Dr. Aghajanova received her medical degree from Yerevan State Medical University in Armenia, followed by residency in obstetrics and gynecology, then completed PhD in Human Implantation at Karolinska Institute, Stockholm, Sweden, followed with embryology training at Karolinska Institute, with an Internship in Austria.
Subsequently, Dr. Aghajanova completed residency in obstetrics and gynecology at Baylor College of Medicine, Houston, Texas and at UC San Francisco with subsequent Reproductive Endocrinology and Infertility fellowship training at UC San Francisco. She is a respected researcher in the field of endometrial receptivity, implantation and endometriosis.
Dr. Aghajanova speaks Russian and Armenian and is very well published with over 50 peer-reviewed publications as well as numerous other oral and poster presentations and is a professional peer-reviewer for over 12 journals.
Dr.Aghajanova enjoys spending time with her husband and children, and traveling.
- Obstetrics and Gynecology
- Reproductive Endocrinology & Infertility
- Endometrial aspects of fertility
Clinical Assistant Professor, Obstetrics & Gynecology - Reproductive Endocrinology & Infertility
Residency: Baylor College of Medicine Obstetrics and Gynecology Residency (2012) TX
Fellowship: UCSF Reproductive Endocrinology and Infertility Fellowship (2017) CA
Residency: UCSF Obstetrics and Gynecology Residency (2014) CA
Board Certification: Obstetrics and Gynecology, American Board of Obstetrics and Gynecology (2017)
PhD, Karolinska Institue, Sweden (2006)
Residency, Yerevan State Medical University, Armenia, Obstetrics & Gynecology (1999)
MD, Yerevan State Medical University, Armenia, Medical Doctor (1996)
Platelet Rich Plasma for Patients With Recurrent Implantation Failure
Patients with recurrent implantation failure are among the most difficult patients to treat, with no proven standard treatment. Platelet rich plasma stimulates cellular processes involved in endometrial regeneration, and in a small case series has shown efficacy for this patient population. We hope to conduct a randomized controlled pilot study to determine whether PRP is indeed an effective treatment for recurrent implantation failure.
- Should we consider alternative therapies to operative hysteroscopy for the treatment of Asherman syndrome? Fertility and sterility 2020
- Correction to: How good are we at modeling implantation? Journal of assisted reproduction and genetics 2020
Cross-disorder analysis of endometriosis and its comorbid diseases reveals shared genes and molecular pathways and proposes putative biomarkers of endometriosis.
Reproductive biomedicine online
RESEARCH QUESTION: Women with endometriosis are considered to be at higher risk of several chronic diseases, such as autoimmune disorders, gynaecological cancers, asthma/atopic diseases and cardiovascular and inflammatory bowel diseases. Could the study of endometriosis-associated comorbidities help to identify potential biomarkers and target pathways of endometriosis?DESIGN: A systematic review was performed to identify all possible endometriosis-associated comorbid conditions. Next, this list of disorders was coded into MeSH terms, and the gene expression profiles were downloaded from the Phenopedia database and subsequently analysed following a systems biology approach.RESULTS: The results identified a group of 127 candidate genes that were recurrently expressed in endometriosis and its closest comorbidities and that were defined as 'endometriosis sibling disorders' (ESD). The enrichment analysis showed that these candidate genes are principally involved in immune and drug responses, hormone metabolism and cell proliferation, which are well-known hallmarks of endometriosis. The expression of ESD genes was then validated on independent sample cohorts (n = 207 samples), in which the involvement of 16 genes (AGTR1, BDNF, C3, CCL2, CD40, CYP17A1, ESR1, IGF1, IGF2, IL10, MMP1, MMP7, MMP9, PGR, SERPINE1 and TIMP2) in endometriosis was confirmed. Several of these genes harbour polymorphisms that associate to either endometriosis or its comorbid conditions.CONCLUSIONS: The study results highlight the molecular processes underlying the aetiopathogenesis of endometriosis and its comorbid conditions, and identify putative endometriosis biomarkers.
View details for DOI 10.1016/j.rbmo.2019.11.003
View details for PubMedID 31926826
Levonorgestrel IUD: is there a long-lasting effect on return to fertility?
Journal of assisted reproduction and genetics
Intrauterine devices (IUDs) are effective and safe long-acting reversible contraceptive methods for preventing unplanned pregnancies. While extensive studies were conducted to evaluate return to fertility after removal of IUDs, majority of them were focused on multiparous women using copper IUDs. Current trends indicate increased use of levonorgestrel (LNG) IUDs in nulliparous women for very long periods of time, with both nulliparity and long duration of LNG-IUD use being potentially associated with trends towards longer time to conception post removal. Understanding the effects that LNG-IUDs may have on endometrial morphology and gene expression has important implications to further understanding their mechanism of action. Studies examining endometrial gene expression show persistent changes in receptivity markers up to 1 year after removal of an inert IUD, and no similar studies have been performed after removal of LNG-IUDs. Given the current gap in the literature and trends in LNG-IUD use in nulliparous young women, studies are needed that specifically look at the interaction of nulliparity, long-term use of LNG-IUD, and return to normal fertility. Herein, we review the available literature on the mechanism of action of IUDs with a specific focus on the effect on endometrial gene expression profile changes associated with IUDs.
View details for DOI 10.1007/s10815-019-01624-5
View details for PubMedID 31709489
- Growth Hormone and Endometrial Receptivity FRONTIERS IN ENDOCRINOLOGY 2019; 10
- PREGNANCY SUCCESS RATES FOR LESBIAN COUPLES UNDERGOING INTRAUTERINE INSEMINATION. ELSEVIER SCIENCE INC. 2019: E61
- NATIONWIDE SURVEY OF ACCESS TO CARE INITIATIVES IN REI PRACTICES ASSOCIATED WITH OB/GYN RESIDENCY PROGRAMS. ELSEVIER SCIENCE INC. 2019: E111
- CLINICAL EXPOSURE IN OB/GYN RESIDENT TRAINING PROGRAMS IN THE UNITED STATES TO INFERTILITY CARE FOR LOW RESOURCE AND UNDERSERVED COMMUNITIES. ELSEVIER SCIENCE INC. 2019: E405–E406
- INFERTILITY, FERTILITY PRESERVATION, AND ACCESS TO CARE DURING TRAINING: A NATION-WIDE MULTI-SPECIALTY SURVEY OF UNITED STATES RESIDENTS AND FELLOWS. ELSEVIER SCIENCE INC. 2019: E407
- EFFECT OF ESTRACE ON PREGNANCY RATES FOR WOMEN WITH THIN ENDOMETRIAL LINING UNDERGOING INTRAUTERINE INSEMINATION. ELSEVIER SCIENCE INC. 2019: E335
- Platelet-rich plasma in the management of Asherman syndrome: case report. Journal of assisted reproduction and genetics 2018
In vitro evidence that platelet-rich plasma stimulates cellular processes involved in endometrial regeneration.
Journal of assisted reproduction and genetics
The study aims to test the hypothesis that platelet-rich plasma (PRP) stimulates cellular processes involved in endometrial regeneration relevant to clinical management of poor endometrial growth or intrauterine scarring.Human endometrial stromal fibroblasts (eSF), endometrial mesenchymal stem cells (eMSC), bone marrow-derived mesenchymal stem cells (BM-MSC), and Ishikawa endometrial adenocarcinoma cells (IC) were cultured with/without 5% activated (a) PRP, non-activated (na) PRP, aPPP (platelet-poor-plasma), and naPPP. Treatment effects were evaluated with cell proliferation (WST-1), wound healing, and chemotaxis Transwell migration assays. Mesenchymal-to-epithelial transition (MET) was evaluated by cytokeratin and vimentin expression. Differential gene expression of various markers was analyzed by multiplex Q-PCR.Activated PRP enhanced migration of all cell types, compared to naPRP, aPPP, naPPP, and vehicle controls, in a time-dependent manner (p < 0.05). The WST-1 assay showed increased stromal and mesenchymal cell proliferation by aPRP vs. naPRP, aPPP, and naPPP (p < 0.05), while IC proliferation was enhanced by aPRP and aPPP (p < 0.05). There was no evidence of MET. Expressions of MMP1, MMP3, MMP7, and MMP26 were increased by aPRP (p < 0.05) in eMSC and eSF. Transcripts for inflammation markers/chemokines were upregulated by aPRP vs. aPPP (p < 0.05) in eMSC and eSF. No difference in estrogen or progesterone receptor mRNAs was observed.This is the first study evaluating the effect of PRP on different human endometrial cells involved in tissue regeneration. These data provide an initial ex vivo proof of principle for autologous PRP to promote endometrial regeneration in clinical situations with compromised endometrial growth and scarring.
View details for DOI 10.1007/s10815-018-1130-8
View details for PubMedID 29404863
Obstetrics and Gynecology Residency and Fertility Needs.
2017; 24 (3): 428-434
Infertility is a common reproductive disease, with a prevalence of 9% to 18% of the general population. To date, no studies have attempted to examine the prevalence and experience of infertility among resident physicians in the United States. In female obstetrics and gynecology (Ob/Gyn) residents of age where infertility becomes more prevalent, ability to seek fertility may be influenced by rigorous professional demands and low remuneration. We seek to understand the prevalence of infertility, as well as experience and utilization of infertility services among Ob/Gyn residents. Cross-sectional descriptive survey was distributed among US Accreditation Council for Graduate Medical Education-accredited Ob/Gyn programs. Demographics, intentions to conceive during residency, fertility problems, fertility treatment, affordability of care, and perceptions of support were surveyed. A total of 241 responses were received in an equal distribution between junior (n = 120) and senior (n = 121) residents. The majority of respondents were female (91%), 25 to 35 years old (94%), and married (54%). Eighty-five percent (195 of 230) did not actively pursue fertility during residency. Twenty-nine percent (68 of 235) considered fertility preservation, but only 2% sought consultation. Twenty-nine percent of those interested in fertility (22 of 75) experienced infertility of some degree. Sixty-three percent felt low or no support from the program. Thirty-five percent reported stigma associated with their infertility. In conclusion, infertility is a prevalent reproductive health impairment among Ob/Gyn residents. The majority of residents defer childbearing during residency despite advancing reproductive age. A majority felt little or no support from training programs in addressing their fertility care. Further studies are indicated to understand the barriers and impact among resident trainees.
View details for DOI 10.1177/1933719116657193
View details for PubMedID 27368879
Effects of noncavity-distorting fibroids on endometrial gene expression and function.
Biology of reproduction
2017; 97 (4): 564–76
Uterine fibroids are a common finding in infertility patients. Impaired implantation and decidualization have been proposed to contribute to compromised fertility. Data are limited on the endometrial transcriptome from subjects with uterine fibroids, as well as endometrial receptivity and decidualization potential of endometrial stromal fibroblasts (eSF) from women with fibroids. Our objective was to investigate the endometrial transcriptome of women with noncavity-distorting intramural fibroids and compare them to control subjects with no uterine pathology throughout the menstrual cycle. We also evaluated endometrial receptivity gene expression and basic endometrial functions such as decidualization, proliferation, and apoptosis in women with fibroid uterus. Results showed that large numbers of transcripts were significantly dysregulated throughout the menstrual cycle in fibroid subjects compared to controls. However, there were essentially no differences in the expression of receptivity markers at the tissue level, as well as decidualization markers in tissue and eSF in subjects with fibroids compared to controls. However, eSF from women with a fibroid uterus exhibited decreased proliferation potential and increased apoptosis upon decidualization. These data indicate preserved implantation and decidualization potential despite observed gene expression changes in endometrium from women with noncavity-distorting fibroids compared to controls. How this phenomenon and altered proliferation/apoptosis may contribute to impairment of endometrial function in subfertile patients warrants further investigation.
View details for DOI 10.1093/biolre/iox107
View details for PubMedID 29025102
Meta-signature of human endometrial receptivity: a meta-analysis and validation study of transcriptomic biomarkers.
2017; 7 (1): 10077
Previous transcriptome studies of the human endometrium have revealed hundreds of simultaneously up- and down-regulated genes that are involved in endometrial receptivity. However, the overlap between the studies is relatively small, and we are still searching for potential diagnostic biomarkers. Here we perform a meta-analysis of endometrial-receptivity associated genes on 164 endometrial samples (76 from 'pre-receptive' and 88 from mid-secretory, 'receptive' phase endometria) using a robust rank aggregation (RRA) method, followed by enrichment analysis, and regulatory microRNA prediction. We identify a meta-signature of endometrial receptivity involving 57 mRNA genes as putative receptivity markers, where 39 of these we confirm experimentally using RNA-sequencing method in two separate datasets. The meta-signature genes highlight the importance of immune responses, the complement cascade pathway and the involvement of exosomes in mid-secretory endometrial functions. Bioinformatic prediction identifies 348 microRNAs that could regulate 30 endometrial-receptivity associated genes, and we confirm experimentally the decreased expression of 19 microRNAs with 11 corresponding up-regulated meta-signature genes in our validation experiments. The 57 identified meta-signature genes and involved pathways, together with their regulatory microRNAs could serve as promising and sought-after biomarkers of endometrial receptivity, fertility and infertility.
View details for DOI 10.1038/s41598-017-10098-3
View details for PubMedID 28855728
View details for PubMedCentralID PMC5577343
Demographic analysis of a low resource, socioculturally diverse urban community presenting for infertility care in a United States public hospital.
Contraception and reproductive medicine
2017; 2: 17
Infertility is a prevalent disease of reproductive health that exerts an impact on an estimated 80 million people worldwide. For many, involuntary childlessness becomes a central and preoccupying issue in their lives, the impact of which is exacerbated by lack of access to basic care and treatment. These effects maybe further magnified among immigrant communities, a growing but highly marginalized population that has been shown in other areas of reproductive health to experience worse health outcomes and delays in access to care. To date, few studies have examined the unique medical and sociocultural considerations of infertility among immigrant populations in the United States.Our study is a cross-sectional analysis of women presenting for infertility evaluation at a county hospital serving a low resource, socioculturally diverse largely immigrant communities in comparison to infertile women from a largely affluent population presenting to a high resource, comprehensive fertility center. We employed surveys to evaluate demographics and socioeconomic parameters as well as abstracted data from medical records to obtain infertility diagnoses. Multivariate regression analysis was applied to examine impact of sociocultural factors as predictors of duration of untreated infertility disease burden experienced by patients.Eighty-seven women were included in our analysis. In the county hospital/low resource clinic (LR), the mean age was 32.9 ± 4.9 vs 36.4 ± 6.3 years in the fee-for-service/high resource clinic (HR). The mean reported duration of infertility in LR and HR patients was 3.4 ± 3.0 vs 2.3 ± 1.5 years. 70% of LR patients were monolingual non-English speakers vs 5.4% of HR patients. 59% of LR patients reported an annual household income of less than $25,000 and 70% did not have a college degree. 81.1% of HR patients reported an income of higher than $100,000, and 81.1% had completed college or graduate school. The most common infertility diagnosis in the LR was anovulation (38%) and tubal factor (28%) compared to diminished ovarian reserve (37.8%) and male factor (51.4%) in the HR. After controlling for age at the initiation of pregnancy attempt, lower education level, lower income, and immigrant status were significantly correlated with a longer duration of infertility.Women presenting for infertility care to a low resource county medical center represent immigrant communities and are generally of younger age, but with a longer duration of infertility. This study identifies lower educational level, income, and immigrant status as barriers in access to care.
View details for DOI 10.1186/s40834-017-0044-7
View details for PubMedID 29201422
View details for PubMedCentralID PMC5683225
Global Transcriptome Abnormalities of the Eutopic Endometrium From Women With Adenomyosis
2016; 23 (10): 1289-1303
Adenomyosis is a clinical disorder defined by the presence of endometrial glands and stroma within the myometrium, the pathogenesis of which is poorly understood. We postulate that dysregulation of genes and pathways in eutopic endometrium may predispose to ectopic implantation. No study, to our knowledge, has examined the global transcriptome of isolated eutopic endometrium from women with clinically significant adenomyosis.Laboratory-based study with full institutional review board approval and consents.Endometrial sampling was performed on hysterectomy specimens (proliferative phase) from symptomatic women with pathologically confirmed diffuse adenomyosis (n = 3). Controls (n = 5) were normo-ovulatory patients without adenomyosis. All patients were free from leiomyoma, endometriosis, and hormonal exposures. Isolated purified total RNA was subjected to microarray analysis using the Gene 1.0 ST Affymetrix platform. Data were analyzed with GeneSpring and Ingenuity Pathway analysis. Validation of several genes was undertaken by quantitative real-time reverse transcriptase polymerase chain reaction.Comparison of transcriptomes of proliferative endometrium from women with and without adenomyosis revealed 140 upregulated and 884 downregulated genes in samples from women with adenomyosis compared to controls. Highly differentially expressed genes include those involved in regulation of apoptosis, steroid hormone responsiveness, and proteins involved in extracellular matrix remodeling as well as microRNAs of unknown significance. Affected canonical pathways included eukaryotic initiation factor 2 signaling, oxidative phosphorylation, mitochondrial dysfunction, estrogen receptor signaling, and mammalian target of rapamycin signaling.The eutopic endometrium in patients with adenomyosis has fundamental abnormalities that may predispose to invasion and survival beyond the myometrial interface.
View details for DOI 10.1177/1933719116650758
View details for Web of Science ID 000383388400003
View details for PubMedID 27233751
Stanniocalcin-1 expression in normal human endometrium and dysregulation in endometriosis
FERTILITY AND STERILITY
2016; 106 (3): 681-?
To determine expression of stanniocalcin-1 (STC1) in human endometrium with and without endometriosis and its regulation by steroid hormones.Laboratory study.University.Nineteen women with endometriosis and 33 control women.Endometrial biopsy and fluid sampling.Analysis of early secretory (ESE) and midsecretory (MSE) endometrial secretomes from fertile women with the use of nano-liquid chromatography-dual mass spectrometry; real-time quantitative polymerase chain reaction, and immunohistochemistry for STC1 and its receptor calcium-sensing receptor (CASR) mRNA and proteins in endometrium with and without endometriosis; evaluation of STC1 and CASR mRNA expression in endometrial stromal fibroblasts (eSF) from women with and without endometriosis decidualized with the use of E2P or 8-bromo-cyclic adenosine monophosphate (cAMP).STC1 protein was strongly up-regulated in MSE versus ESE in endometrial fluid of fertile women. STC1 mRNA significantly increased in MSE from women with, but not from those without, endometriosis, compared with proliferative endometrium or ESE, with no significant difference throughout the menstrual cycle between groups. STC1 mRNA in eSF from control women increased >230-fold on decidualization with the use of cAMP versus 45-fold from women with endometriosis, which was not seen on decidualization with E2/P. CASR mRNA did not exhibit significant differences in any condition and was not expressed in isolated eSF. STC1 protein immunoexpression in eSF was significantly lower in women with endometriosis compared with control women.STC1 protein is significantly up-regulated in MSE endometrial fluid and is dysregulated in eutopic endometrial tissue from women with endometriosis. It is likely regulated by cAMP and may be involved in the pathogenesis of decidualization defects.
View details for DOI 10.1016/j.fertnstert.2016.05.023
View details for Web of Science ID 000386569800034
View details for PubMedID 27322879
View details for PubMedCentralID PMC5010972
- Birth of a healthy child after pre-implantation genetic screening of embryos from sperm of a man with nonmosaic Down syndrome JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 2016; 33 (5): 675-675
Impact of pituitary suppression on antral follicle count and oocyte recovery after ovarian stimulation
FERTILITY AND STERILITY
2016; 105 (3): 690-696
To investigate the potential influence of short-term pituitary suppression on antral follicle count (AFC) and correlate the AFC with the number of oocytes retrieved after ovarian stimulation.Retrospective study.University fertility center.A total of 1,479 infertile patients.Patients had baseline AFC, when they were not on any medications known to cause pituitary suppression, and follow-up AFC (suppressed AFC) while on E2, GnRH agonist (GnRH-a), oral contraceptive (OC) pills, or OC pills/GnRH-a in preparation for ovarian stimulation, performed within 6 months of initial baseline AFC.The AFC at baseline, AFC during pituitary suppression, and the number of oocytes retrieved.Although there was an average unadjusted decline of 0.4, 0.9, 2.2, and 3.0 in AFC while patients were on E2, GnRH-a, OC pills, and OC pills/GnRH-a, respectively, this decline was driven by age, baseline AFC, and the hormones used. Although baseline and suppressed AFC were found to be good predictors of the number of oocytes retrieved after ovarian stimulation, statistically, suppressed AFC was found to be a marginally better predictor.Short-term pituitary suppression has a negative impact on AFC. This decline in AFC may influence the number of oocytes retrieved, suggesting the suppressive impact of exogenous hormones on the biological capacity of the ovary during stimulation.
View details for DOI 10.1016/j.fertnstert.2015.11.033
View details for Web of Science ID 000373406300023
View details for PubMedID 26696299
Prepubertal Gynecomastia Due to Indirect Exposure to Nonformulary Bioidentical Hormonal Replacement Therapy A Case Report
JOURNAL OF REPRODUCTIVE MEDICINE
2016; 61 (1-2): 73-77
Gynecomastia is a disorder of the endocrine system characterized by an abnormal presence of a palpable unilateral or bilateral enlargement and proliferation of glandular ductal benign breast tissue in male individuals. This case discusses the medical implications of an unregulated, indirect exposure to nonformulary, bioidentical hormone replacement therapy in male children.An 8-year-old boy presented with prepubertal gynecomastia secondary to estrogen exposure from maternal use of bioidentical hormonal replacement therapy (the Wiley protocol). We review the literature on prepubertal gynecomastia secondary to exogenous estrogen exposure, evaluation, clinical surveillance of the pubertal development, and relevant short- and long-term implications.Indirect exposure to nonformulary hormonal replacement in our case report was an etiologic factor in the development of prepubertal gynecomastia. This novel estrogen exposure source has important implications in the differential diagnosis of prepubertal gynecomastia and potential adverse effects secondary to precocious hormonal exposure.
View details for Web of Science ID 000370116500014
View details for PubMedID 26995893
- What do we know about endometrial receptivity in women with endometriosis? A molecular perspective REPRODUCTIVE BIOMEDICINE ONLINE 2015; 31 (5): 581-583
Birth of a healthy child after preimplantation genetic screening of embryos from sperm of a man with non-mosaic Down syndrome
JOURNAL OF ASSISTED REPRODUCTION AND GENETICS
2015; 32 (9): 1409-1413
The purpose of this study is to present a case of healthy infant born after intracytoplasmic sperm injection-in vitro fertilization (ICSI-IVF) with preimplantation genetic screening (PGS) using sperm from a man with non-mosaic trisomy 21 and a literature review.A 26-year-old euploid female and 29-year-old male with non-mosaic trisomy 21 and male factor undergoing ICSI-IVF treatment for primary infertility with embryo biopsy for PGS with comprehensive chromosomal screening (CCS) presented to the Infertility Clinic at Highland Hospital, the Alameda County Medical Center, California, with 6-year history of primary infertility. The outcome measure is a live birth of a healthy child and ploidy status of biopsied blastocysts.Egg retrieval yielded 33 oocytes, 29 of which underwent ICSI with ejaculated sperm. Twenty-eight 2PN zygotes were cultured, and 13 blastocysts underwent trophectoderm biopsy and vitrification 5 or 6 days after retrieval. CCS analysis revealed that 12 out of 13 (92 %) of blastocysts were euploid and one was a complex abnormal mosaic. Transfer of two grade I hatching blastocysts resulted in a singleton pregnancy with normal prenatal genetic screening and delivery of a healthy male infant at 41 weeks via primary cesarean section for non-reassuring fetal status.This is the first report of a live birth of a healthy child after ICSI-IVF with PGS using ejaculated sperm from a man with non-mosaic trisomy 21 and male factor infertility.
View details for DOI 10.1007/s10815-015-0525-z
View details for Web of Science ID 000362519600016
View details for PubMedID 26139158
View details for PubMedCentralID PMC4595396
No evidence for mutations in NLRP7, NLRP2 or KHDC3L in women with unexplained recurrent pregnancy loss or infertility
2015; 30 (1): 232-238
Are mutations in NLRP2/7 (NACHT, LRR and PYD domains-containing protein 2/7) or KHDC3L (KH Domain Containing 3 Like) associated with recurrent pregnancy loss (RPL) or infertility?We found no evidence for mutations in NLRP2/7 or KHDC3L in unexplained RPL or infertility.Mutations in NLRP7 and KHDC3L are known to cause biparental hydatidiform moles (BiHMs), a rare form of pregnancy loss. NLRP2, while not associated with the BiHM pathology, is known to cause recurrent Beckwith Weidemann Syndrome (BWS).Ninety-four patients with well characterized, unexplained infertility were recruited over a 9-year period from three IVF clinics in Sweden. Blood samples from 24 patients with 3 or more consecutive miscarriages of unknown etiology were provided by the Recurrent Miscarriage Clinic at St Mary's Hospital, London, UK.Patients were recruited into both cohorts following extensive clinical studies. Genomic DNA was isolated from peripheral blood and subject to Sanger sequencing of NLRP2, NLRP7 and KHDC3L. Sequence electropherograms were analyzed by Sequencher v5.0 software and variants compared with those observed in the 1000 Genomes, single nucleotide polymorphism database (dbSNP) and HapMap databases. Functional effects of non-synonymous variants were predicted using Polyphen-2 and sorting intolerant from tolerant (SIFT).No disease-causing mutations were identified in NLRP2, NLRP7 and KHDC3L in our cohorts of unexplained infertility and RPL.Due to the limited patient size, it is difficult to conclude if the low frequency single nucleotide polymorphisms observed in the present study are causative of the phenotype. The design of the present study therefore is only capable of detecting highly penetrant mutations.The present study supports the hypothesis that mutations in NLRP7 and KHDC3L are specific for the BiHM phenotype and do not play a role in other adverse reproductive outcomes. Furthermore, to date, mutations in NLRP2 have only been associated with the imprinting disorder BWS in offspring and there is no evidence for a role in molar pregnancies, RPL or unexplained infertility.This study was funded by the following sources: Estonian Ministry of Education and Research (Grant SF0180044s09), Enterprise Estonia (Grant EU30020); Mentored Resident research project (Department of Obstetrics and Gynecology, Baylor College of Medicine); Imperial NIHR Biomedical Research Centre; Grant Number C06RR029965 from the National Center for Research Resources (NCCR; NIH). No competing interests declared.
View details for DOI 10.1093/humrep/deu296
View details for Web of Science ID 000350146100028
View details for PubMedID 25376457
View details for PubMedCentralID PMC4262469
Infiltrative Neurosarcoidosis Presenting as Secondary Amenorrhea Case Report and Review of the Literature
OBSTETRICAL & GYNECOLOGICAL SURVEY
2013; 68 (6): 482-488
In this report, we describe abrupt onset of secondary amenorrhea in a woman with history of chronic systemic sarcoidosis. Endocrinologic evaluation of her hypothalamic-pituitary axis revealed abnormally low levels of follicle-stimulating hormone, luteinizing hormone, and insulinlike growth factor 1 and elevated prolactin. Urine osmolality was low, and serum osmolality was high. Magnetic resonance imaging revealed diffuse extensive leptomeningeal enhancement, with involvement of the hypothalamus, pituitary stalk, and the optic chiasm. Clinical diagnosis was consistent with neurosarcoidosis with hypothalamic-pituitary infiltration resulting in clinical hypogonadotropic hypogonadism, hyper-prolactinemia, and diabetes insipidus. In our report, we provide an overview of basic reproductive neuroendocrinology and discuss salient concepts of the pathogenesis, clinical manifestations, evaluation, and management of hypogonadotropic hypogonadism. The current literature on neurosarcoidosis with involvement of the hypothalamic-pituitary axis is summarized. The possibility of infiltrative process should be considered in patients with new diagnosis of hypogonadotropic hypogonadal amenorrhea.
View details for DOI 10.1097/OGX.0b013e31828e116e
View details for Web of Science ID 000319559100021
View details for PubMedID 23942474
Perivascular Human Endometrial Mesenchymal Stem Cells Express Pathways Relevant to Self-Renewal, Lineage Specification, and Functional Phenotype
BIOLOGY OF REPRODUCTION
2012; 86 (2)
Human endometrium regenerates on a cyclic basis from candidate stem/progenitors whose genetic programs are yet to be determined. A subpopulation of endometrial stromal cells, displaying key properties of mesenchymal stem cells (MSCs), has been characterized. The endometrial MSC (eMSC) is likely the precursor of the endometrial stromal fibroblast. The goal of this study was to determine the transcriptome and signaling pathways in the eMSC to understand its functional phenotype. Endometrial stromal cells from oocyte donors (n = 20) and patients undergoing benign gynecologic surgery (n = 7) were fluorescence-activated cell sorted into MCAM (CD146)(+)/PDGFRB(+) (eMSC), MCAM (CD146)(-)/PDGFRB(+) (fibroblast), and MCAM (CD146)(+)/PDGFRB(-) (endothelial) populations. The eMSC population contained clonogenic cells with a mesenchymal phenotype differentiating into adipocytes when cultured in adipogenic medium. Gene expression profiling using Affymetrix Human Gene 1.0 ST arrays revealed 762 and 1518 significantly differentially expressed genes in eMSCs vs. stromal fibroblasts and eMSCs vs. endothelial cells, respectively. By principal component and hierarchical clustering analyses, eMSCs clustered with fibroblasts and distinctly from endothelial cells. Endometrial MSCs expressed pericyte markers and were localized by immunofluorescence to the perivascular space of endometrial small vessels. Endometrial MSCs also expressed genes involved in angiogenesis/vasculogenesis, steroid hormone/hypoxia responses, inflammation, immunomodulation, cell communication, and proteolysis/inhibition, and exhibited increased Notch, TGFB, IGF, Hedgehog, and G-protein-coupled receptor signaling pathways, characteristic of adult tissue MSC self-renewal and multipotency. Overall, the data support the eMSC as a clonogenic, multipotent pericyte that displays pathways of self-renewal and lineage specification, the potential to respond to conditions during endometrial desquamation and regeneration, and a genetic program predictive of its differentiated lineage, the stromal fibroblast.
View details for DOI 10.1095/biolreprod.111.095885
View details for Web of Science ID 000301341500017
View details for PubMedID 22075475
View details for PubMedCentralID PMC3290674
Nuclear Receptor, Coregulator Signaling, and Chromatin Remodeling Pathways Suggest Involvement of the Epigenome in the Steroid Hormone Response of Endometrium and Abnormalities in Endometriosis
2012; 19 (2): 152-162
Human endometrium, a steroid hormone-dependent tissue, displays complex cellular regulation mediated by nuclear receptors (NRs). The NRs interact with histone-modifying and DNA-methylating/-demethylating enzymes in the transcriptional complex. We investigated NRs, their coregulators, and associated signaling pathways in endometrium across the normal menstrual cycle and in endometriosis, an estrogen-dependent, progesterone-resistant disorder. Endometrial tissue was processed for analysis of 84 genes using NR and coregulator polymerase chain reaction (PCR) arrays. Select genes were validated by immunohistochemistry. Ingenuity pathway analysis identified DNA methylation and transcriptional repression signaling as the most affected pathway in endometrium in women with versus without endometriosis, regardless of cycle phase. Thyroid hormone receptor (THR) and vitamin D receptor (VDR) pathways were also regulated in normal and disease endometrium by activation of TH or vitamin D regulated genes. These data support the involvement of the epigenome in steroid hormone response of normal endometrium throughout the cycle and abnormalities in endometrium in women with endometriosis.
View details for DOI 10.1177/1933719111415546
View details for Web of Science ID 000300991100004
View details for PubMedID 22138541
View details for PubMedCentralID PMC3343132
- Sex selection for nonhealth-related reasons. The virtual mentor : VM 2012; 14 (2): 105-111
Comparative Transcriptome Analysis of Human Trophectoderm and Embryonic Stem Cell-Derived Trophoblasts Reveal Key Participants in Early Implantation
BIOLOGY OF REPRODUCTION
2012; 86 (1)
The implantation process begins with attachment of the trophectoderm (TE) of the blastocyst to the maternal endometrial epithelium. Herein we have investigated the transcriptome of mural TE cells from 13 human blastocysts and compared these with those of human embryonic stem cell (hESC)-derived-TE (hESC(troph)). The transcriptomes of hESC(troph) at Days 8, 10, and 12 had the greatest consistency with TE. Among genes coding for secreted proteins of the TE of human blastocysts and of hESC(troph) are several molecules known to be involved in the implantation process, as well as novel ones, such as CXCL12, HBEGF, inhibin A, DKK3, WNT5A, and follistatin. The similarities between the two lineages underscore some of the known mechanisms and offer discovery of new mechanisms and players in the process of the very early stages of human implantation. We propose that the hESC(troph) is a viable functional model of human trophoblasts to study trophoblast-endometrial interactions. Furthermore, the data derived herein offer the promise of novel diagnostics and therapeutics aimed at practical challenges in human infertility and pregnancy disorders associated with abnormal embryonic implantation.
View details for DOI 10.1095/biolreprod.111.092775
View details for Web of Science ID 000300548400010
View details for PubMedID 21865555
Biobanking human endometrial tissue and blood specimens: standard operating procedure and importance to reproductive biology research and diagnostic development
FERTILITY AND STERILITY
2011; 95 (6): 2120-U302
To develop a standard operating procedure (SOP) for collection, transport, storage of human endometrial tissue and blood samples, subject and specimen annotation, and establishing sample priorities.The SOP synthesizes sound scientific procedures, the literature on ischemia research, sample collection and gene expression profiling, good laboratory practices, and the authors' experience of workflow and sample quality.The National Institutes of Health, University of California, San Francisco, Human Endometrial Tissue and DNA Bank.Women undergoing endometrial biopsy or hysterectomy for nonmalignant indications.Collecting, processing, storing, distributing endometrial tissue and blood samples under approved institutional review board protocols and written informed consent from participating subjects.Standard operating procedure.The SOP addresses rigorous and consistent subject annotation, specimen processing and characterization, strict regulatory compliance, and a reference for researchers to track collection and storage times that may influence their research.The comprehensive and systematic approach to the procurement of human blood and endometrial tissue in this SOP ensures the high quality, reliability, and scientific usefulness of biospecimens made available to investigators by the National Institutes of Health, University of California, San Francisco, Human Endometrial Tissue and DNA Bank. The detail and perspective in this SOP also provides a blueprint for implementation of similar collection programs at other institutions.
View details for DOI 10.1016/j.fertnstert.2011.01.164
View details for Web of Science ID 000289620900053
View details for PubMedID 21371706
View details for PubMedCentralID PMC3080464
Unique Transcriptome, Pathways, and Networks in the Human Endometrial Fibroblast Response to Progesterone in Endometriosis
BIOLOGY OF REPRODUCTION
2011; 84 (4): 801-815
Eutopic endometrium in endometriosis has molecular evidence of resistance to progesterone (P(4)) and activation of the PKA pathway in the stromal compartment. To investigate global and temporal responses of eutopic endometrium to P(4), we compared early (6-h), intermediate (48-h), and late (14-Day) transcriptomes, signaling pathways, and networks of human endometrial stromal fibroblasts (hESF) from women with endometriosis (hESF(endo)) with hESF from women without endometriosis (hESF(nonendo)). Endometrial biopsy samples were obtained from subjects with and without mild peritoneal endometriosis (n = 4 per group), and hESF were isolated and treated with P(4) (1 μM) plus estradiol (E(2)) (10 nM), E(2) alone (10 nM), or vehicle for up to 14 days. Total RNA was subjected to microarray analysis using a Gene 1.0 ST (Affymetrix) platform and analyzed by using bioinformatic algorithms, and data were validated by quantitative real-time PCR and ELISA. Results revealed unique kinetic expression of specific genes and unique pathways, distinct biological and molecular processes, and signaling pathways and networks during the early, intermediate, and late responses to P(4) in both hESF(nonendo) and hESF(endo), although a blunted response to P(4) was observed in the latter. The normal response of hESF to P(4) involves a tightly regulated kinetic cascade involving key components in the P(4) receptor and MAPK signaling pathways that results in inhibition of E(2)-mediated proliferation and eventual differentiation to the decidual phenotype, but this was not established in the hESF(endo) early response to P(4). The abnormal response of this cell type to P(4) may contribute to compromised embryonic implantation and infertility in women with endometriosis.
View details for DOI 10.1095/biolreprod.110.086181
View details for Web of Science ID 000288596900023
View details for PubMedID 20864642
View details for PubMedCentralID PMC3062042
Effect of bisphenol A on human endometrial stromal fibroblasts in vitro
REPRODUCTIVE BIOMEDICINE ONLINE
2011; 22 (3): 249-256
This study evaluated the effects of bisphenol A (BPA) on human endometrial stromal fibroblast (ESF) differentiation and expression of genes involved in oestrogen metabolism. Human ESF from eight hysterectomy specimens were cultured and treated with 5-100 μmol/l of BPA ± oestradiol or 8-br-cAMP for 48 h. mRNA expression was analysed by real-time reverse-transcription PCR. 8-br-cAMP-induced human ESF decidualization was confirmed by expression of insulin-like growth factor binding protein-1 (IGFBP1) and prolactin secretion. Short-term exposure (48 h) decreased human ESF proliferation (P<0.04) not due to apoptosis. High doses of BPA significantly induced IGFBP1 mRNA and protein, decreased P450scc mRNA, reversed the 8-br-cAMP-induced increase in HSD17B2 (oestradiol to oestrone conversion) in a dose-dependent manner and down-regulated HSD17B1 expression (oestrone to oestradiol conversion; P ≤ 0.03). 8-br-cAMP significantly potentiated this effect (P=0.028). BPA had no significant effect on aromatase and PPAR γ expression. The oestrogen-receptor antagonist ICI had no effect on gene expression in BPA-treated cells, and oestrogen receptor α, but not oestrogen receptor β, was significantly down-regulated by high doses of BPA (P=0.028). BPA has an endocrine-disrupting effect on human ESF function and gene expression but the underlying mechanisms appear not to involve oestrogen-mediated pathways.
View details for DOI 10.1016/j.rbmo.2010.12.007
View details for Web of Science ID 000303040900003
View details for PubMedID 21273127
View details for PubMedCentralID PMC3836676
Molecular Evidence for Differences in Endometrium in Severe Versus Mild Endometriosis
2011; 18 (3): 229-251
Women with stage III/IV versus stage I/II endometriosis have lower implantation and pregnancy rates in natural and assisted reproduction cycles. To elucidate potential molecular mechanisms underlying these clinical observations, herein we investigated the transcriptome of eutopic endometrium across the menstrual cycle in the setting of severe versus mild endometriosis. Proliferative (PE), early secretory (ESE), and mid-secretory (MSE) endometrial tissues were obtained from 63 participants with endometriosis (19 mild and 44 severe). Purified RNA was subjected to microarray analysis using the Gene 1.0 ST Affymetrix platform. Data were analyzed with GeneSpring and Ingenuity Pathway Analysis and subsequently validated. Comparison of differentially regulated genes, analyzed by cycle phase, revealed dysregulation of progesterone and/or cyclic adenosine monophosphate (cAMP)-regulated genes and genes related to thyroid hormone action and metabolism. Also, members of the epidermal growth factor receptor (EGFR) signaling pathway were observed, with the greatest upregulation of EGFR in severe versus mild disease during the early secretory phase. The extracellular matrix proteoglycan versican (VCAN), which regulates cell proliferation and apoptosis, was the most highly expressed gene in severe versus mild disease. Upregulation of microRNA 21 (MIR21) and DICER1 transcripts suggests roles for microRNAs (miRNAs) in the pathogenesis of severe versus mild endometriosis, potentially through regulation of gene silencing and epigenetic mechanisms. These observed differences in transcriptomic signatures and signaling pathways may result in poorly programmed endometrium during the cycle, contributing to lower implantation and pregnancy rates in women with severe versus mild endometriosis.
View details for DOI 10.1177/1933719110386241
View details for Web of Science ID 000290164400003
View details for PubMedID 21063030
View details for PubMedCentralID PMC3118406
Thyroid-stimulating hormone receptor and thyroid hormone receptors are involved in human endometrial physiology
FERTILITY AND STERILITY
2011; 95 (1): 230-U798
To study the expression, distribution, and function of thyroid-stimulating hormone receptor (TSHR) and thyroid hormone receptors (TR) α1, α2, and β1 in human endometrium.Experimental clinical study.University hospital.31 fertile women.Endometrial biopsy samples obtained throughout the menstrual cycle.Real-time reverse transcriptase polymerase chain reaction, immunohistochemistry and Western blot to study the expression of TSHR, TRα1, TRα2, and TRβ1 messenger RNA (mRNA) and proteins in human endometrium.We found TSHR, TRα1, TRα2 and TRβ1 mRNA and proteins expressed in human endometrium. Immunostaining for TSHR in the luminal epithelium and TRα1 and β1 in the glandular and luminal epithelium increased statistically significantly on luteinizing hormone (LH) days 6 to 9, coinciding with appearance of pinopodes. Endometrial stromal and Ishikawa cells expressed mRNA for TSHR, TR, and iodothyronine deiodinases 1-3. After 48 hours, TSH significantly increased leukemia inhibitory factor (LIF) and LIF receptor (LIFR) messenger RNA (mRNA) in endometrial stromal cells, but decreased their expression in Ishikawa cells. Glucose transporter 1 mRNA was up-regulated by TSH in Ishikawa cells. We found that TSH statistically significantly increased secretion of free triiodothyronine (T3) and total thyroxin (T4) by Ishikawa cells compared with nonstimulated cells.Thyroid hormones are directly involved in endometrial physiology.
View details for DOI 10.1016/j.fertnstert.2010.06.079
View details for Web of Science ID 000285411600047
View details for PubMedID 20691434
Diminished Endometrial Expression of Ghrelin and Ghrelin Receptor Contributes to Infertility
2010; 17 (9): 823-832
The objectives were to investigate the presence, distribution and sex steroid hormone regulation of ghrelin and its receptor, growth hormone secretagogue receptor (GHSR), in human endometrium in relation to endometrial receptivity and fertility. Endometrial biopsies were obtained from women with unexplained infertility and healthy fertile volunteers. Ishikawa cells were used to mimic the action of ghrelin in endometrium. Immunostaining of GHSR was strong in luminal epithelium and stroma during mid-secretory phase. Ghrelin and GHSR expression is less intense in mid-secretory endometrium of infertile women compared to fertile controls. Treatment with estrogen and/or progesterone or their antagonists did not significantly change the relative expression of GHSR in Ishikawa and stromal cells. Ghrelin was present in and secreted from human blastocysts, which suggest that the communication between human blastocyst and endometrium might involve ghrelin. Low levels of GHSR in endometrium from women with unexplained infertility may in part explain the infertility.
View details for DOI 10.1177/1933719110371683
View details for Web of Science ID 000282003400005
View details for PubMedID 20616368
The Bone Marrow-Derived Human Mesenchymal Stem Cell: Potential Progenitor of the Endometrial Stromal Fibroblast
BIOLOGY OF REPRODUCTION
2010; 82 (6): 1076-1087
The cellular sources that contribute to the renewal of human endometrium are largely unknown. It has been suggested that endometrial stem cells originate from bone marrow-derived mesenchymal stem cells (MSC), with subsequent development into endometrial stromal fibroblasts (hESF). We hypothesized that if bone marrow-derived MSC contribute to endometrial regeneration and are progenitors of hESF, their treatment with agents known to regulate hESF differentiation could promote their differentiation down the stromal fibroblast lineage. To this end, we treated bone marrow-derived MSC with estradiol and progesterone, bone morphogenetic protein 2 (BMP2), and activators of the protein kinase A (PKA) pathway and investigated specific markers of hESF differentiation (decidualization). Furthermore, we investigated the transcriptome of these cells in response to cAMP and compared this to the transcriptome of hESF decidualized in response to activation of the PKA pathway. The data support the idea that MSC can be differentiated down the hESF pathway, as evidenced by changes in cell shape and common expression of decidual markers and other genes important in hESF differentiation and function, and that bone marrow-derived MSC may be a source of endometrial stem/progenitor cells. In addition, we identified MSC-specific markers that distinguish them from other fibroblasts and, in particular, from hESF, which is of biologic relevance and practical value to the field of endometrial stem cell research.
View details for DOI 10.1095/biolreprod.109.082867
View details for Web of Science ID 000277964500008
View details for PubMedID 20147733
View details for PubMedCentralID PMC2874495
Update on the role of leukemia inhibitory factor in assisted reproduction
CURRENT OPINION IN OBSTETRICS & GYNECOLOGY
2010; 22 (3): 213-219
To review the recent literature on the involvement and importance of leukemia inhibitory factor (LIF) in the human implantation process, and the attempts using LIF-based interventions to improve assisted reproductive technologies (ARTs) outcome in women with recurrent implantation failure.High LIF expression is an indicator of receptive endometrium in fertile women. However, in infertile individuals, the data on endometrial LIF expression and secretion are controversial. Even after ruling out other causes of infertility, such as tubal, endocrine, male factor, and endometriosis, LIF-only detection is not sufficient for assessment of implantation potential in women with unexplained infertility. This is obviously in contrast to evidence of the crucial role of LIF in mouse endometrial physiology. In a large multicenter study, recombinant human LIF failed to improve the outcome of IVF treatment in women with recurrent implantation failure.A better comprehension of the mechanisms underlying endometrial receptivity and implantation should guide clinicians through proper management and treatment of infertility and implantation failure, and may eventually enable widespread adherence to single embryo transfer practices.
View details for DOI 10.1097/GCO.0b013e32833848e5
View details for Web of Science ID 000278201000007
View details for PubMedID 20216416
The Protein Kinase A Pathway-Regulated Transcriptome of Endometrial Stromal Fibroblasts Reveals Compromised Differentiation and Persistent Proliferative Potential in Endometriosis
2010; 151 (3): 1341-1355
Intrinsic abnormalities in transplanted eutopic endometrium are believed to contribute to the pathogenesis of pelvic endometriosis. Herein we investigated transcriptomic differences in human endometrial stromal fibroblasts (hESFs) from women with (hESF(endo)) vs. without (hESF(nonendo)) endometriosis, in response to activation of the protein kinase A (PKA) pathway with 8-bromoadenosine-cAMP (8-Br-cAMP). hESF(nonendo) (n = 4) and hESF(endo) (n = 4) were isolated from eutopic endometrium and treated +/- 0.5 mm 8-Br-cAMP for 96 h. Purified total RNA was subjected to microarray analysis using the whole-genome Gene 1.0 ST Affymetrix platform. A total of 691 genes were regulated in cAMP-treated hESF(nonendo) vs. 158 genes in hESF(endo), suggesting a blunted response to cAMP/PKA pathway activation in women with disease. Real-time PCR and ELISA validated the decreased expression of decidualization markers in hESF(endo) compared with hESF(nonendo). In the absence of disease, 8-Br-cAMP down-regulated progression through the cell cycle via a decrease in cyclin D1, cyclin-dependent kinase 6, and cell division cycle 2 and an increase in cyclin-dependent kinase inhibitor 1A. However, cell cycle components in hESF(endo) were not responsive to 8-Br-cAMP, resulting in persistence of a proliferative phenotype. hESF(endo) treated with 8-Br-cAMP exhibited altered expression of immune response, extracellular matrix, cytoskeleton, and apoptosis genes. Changes in phosphodiesterase expression and activity were not different among experimental groups. These data support that eutopic hESF(endo) with increased proliferative potential can seed the pelvic cavity via retrograde menstruation and promote establishment, survival, and proliferation of endometriosis lesions, independent of hydrolysis of cAMP and likely due to an inherent abnormality in the PKA pathway.
View details for DOI 10.1210/en.2009-0923
View details for Web of Science ID 000274711600054
View details for PubMedID 20068008
View details for PubMedCentralID PMC2840687
Altered Gene Expression Profiling in Endometrium: Evidence for Progesterone Resistance
SEMINARS IN REPRODUCTIVE MEDICINE
2010; 28 (1): 51-58
Progesterone plays an important role in regulating multiple events in the uterus. It controls endometrial proliferation and differentiation, which are important for uterine function. Dysregulation of progesterone signaling leads to impaired physiological functions. Indeed, aberrant expression of progesterone-regulated genes in the endometrium has been implicated in several gynecologic disorders, including endometriosis, polycystic ovarian syndrome (PCOS), and endometrial hyperplasia. Although several investigators have analyzed eutopic endometrial expression of progesterone-target genes, the genesis and consequences of progesterone resistance remain unclear. We review evidence for progesterone resistance in endometrium of women with endometriosis, PCOS, and endometrial hyperplasia, and we identify possible mechanisms associated with reduced progesterone activity in endometrium of (some) women with these gynecologic disorders that have a significant impact on women's health and well-being.
View details for DOI 10.1055/s-0029-1242994
View details for Web of Science ID 000273921100007
View details for PubMedID 20104428
Increased Mitogen-Activated Protein Kinase Kinase/Extracellularly Regulated Kinase Activity in Human Endometrial Stromal Fibroblasts of Women with Endometriosis Reduces 3 ',5 '-Cyclic Adenosine 5 '-Monophosphate Inhibition of Cyclin D1
2009; 150 (10): 4701-4712
Endometriosis is characterized by endometrial tissue growth outside the uterus, due primarily to survival, proliferation, and neoangiogenesis of eutopic endometrial cells and fragments refluxed into the peritoneal cavity during menses. Although various signaling molecules, including cAMP, regulate endometrial proliferation, survival, and embryonic receptivity in endometrium of women without endometriosis, the exact molecular signaling pathways in endometrium of women with disease remain unclear. Given the persistence of a proliferative profile and differential expression of genes associated with the MAPK signaling cascade in early secretory endometrium of women with endometriosis, we hypothesized that ERK1/2 activity influences cAMP regulation of the cell cycle. Here, we demonstrate that 8-Br-cAMP inhibits bromodeoxyuridine incorporation and cyclin D1 (CCND1) expression in cultured human endometrial stromal fibroblasts (hESF) from women without but not with endometriosis. Incubation with serum-containing or serum-free medium resulted in higher phospho-ERK1/2 levels in hESF of women with vs. without disease, independent of 8-Br-cAMP treatment. The MAPK kinase-1/2 inhibitor, U0126, fully restored cAMP down-regulation of CCND1, but not cAMP up-regulation of IGFBP1, in hESF of women with vs. without endometriosis. Immunohistochemistry demonstrated the highest phospho-ERK1/2 in the late-secretory epithelial and stromal cells in women without disease, in contrast to intense immunostaining in early-secretory epithelial and stromal cells in those with disease. These findings suggest that increased activation of ERK1/2 in endometrial cells from women with endometriosis may be responsible for persistent proliferative changes in secretory-phase endometrium.
View details for DOI 10.1210/en.2009-0389
View details for Web of Science ID 000270021700025
View details for PubMedID 19589865
View details for PubMedCentralID PMC2754675
MicroRNA expression profiling of eutopic secretory endometrium in women with versus without endometriosis
MOLECULAR HUMAN REPRODUCTION
2009; 15 (10): 625-631
Endometriosis is a common gynecologic disorder characterized by pain and infertility. In addition to estrogen dependence, progesterone resistance is an emerging feature of this disorder. Specifically, a delayed transition from the proliferative to secretory phase as evidenced by dysregulation of progesterone target genes and maintenance of a proliferative molecular fingerprint in the early secretory endometrium (ESE) has been reported. MicroRNAs (miRNAs) are small noncoding RNAs that collectively represent a novel class of regulators of gene expression. In an effort to investigate further the observed progesterone resistance in the ESE of women with endometriosis, we conducted array-based, global miRNA profiling. We report distinct miRNA expression profiles in the ESE of women with versus without endometriosis in a subset of samples previously used in global gene expression analysis. Specifically, the miR-9 and miR-34 miRNA families evidenced dysregulation. Integration of the miRNA and gene expression profiles provides unique insights into the molecular basis of this enigmatic disorder and, possibly, the regulation of the proliferative phenotype during the early secretory phase of the menstrual cycle in affected women.
View details for DOI 10.1093/molehr/gap068
View details for Web of Science ID 000270005200005
View details for PubMedID 19692421
View details for PubMedCentralID PMC2744474
The Progesterone Receptor Coactivator Hic-5 Is Involved in the Pathophysiology of Endometriosis
2009; 150 (8): 3863-3870
Endometriosis is an estrogen-dependent disorder primarily associated with pelvic pain and infertility in up to 10% of women of reproductive age. Recent studies suggest that resistance to progesterone action may contribute to the development and pathophysiology of this disorder. In this study we examined the in vivo and in vitro expression and function of one progesterone receptor (PR) coactivator, Hic-5, in human endometrium and endometrial stromal fibroblasts (hESFs) from 29 women with and 30 (control) women without endometriosis. Hic-5 was highly expressed in stromal, but not epithelial, cells in women without endometriosis, in a cycle-dependent manner. In contrast, Hic-5 expression was not regulated during the menstrual cycle in hESFs from women with endometriosis and was significantly reduced in hESFs from women with vs. without disease. Hic-5 mRNA expression throughout the cycle in endometrium from control women, but not those with endometriosis, correlated with expression of PR. Hic-5 mRNA in hESFs was significantly up-regulated in control but not endometriosis hESFs after treatment in vitro with 8-bromoadenosine-cAMP for 96 h but only modestly after 14 d of progesterone treatment. Hic-5 silencing did not influence cAMP-regulated gene expression but affected genes regulated solely by progesterone (e.g. DKK1 and calcitonin). Together the data suggest that the proposed progesterone resistance in endometrium from women with endometriosis derives, in part, from impaired expression of the PR coactivator, Hic-5, in endometrial tissue and cultured endometrial stromal fibroblasts.
View details for DOI 10.1210/en.2009-0008
View details for Web of Science ID 000268158400052
View details for PubMedID 19389829
View details for PubMedCentralID PMC2717860
Disturbances in the LIF pathway in the endometrium among women with unexplained infertility
FERTILITY AND STERILITY
2009; 91 (6): 2602-2610
To study the expression of leukemia inhibitory factor (LIF), its receptors LIFR and gp130, and its inhibitor SOCS1 in endometria from fertile women and infertile women with unexplained infertility. Signaling through the LIF pathway is involved in maintenance of a receptive state of human endometrium. Impaired endometrial receptivity may be a cause of female infertility.Prospective clinical study.Hospital-based IVF unit and university-affiliated reproductive research laboratories.Twenty-six healthy fertile women and 14 women with unexplained infertility.Endometrial biopsy.Pinopode formation, expression of LIF, LIFR, gp130, and SOCS1 protein and mRNA in endometrial biopsies.The expression of LIFR in the endometrium was negatively correlated to the expression of SOCS1 and positively correlated to the formation of pinopodes. In control fertile women, simultaneous intense apical staining of LIFR and gp130 together with faint SOCS1 staining was observed in epithelial cells, whereas the opposite was seen in most women with unexplained infertility.Unexplained infertility in some women might be explained by disturbances in the LIF pathway in midsecretory-phase endometrium.
View details for DOI 10.1016/j.fertnstert.2008.04.010
View details for Web of Science ID 000266801400049
View details for PubMedID 18684446
Receptors for thyroid-stimulating hormone and thyroid hormones in human ovarian tissue
REPRODUCTIVE BIOMEDICINE ONLINE
2009; 18 (3): 337-347
Dysfunction in thyroid regulation can cause menstrual and ovulatory disturbances, the mechanism of which is not clear. The distribution and activity of the thyroid-stimulating hormone (TSHR), and the thyroid hormone receptors (TR) alpha1, alpha2 and beta1 in human ovarian tissue and in granulosa cells was studied using immunohistochemistry, reverse-transcriptase polymerase chain reaction (RT-PCR), quantitative PCR and immunoassays. Strong immunostaining of TSHR, TRalpha1 and TRbeta1 was observed in ovarian surface epithelium and in oocytes of primordial, primary and secondary follicles, with minimal staining in granulosa cells of secondary follicles. Granulosa cells of antral follicles expressed TSHR, TRalpha1 and TRbeta1 proteins. Messenger RNA for all receptors was present in ovarian tissue. Mature human granulosa cells expressed transcripts for 5' deiodinases types 2 and 3, but not type 1, indicating the possibility of conversion of peripheral thyroid hormone thyroxin (T(4)). Granulosa cells stimulated with TSH showed a significant increase in cAMP concentrations after 2 h of culture (P = 0.047), indicating activation through TSHR. Stimulation with T(4) resulted in increased extracellular signal-regulated kinase 1 and 2 activation after 10, 30, 60 min and 24 h. These data demonstrate that TSH and thyroid hormone receptors may participate in the regulation of ovarian function.
View details for Web of Science ID 000264455200005
View details for PubMedID 19298732
Steroidogenic Enzyme and Key Decidualization Marker Dysregulation in Endometrial Stromal Cells from Women with Versus Without Endometriosis
BIOLOGY OF REPRODUCTION
2009; 80 (1): 105-114
Identification of mechanisms underlying endometriosis pathogenesis will facilitate understanding and treatment of infertility and pain associated with this disorder. Herein, we investigated the expression of steroidogenic pathway enzymes and key decidualization biomarkers in endometrial tissue and in eutopic endometrial stromal fibroblasts (hESFs) from women with vs. those without endometriosis, and subsequently treated in vitro with 8-bromo-cAMP (8-Br-cAMP) or progesterone (P4). Real-time quantitative PCR, immunohistochemistry, ELISA, and radiometric aromatase activity assay were used. The results demonstrate significantly increased (14.5-fold; P=0.037) expression of aromatase in eutopic endometrium of women with disease. In 8-Br-cAMP-treated hESF from eutopic endometrium of women with endometriosis, the balance in estradiol (E2) and P4 biosynthetic and metabolizing enzymes is disturbed (decreased HSD3B1 and HSD17B2, and increased HSD17B1 and aromatase), with the equilibrium being shifted towards an E2-enriched milieu. However, hESF from the same group of women treated with P4 did not demonstrate such responsiveness. Lower expression of IGFBP1 and prolactin mRNA and protein was observed in hESF from women with vs. those without endometriosis in response to 8-Br-cAMP, but not P4, suggesting a blunted response of these decidual biomarkers to activation of the PKA pathway in eutopic endometrium in women with disease. The dichotomy of 8-Br-cAMP regulation of select steroidogenic enzymes leading to an enriched E2 milieu within the endometrium and a blunted response of decidual biomarkers to this decidualizing agent of hESF from women with endometriosis suggests resistance to full decidualization of the stromal fibroblasts and mechanisms underlying implantation failure and the pathophysiology of this disorder.
View details for DOI 10.1095/biolreprod.108.070300
View details for Web of Science ID 000262010100012
View details for PubMedID 18815356
View details for PubMedCentralID PMC2704986
HB-EGF but not amphiregulin or their receptors HER1 and HER4 is altered in endometrium of women with unexplained infertility
2008; 15 (5): 484-492
Heparin-binding epidermal growth factor-like growth factor (HB-EGF) and its receptors (HER1 and HER4) play a role in the human implantation process. Amphiregulin is a member of the EGF family but with unknown function in human fertility. It has been suggested that some women with unexplained infertility have defective endometrial development. The aim of this study is to determine the presence of amphiregulin and the receptors HER1 and HER4 in normal human endometrium throughout the menstrual cycle. In addition, the present study aims to compare endometrium from women with unexplained infertility with endometrium from women with male factor infertility and healthy fertile controls. Immunohistochemistry and real-time polymerase chain reaction were used to determine the expression of HB-EGF, HER1, HER4, and amphiregulin. The stromal staining of HER1 and the epithelial staining of HER4 were most intense in the mid- and late-secretory-phase endometrium. Amphiregulin did not vary during the menstrual cycle. In the mid-secretory phase, the protein expression of HB-EGF was lower in endometrium from women with unexplained infertility versus normal endometrium and endometrium from women with male factor infertility. HB-EGF and HER4 mRNA expression in mid-secretory endometrium of women with unexplained and male factor infertility were increased compared with normal controls. Impaired endometrial expression of certain members of the EGF family may contribute to infertility in some women with unexplained infertility.
View details for DOI 10.1177/19337191083t4624
View details for Web of Science ID 000257086700009
View details for PubMedID 18579857
Uterine receptivity to human embryonic implantation: Histology, biomarkers, and transcriptomics
SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY
2008; 19 (2): 204-211
Embryonic implantation is a dynamic process of paracrine interactions between the maternal compartment and the conceptus and involves a receptive endometrium and a developmentally competent blastocyst. Herein, we review histology, clinical approaches, and the promise of transcriptomics in elucidating mechanisms underlying implantation and development of biomarkers of uterine receptivity-with an eye to diagnose and treat implantation-based disorders of miscarriage, fetal growth restriction, pre-eclampsia, and infertility.
View details for DOI 10.1016/j.semcdb.2007.10.008
View details for Web of Science ID 000255730300016
View details for PubMedID 18035563
View details for PubMedCentralID PMC2829661
Glycodelin is present in pinopodes of receptive-phase human endometrium and is associated with down-regulation of progesterone receptor B
FERTILITY AND STERILITY
2006; 85 (6): 1803-1811
To test the hypothesis that glycodelin is localized on pinopodes and correlates with temporal immunostaining of leukemia inhibitory factor (LIF), LIF receptor (LIFR), and progesterone receptor B (PRB).Prospective clinical study.Hospital-based reproductive health unit and research laboratories.Twenty-five healthy fertile women with normal menstrual cycles.Endometrial biopsy specimens were obtained from healthy fertile women in the luteal phase of the menstrual cycle.Immunohistochemical staining of glycodelin, ultrastructural immunostaining of glycodelin, and double staining of glycodelin and PRB.Glycodelin is present in the glands when pinopodes appear. Glycodelin is localized on pinopodes but is also secreted from luminal epithelial cells regardless of pinopode formation. There was a negative correlation between glycodelin secretion from the glands and PRB staining. A weak correlation between the presence of LIFR (but not LIF) and glycodelin was found.Pinopode appearance, intense staining of LIFR in pinopodes and glycodelin staining in the glands are synchronized events. Down-regulation of PRB in the endometrium is concomitant with the presence of glycodelin in the endometrium, suggesting interaction.
View details for DOI 10.1016/j.fertnstert.2005.12.018
View details for Web of Science ID 000238427700029
View details for PubMedID 16759928
Expression of leukemia inhibitory factor and its receptors is increased during differentiation of human embryonic stem cells.
Fertility and sterility
2006; 86 (4 Suppl): 1193–1209
To investigate gene expression profiles during the early spontaneous differentiation of human embryonic stem cells (hESCs), with particular emphasis on leukemia inhibitory factor (LIF)-induced pathways and the ultrastructural surface morphology of the undifferentiated and spontaneously differentiated hESCs.Prospective experimental study.University laboratory.Four hESC cell lines.The effect of LIF on receptor expression level was studied in cultures.Gene expression in the hESC line HS237 was analyzed using microarrays. Real-time reverse-transcription polymerase chain reaction was used to validate the microarray results in four hESC lines (HS181, HS235, HS237, HS293). Immunohistochemistry was used to assay LIF, LIF receptor, and gp130 protein expression. Cell surface morphology was studied using scanning electron microscopy.The expression of LIF, LIF receptor, and gp130 messenger RNA and protein was increased in spontaneously differentiated HS237 cells compared with undifferentiated cells, with high expression of an inhibitor of LIF-mediated signaling, suppressor of cytokine signaling-1, in undifferentiated hESCs. Genes, those expressed specifically and those shared in undifferentiated hESCs, differentiated cells, and in fibroblasts, were identified. Supplementation with LIF did not affect the LIF receptor expression.The expression of LIF and its receptors is low in undifferentiated hESCs but increases during differentiation. Added LIF does not prevent spontaneous differentiation. Suppressor of cytokine signaling-1 may prevent LIF signaling in hESCs.
View details for DOI 10.1016/j.fertnstert.2005.12.081
View details for PubMedID 16949591
Gender-specific alteration of adrenergic responses in small femoral arteries from estrogen receptor-beta knockout mice
2005; 46 (5): 1163-1168
Estrogen receptor-beta knockout mice become hypertensive as they age, and males have a higher blood pressure than females. We hypothesized that the absence of estrogen receptor-beta may contribute to development of cardiovascular dysfunction by modification of adrenergic responsiveness in the peripheral vasculature. Small femoral arteries (internal diameter <200 microm) were isolated from estrogen receptor-beta knockout and wild-type mice and mounted on a wire myograph. Concentration-response curves to phenylephrine and norepinephrine were compared and the contribution of adrenoceptor subtypes established using specific agonists and antagonists. The involvement of endothelial factors in the modulation of resting tone was also investigated and immunohistochemical analysis used to confirm the presence or absence of estrogen receptor expression. Compared with wild type, arteries from estrogen receptor-beta knockout male, but not female, mice demonstrated gender-specific enhancement of the response to phenylephrine (alpha1-adrenoceptor agonist), which was accompanied by elevated basal tension attributable to endothelial factors. Contractile responses to the mixed adrenoceptor agonist norepinephrine did not differ significantly between estrogen receptor-beta knockout and wild type; however, beta-adrenoceptor inhibition unmasked an enhanced underlying alpha1-adrenoceptor responsiveness in estrogen receptor-beta knockout males. beta-adrenoceptor-mediated dilatation was also enhanced in estrogen receptor-beta knockout versus wild-type males. We suggest that estrogen receptor-beta modifies the adrenergic control of small artery tone in males but not in females.
View details for DOI 10.1161/01.HYP.0000185648.48498.c1
View details for Web of Science ID 000233544900017
View details for PubMedID 16216990
Optimizing cryopreservation of human testicular tissue: comparison of protocols with glycerol, propanediol and dimethylsulphoxide as cryoprotectants
2005; 20 (6): 1676-1687
Cryopreservation of testicular tissue is an option in fertility preservation for pre-pubertal boys who will lose spermatogenic cells as a result of chemotherapy. We compared three different protocols and cryoprotectants in cryopreservation of testicular tissue.Testicular tissue obtained from 16 infertile men was evaluated by light microscopy(LM), immunostaining against MAGE-A4, transmission electron microscopy (TEM) and organ culture. Seminiferous tubules (1312) from non-frozen (n = 16) and frozen-thawed samples (n = 34) were studied following cryopreservation using protocols with either 1,2-propanediol (PrOH), glycerol or dimethylsulphoxide (DMSO) as cryoprotectants.Normal structure was seen in 86 +/- 6% (mean +/- SD) of the fresh tissue. After freezing with DMSO, 70 +/- 6% and after PrOH, 37+/-3% of the tubules were judged to be good. When glycerol was used, the structure of the basal compartment of the tubules was severely damaged. The ultrastructure of the cryopreserved samples as revealed by TEM and MAGE-positive spermatogonia confirmed the findings. Cryopreserved Leydig cells maintained their morphology and ability to release testosterone in culture.DMSO as a cryoprotectant (at a 0.7 mol/l concentration) proved to maintain the structure of testicular tissue, especially spermatogonia, after cryopreservation better than PrOH or glycerol.
View details for DOI 10.1093/humrep/deh797
View details for Web of Science ID 000229286200036
View details for PubMedID 15860503
Leukemia inhibitory factor and human embryo implantation
UTERUS AND HUMAN REPRODUCTION
2004; 1034: 176-183
The success of embryonic implantation relies on an ideal cross-talk between the embryo and the receptive endometrium. This article focuses on the role of leukemia inhibitory factor (LIF) and its receptors in human embryo implantation. LIF is a secreted glycoprotein first described as a factor that induced the differentiation of mouse myeloid leukemic M1 cells into macrophages and later proposed as a marker of the embryo implantation process. An important role for LIF in implantation was shown on LIF knockout mice, when embryo implantation did not occur. In endometrium of healthy women, LIF and LIF mRNA are expressed throughout the menstrual cycle with a striking increase in the midsecretory phase, coinciding with a supposed window of implantation. Correlation in the expression of LIF and some other markers of implantation has been reported. LIF acts on cells by binding to the LIF receptor (LIFR) and gp130. Human blastocysts express mRNAs for LIFR and gp130, participating actively in establishing contact with the endometrium. In the endometrium, LIFR and gp130 are expressed in the endometrial epithelium throughout the cycle with strong increase in the midsecretory phase. Endometrium of infertile women produces significantly less LIF during the period of receptivity. The role of LIF gene mutations in unexplained infertility and implantation failures in IVF patients is not clear yet. Infertile patients showed reduced secretion of LIFR and gp130 compared with fertile controls during the implantation window. Recombinant human LIF might help to improve the implantation rate in women with unexplained infertility.
View details for DOI 10.1196/annals.1335.020
View details for Web of Science ID 000228130100016
View details for PubMedID 15731310
Coexpression of pinopodes and leukemia inhibitory factor, as well as its receptor, in human endometrium
FERTILITY AND STERILITY
2003; 79: 808-814
To determine cell-type-specific expression of leukemia inhibitory factor (LIF) and LIF receptor (LIFR) proteins relative to formation of pinopodes in human endometrial samples.Prospective clinical study.Hospital-based unit for reproductive health and university-affiliated reproductive research laboratories.Twenty-six healthy fertile women with normal menstrual cycles.Routine blood and urine samples were obtained, and vaginal ultrasonography and endometrial biopsy were performed. Pinopode formation and expression of LIF and LIFR were examined in endometrial samples.Samples obtained during LH days 6 through 9 had pinopodes at different developmental stages. Both surface and glandular epithelial cells expressed maximal levels of LIF and LIFR protein, in biopsy samples showed fully developed pinopodes. Immunostaining of LIF was more intense in the glandular epithelium, whereas immunostaining of LIFR was most intense in the surface epithelium. Before and after the appearance of pinopodes, LIF and LIFR immunostaining was less intense or faint. Stromal endometrial cells showed faint LIF accumulation.The simultaneous positive spatial and temporal expression of pinopodes and LIF and LIFR proteins in endometrial samples from healthy women suggests that both molecular and structural cell changes are important in the initiation of human blastocyst implantation.
View details for DOI 10.1016/S0015-0282(02)04830-6
View details for Web of Science ID 000181670300022
View details for PubMedID 12620495
Co-existence of heparin-binding epidermal growth factorlike growth factor and pinopodes in human endometrium at the time of implantation
MOLECULAR HUMAN REPRODUCTION
2002; 8 (8): 765-769
Pinopodes have been suggested to be markers of uterine receptivity. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is expressed in increasing amounts in the secretory phase endometrium and is considered to be important for the human implantation process. The aim of this study was to investigate a possible co-existence of pinopodes and HB-EGF in the normal human endometrium. Endometrial biopsies were obtained from women with normal menstrual cycles. The biopsies were examined by scanning electron microscopy for the detection of pinopodes, by immunohistochemistry for the expression of HB-EGF protein, and by confocal microscopy to determine if HB-EGF was present on the surface of the pinopodes. The expression of HB-EGF in luminal and glandular epithelium was highest when fully developed pinopodes were present. Using confocal microscopy it was shown that HB-EGF was present both inside the luminal epithelial cells and on the surface of pinopodes. These findings suggest that HB-EGF might play a role in both the attachment and penetration steps in the human implantation process. Furthermore, the immunohistochemical staining demonstrates that HB-EGF can be used as a marker for the implantation window.
View details for Web of Science ID 000177470200011
View details for PubMedID 12149409
Endometrial pinopodes: some more understanding on human implantation?
Reproductive biomedicine online
2002; 4: 18-23
Endometrial receptivity is a prerequisite for blastocyst implantation. During receptivity, the hairy-like epithelial cell microvilli transiently fuse to a single flower-like membrane projection called the 'pinopode'. Scanning electron microscopy in sequential endometrial biopsies shows that pinopodes appear about 1 week after ovulation, and they develop and regress within just 2 days. Interestingly, the cycle days when pinopodes appear can vary by up to 5 days between different individuals. On average, they occur on days 20-21 in natural cycles and earlier (days 19-20) in stimulated cycles. The abundance of pinopodes relates to implantation success and many patients with multiple implantation failures fail to produce pinopodes. Based on these findings, biopsies from candidate embryo recipients have been examined in mock cycles and pinopode numbers and timing of their appearance assessed. A similar cycle follows where embryos are replaced earlier or later, according to the reported timing of pinopode formation. If pinopodes are absent, the cycle can be modified. Accumulating evidence supports their clinical use as a marker to assess endometrial receptivity. Pinopode appearance, loss of steroid receptors and maximal expression of a(v)b(3) integrin, osteopontin and leukaemia inhibitory factor and receptor have been demonstrated in the same biopsy, showing a consistent association of pinopode appearance and other receptivity changes.
View details for PubMedID 12470560