Interstitial collagen gives fetal membranes tensile strength, and membrane rupture has been attributed to collagen degradation. A polymorphism at −1607 in the matrix metalloproteinase-1 (MMP-1) ...promoter (an insertion of a guanine (G)) creates a core Ets binding site and increases promoter activity. We investigated whether this polymorphism is functionally significant for MMP-1 expression in amnion cells and whether it is associated with preterm premature rupture of the membranes (PPROM). The 2G promoter had >2-fold greater activity than the 1G allele in amnion mesenchymal cells and WISH amnion cells. Phorbol 12-myristate 13-acetate (PMA) increased mesenchymal cell nuclear protein binding with greater affinity to the 2G allele. Induction of MMP-1 mRNA by PMA was significantly greater in cells with a 1G/2G or 2G/2G genotype compared with cells homozygous for the 1G allele. When treated with PMA, the 1G/2G and 2G/2G cells produced greater amounts of MMP-1 protein than 1G/1G cells. A significant association was found between fetal carriage of a 2G allele and PPROM. We conclude that the 2G allele has stronger promoter activity in amnion cells, that it confers increased responsiveness of amnion cells to stimuli that induce MMP-1, and that this polymorphism contributes to the risk of PPROM.
Two cohorts of women with polycystic ovary syndrome (PCOS), comprising 400 probands and affected sisters in 365 families and a case-control group including 395 women with PCOS and 171 healthy women ...with regular menstrual cycles, were studied to determine whether single-nucleotide polymorphisms (SNPs) identified as susceptibility loci in genomewide association studies of type 2 diabetes are also associated with PCOS. None of the 18 allelic variants in 10 genes previously shown to be associated with type 2 diabetes were found to be associated with PCOS, but some were associated with indices of beta cell function.
Since mediators of inflammation are associated with insulin resistance, and the risk of developing diabetes mellitus and gestational diabetes, we hypothesized that genetic variation in members of the ...inflammatory gene pathway impact glucose levels and related phenotypes in pregnancy. We evaluated this hypothesis by testing for association between genetic variants in 31 inflammatory pathway genes in the Hyperglycemia and Adverse Pregnancy Outcome (HAPO) cohort, a large multiethnic multicenter study designed to address the impact of glycemia less than overt diabetes on pregnancy outcome.
Fasting, 1-hour, and 2-hour glucose, fasting and 1-hour C-peptide, and HbA1c levels were measured in blood samples obtained from HAPO participants during an oral glucose tolerance test at 24-32 weeks gestation. We tested for association between 458 SNPs mapping to 31 genes in the inflammatory pathway and metabolic phenotypes in 3836 European ancestry and 1713 Thai pregnant women. The strongest evidence for association was observed with TNF alpha and HbA1c (rs1052248; 0.04% increase per allele C; p-value = 4.4×10(-5)), RETN and fasting plasma glucose (rs1423096; 0.7 mg/dl decrease per allele A; p-value = 1.1×10(-4)), IL8 and 1 hr plasma glucose (rs2886920; 2.6 mg/dl decrease per allele T; p-value = 1.3×10(-4)), ADIPOR2 and fasting C-peptide (rs2041139; 0.55 ug/L decrease per allele A; p-value = 1.4×10(-4)), LEPR and 1-hour C-peptide (rs1171278; 0.62 ug/L decrease per allele T; p-value = 2.4×10(-4)), and IL6 and 1-hour plasma glucose (rs6954897; -2.29 mg/dl decrease per allele G, p-value = 4.3×10(-4)).
Based on the genes surveyed in this study the inflammatory pathway is unlikely to have a strong impact on maternal metabolic phenotypes in pregnancy although variation in individual members of the pathway (e.g. RETN, IL8, ADIPOR2, LEPR, IL6, and TNF alpha,) may contribute to metabolic phenotypes in pregnant women.
Nature Communications 6: Article number: 7502 (2015); Published: 18 August 2015; Updated: 12 February 2016 In the original version of this Article the genetic locus 8p23.1 was incorrectly referred to ...as 8p32.1 throughout. For example, in the Abstract, the sentence beginning ‘Three loci reach genome-wide significance in the case-control meta-analysis…' originally read ‘Three loci reach genome-wide significance in the case-control meta-analysis, two novel loci mapping to chr 8p32.
The androgen receptor (AR) is important in reproductive organ development, as well as tissue homeostasis of the pancreas, liver, and skeletal muscle in adulthood. The trinucleotide (CAG)n repeat ...polymorphism in exon 1 of the AR gene is thought to regulate AR activity, with longer alleles conferring reduced receptor activity. Therefore, the evaluation of the allelic distribution of the AR (CAG)n repeat in various ethnic groups is crucial in understanding the interindividual variability in AR activity. We evaluated ethnic variation of this AR polymorphism by genotyping individuals from the multiethnic Hyperglycemia and Adverse Pregnancy Outcome study cohort. We genotyped 4421 Caucasian mothers and 3365 offspring of European ancestry; 1494 Thai mothers and 1742 offspring; 1119 Afro‐Caribbean mothers and 1142 offspring; and 780 Hispanic mothers and 770 offspring of Mexican ancestry from Bellflower, California. The distributions of (CAG)n alleles among all 4 ethnic groups are significantly different (P < .0001). Pairwise tests confirmed significant differences between each pair of ethnicities tested (P < 10−28). The relative AR (CAG)n repeat length in the different groups was as follows: Afro‐Caribbean (shortest repeat lengths and greatest predicted AR activity) < Caucasian < Hispanic < Thai (longest repeat length and lowest predicted AR activity). Significant interethnic differences in the allele frequencies of the AR exon 1 (CAG)n polymorphism exist. Our results suggest that there may be potential ethnic differences in androgenic pathway activity and androgen sensitivity.
Abstract
Context
Polycystic ovary syndrome (PCOS) is among the most common endocrine disorders of premenopausal women, affecting 5% to15% of this population depending on the diagnostic criteria ...applied. It is characterized by hyperandrogenism, ovulatory dysfunction, and polycystic ovarian morphology. PCOS is highly heritable, but only a small proportion of this heritability can be accounted for by the common genetic susceptibility variants identified to date.
Objective
The objective of this study was to test whether rare genetic variants contribute to PCOS pathogenesis.
Design, Patients, and Methods
We performed whole-genome sequencing on DNA from 261 individuals from 62 families with one or more daughters with PCOS. We tested for associations of rare variants with PCOS and its concomitant hormonal traits using a quantitative trait meta-analysis.
Results
We found rare variants in DENND1A (P = 5.31 × 10−5, adjusted P = 0.039) that were significantly associated with reproductive and metabolic traits in PCOS families.
Conclusions
Common variants in DENND1A have previously been associated with PCOS diagnosis in genome-wide association studies. Subsequent studies indicated that DENND1A is an important regulator of human ovarian androgen biosynthesis. Our findings provide additional evidence that DENND1A plays a central role in PCOS and suggest that rare noncoding variants contribute to disease pathogenesis.
Whole-genome sequencing analysis finds rare variants in associated with altered hormone levels in families affected by PCOS.
Abstract
Context
Polycystic ovary syndrome (PCOS) is a highly heritable, common endocrine disorder characterized by hyperandrogenism, irregular menses, and polycystic ovaries. PCOS is often ...accompanied by elevated levels of anti-Müllerian hormone (AMH). AMH inhibits follicle maturation. AMH also inhibits steroidogenesis through transcriptional repression of CYP17A1. We recently identified 16 rare PCOS-specific pathogenic variants in AMH.
Objective
To test whether additional members of the AMH signaling pathway also contribute to the etiology of PCOS.
Participants/Design
Targeted resequencing of coding and regulatory regions of AMH and its specific type 2 receptor, AMHR2, was performed on 608 women affected with PCOS and 142 reproductively normal control women. Prediction tools of deleteriousness and in silico evidence of epigenetic modification were used to prioritize variants for functional evaluation. Dual-luciferase reporter assays and splicing assays were used to measure the impact of genetic variants on function.
Results
We identified 20 additional variants in/near AMH and AMHR2 with significantly reduced signaling activity in in vitro assays. Collectively, from our previous study and as reported herein, we have identified a total of 37 variants with impaired activity in/near AMH and AMHR2 in 41 women affected with PCOS, or 6.7% of our PCOS cohort. Furthermore, no functional variants were observed in the 142 phenotyped controls. The functional variants were significantly associated with PCOS in our cohort of 608 women with PCOS and 142 controls (P = 2.3 × 10−5) and very strongly associated with PCOS relative to a larger non-Finnish European (gnomAD) population-based control cohort (P < 1 × 10−9).
Conclusion
The AMH signaling cascade plays an important role in PCOS etiology.
Functional characterization of genetic variation in AMH and its cognate receptor, AMHR2, in women with PCOS provides strong evidence for the role of impaired AMH signaling in the pathology of PCOS.
Abstract
Context
Polycystic ovary syndrome (PCOS) is a prevalent disorder in reproductive aged women associated with a number of endocrine and metabolic complications, including increased risk of ...endometrial cancer.
Objective
To study the effect of the characteristic increased androgen levels in PCOS on the endometrium, a novel scaffold-free multicellular endometrial organoid was established.
Design
Human endometrial organoids were constructed using primary endometrial epithelial and stromal cells from endometrial tissues. Organoids were treated for 14 days with physiologic levels of estradiol and testosterone to mimic a normal follicular phase or PCOS hormone profiles. Organoids were harvested for immunostaining and ribonucleic acid sequencing.
Setting
Academic institution.
Patients
Endometrial tissues from 10 premenopausal women undergoing hysterectomy for benign pathologies were obtained following written consent.
Main Outcome Measures
Organoid architecture, cell specific markers, functional markers, proliferation, and gene expression were measured.
Results
A method to generate scaffold-free endometrial organoids containing epithelial and stromal cells was established. These organoids exhibited distinct organization with epithelial cells lining the outer surface and stromal cells in the center of the organoids. Epithelial cells were polarized, organoids expressed cell type specific and functional markers, as well as androgen, estrogen, and progesterone receptors. Treatment with PCOS hormones increased cell proliferation and dysregulated genes in endometrial organoids.
Conclusions
A new multicellular, scaffold-free endometrial organoid system was established that resembled physiology of the native endometrium. Excess androgens in PCOS promoted cell proliferation in endometrial organoids, revealing new mechanisms of PCOS-associated with risk of endometrial neoplasia.
Variation in Resistin Gene Promoter Not Associated With Polycystic Ovary Syndrome
Margrit Urbanek 1 ,
Yangzhu Du 1 ,
Kaisa Silander 2 ,
Francis S. Collins 2 ,
Claire M. Steppan 3 ,
Jerome F. Strauss ...III 4 ,
Andrea Dunaif 5 ,
Richard S. Spielman 1 and
Richard S. Legro 6
1 Department of Genetics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania
2 Genome Technology Branch, National Human Genome Research Institute, Bethesda, Maryland
3 Department of Medicine, Division of Endocrinology, Diabetes, and Metabolism, University of Pennsylvania School of Medicine,
Philadelphia, Pennsylvania
4 Center for Research on Reproduction and Women’s Health and Department of Obstetrics and Gynecology, University of Pennsylvania
School of Medicine, Philadelphia, Pennsylvania
5 Division of Endocrinology, Metabolism and Molecular Medicine, Northwestern University Medical School, Chicago, Illinois
6 Department of Obstetrics and Gynecology, College of Medicine, Pennsylvania State University, Hershey, Pennsylvania
Abstract
Polycystic ovary syndrome (PCOS) is a leading cause of anovulatory infertility and affects ∼4–7% of reproductive age women
in the U.S. It is characterized by hyperandrogenemia and chronic anovulation and is associated with insulin resistance, obesity,
and increased risk for type 2 diabetes. In a screen of candidate genes, a region on chromosome 19p13.3 was identified that
shows significant evidence for both linkage and association with PCOS. A promising candidate gene for PCOS, resistin, maps
to exactly this region. Resistin is a protein hormone thought to modulate glucose tolerance and insulin action. We tested
for association between a single nucleotide polymorphism in the promoter region of the resistin gene and three phenotypes:
PCOS, obesity, and insulin resistance. We did not find evidence for association with any of the phenotypes. It is therefore
unlikely that variation in the resistin gene accounts for the strong association that we observe between chromosome 19p13.3
and PCOS. Instead, this association is most likely due to a gene or genetic element in this region that has not been identified.
Footnotes
Address correspondence and reprint requests to Richard S. Spielman, Department of Genetics, Clinical Research Building, University
of Pennsylvania School of Medicine, 415 Curie Blvd., Philadelphia, PA 19104-6145. E-mail: spielman{at}pobox.upenn.edu .
Received for publication 7 June 2002 and accepted in revised form 14 October 2002.
M.U. is currently located at the Department of Medicine, Division of Endocrinology, Metabolism, and Molecular Medicine, Northwestern
University Medical School, Chicago, Illinois.
HA, hyperandrogenemia; PCOS, polycystic ovary syndrome; PPARγ, peroxisome proliferator–activated receptor-γ; SNP, single nucleotide
polymorphism; TDT, transmission/disequilibrium test.
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