In humans, exposure to xenoestrogens such as diethylstilbestrol (DES) and bisphenol A (BPA) has been associated with developmental disorders of the female reproductive tract. The investigators have previously shown that fetal exposure to these agents alters the methylation of estrogen response elements (EREs) in the endometrium. The change in ERE methylation results in altered ERE occupancy by estrogen receptor (ER) in vivo and altered gene expression in response to estrogens throughout life. The uterine endometrium is replaced in each menstrual cycle or estrus cycle in humans or rodents, respectively. The investigators were the first to identify the stem cells that regenerate this tisse. As the epigenetic changes in estrogen response persist in the adult, they are likely to be encoded in the stem cells that regenerate the endometrium in each reproductive cycle. The investigators hypothesize that xenoestrogen exposure affects methylation of multiple EREs in uterine stem cells leading to altered estrogen sensitivity as an adult. To test this hypothesis the investigators will determine the extent of uterine ERE methylation in the entire genome after DES or BPA exposure as well as the effect of exposure to these agents on endometrial stem cell growth and estrogen response in vitro and in vivo.
The specific aims i nclude 1) the use of chromatin precipitation, massively parallel sequencing and bisulfite sequencing to determine if exposure leads to preferential methylation of multiple EREs and if this occurs in uterine stem cells;2) determine if exposure leads to altered endometrial stem cell estrogen response in vitro;and 3) use of an in vivo model to determine if endometrial stem cells from exposed animals are more prone to endometriosis or endometrial cancer. These studies will test the hypothesis that methylation of EREs in uterine stem cells and resultant altered estrogen responsiveness will lead to an increased incidence of estrogen mediated disorders, thus providing an epigenetic mechanism for reproductive tract disease associated with xenoestrogen exposure. This model explains how a weak estrogen results in an estrogenic response disproportionate to its intrinsic estrogenic activity and how this epigenetic signal persists despite loss of endometrium in each reproductive cycle due to altered methylation in endometrial stem cells.

Public Health Relevance

Stem cells regenerate the endometrial lining of the uterus which is shed with each menstrual period. These studies will determine if the effects of fetal environmental estrogen exposure are epigenetically encoded in the endometrial stem cells. Knowledge of the molecular and cellular mechanisms that underlie the effects of environmental estrogen exposure will allow the design of preventive and therapeutic strategies.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD076422-02
Application #
8528662
Study Section
Special Emphasis Panel (ZES1-TN-D (ST))
Program Officer
Ravindranath, Neelakanta
Project Start
2012-08-10
Project End
2017-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
2
Fiscal Year
2013
Total Cost
$395,021
Indirect Cost
$157,771
Name
Yale University
Department
Obstetrics & Gynecology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Naqvi, Hanyia; Mamillapalli, Ramanaiah; Krikun, Graciela et al. (2016) Endometriosis Located Proximal to or Remote From the Uterus Differentially Affects Uterine Gene Expression. Reprod Sci 23:186-91
Fischer, Catha; Mamillapalli, Ramanaiah; Goetz, Laura G et al. (2016) Bisphenol A (BPA) Exposure In Utero Leads to Immunoregulatory Cytokine Dysregulation in the Mouse Mammary Gland: A Potential Mechanism Programming Breast Cancer Risk. Horm Cancer 7:241-51
Jorgensen, Elisa M; Alderman 3rd, Myles H; Taylor, Hugh S (2016) Preferential epigenetic programming of estrogen response after in utero xenoestrogen (bisphenol-A) exposure. FASEB J 30:3194-201
Flannery, Clare A; Fleming, Andrew G; Choe, Gina H et al. (2016) Endometrial Cancer-Associated FGF18 Expression Is Reduced by Bazedoxifene in Human Endometrial Stromal Cells In Vitro and in Murine Endometrium. Endocrinology 157:3699-3708
Flannery, Clare A; Rowzee, Anne M; Choe, Gina H et al. (2016) Development of a Quantitative PCR Assay for Detection of Human Insulin-Like Growth Factor Receptor and Insulin Receptor Isoforms. Endocrinology 157:1702-8
Kulp, Jennifer L; Mamillapalli, Ramanaiah; Taylor, Hugh S (2016) Aberrant HOXA10 Methylation in Patients With Common Gynecologic Disorders: Implications for Reproductive Outcomes. Reprod Sci 23:455-63
Cosar, Emine; Mamillapalli, Ramanaiah; Ersoy, Gulcin Sahin et al. (2016) Serum microRNAs as diagnostic markers of endometriosis: a comprehensive array-based analysis. Fertil Steril 106:402-9
Flannery, Clare A; Saleh, Farrah L; Choe, Gina H et al. (2016) Differential Expression of IR-A, IR-B and IGF-1R in Endometrial Physiology and Distinct Signature in Adenocarcinoma. J Clin Endocrinol Metab 101:2883-91
Mutlu, Levent; Hufnagel, Demetra; Taylor, Hugh S (2015) The endometrium as a source of mesenchymal stem cells for regenerative medicine. Biol Reprod 92:138
Yang, Huan; Kang, Kai; Cheng, Chao et al. (2015) Integrative Analysis Reveals Regulatory Programs in Endometriosis. Reprod Sci 22:1060-72

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