorphism and Unexplained Late Intrauterine Fetal Death: A Multicenter Study

Objective: Interleukin-6 (IL-6)-mediated inflammatory processes have been proposed to be involved in the pathogenesis of pregnancy-associated complications such as late unexplained intrauterine fetal death (IUFD). Therefore we determined whether a common guanine/cytosine polymorphism at position-174 of the promoter of the IL-6 gene (1L6) known to affect in vivo protein activity can serve as candidate genefor this condition. Methods: In a multicenter case-control study, we evaluated the IL6 promoter polymorphism by pyrosequencing in 92 women with IUFD. Ninety-four healthy women with at least one uncomplicated full-term pregnancy and no history of IUFD served as the control group. Results: No significant association was found between the presence of at least one mutant allele of the IL6 promoter polymorphism (P = .2; odds ratio = 1.5 [95% confidence interval, 0.8-2.7]) and the incidence of IUFD. In women with IUFD, the presence of at least one mutant allele of the IL6 promoter polymorphism did not influence timing offetal death (33.9 [5. 1]gestational weeks vs 34. 1 [4.9]gestational weeks, P = .8) or birth weight (2055 [1119] g vs 1963 [992] g, P = .7). Conclusion: To our knowledge, we are thefirst to report on a common polymorphism of the IL6 promoter gene in women with late IUFD. The investigated IL6 promoter polymorphism can not be seen as candidate gene for IUFD in Caucasian women.

Reduction of preeclampsia in multiple pregnancies by a dedicated monitoring protocol.

Objective: To evaluate the effect of a dedicated monitoring protocol on the incidence of pregnancy-induced hypertension disorders (preeclampsia, HELLP [hemolysis, elevated liver enzymes, low platelets] syndrome, gestational hypertension) during a 5-year period in 417 women with multiple gestations. Methods: At the Department of Obstetrics and Fetomaternal Medicine, Vienna University, an outpatient care protocol for women with multiple pregnancies was established. Between March 1997 and February 2002, 379 twins and 38 triplets were followed up. Mean visits of dichorial and monochorial twins were nine and 11, respectively. Triplets were asked to at least 12 follow-up visits. Results: Any kind of pregnancy-associated hypertensive disorders necessitating cesarean delivery developed in eight women (seven with twins, one with triplets; 1.76%). Three cases of preeclampsia, three cases of HELLP syndrome, and two cases of pregnancy-induced hypertension were diagnosed. Perinatal outcome of all 17 newborns was excellent without any mortality and only minor morbidity. All mothers left the hospital in a fully recovered condition. Conclusion: Because of our results we hypothesize that our monitoring protocol with frequent visits, continuous personal obstetric care, and timed delivery is effective in reduction of pregnancy-induced hypertension in multiple pregnancies.