IMPACT OF DIETARY ESTROGENS FROM SOY AND OTHER CROPS ON REPRODUCTIVE AND METABOLIC HEALTH OF WOMEN

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: TERMINATED
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 1004548
• Project No.: ILLU-698-909
• Project Start Date: Dec 9, 2014
• Project End Date: Sep 30, 2019

Project Director
Madak-Erdogan, ZE, .

Recipient Organization
UNIVERSITY OF ILLINOIS
2001 S. Lincoln Ave.
URBANA,IL 61801

Performing Department
Food Science & Human Nutrition

Non Technical Summary
Health maintenance, life quality, and resilience of postmenopausal women remain major issues because, due to advancements in healthcare, by 2030 more than 25% of the US population will be over 60, and most will be women. Epidemiological studies suggest that the combination of loss of estrogens after menopause, western diet, and sedentary life style increases the incidence of obesity and associated metabolic syndrome and cardiovascular disease, and may alter breast cancer progression. Incidence of obesity, diabetes, cardiovascular disease and cancer is even higher in rural areas and all of the aforementioned diseases were identified as rural health priorities by both national and state experts. Hormone-replacement therapies (HRT) provide benefits to menopausal women but increase the risk of breast and endometrial cancers. As alternatives to HRT, botanical estrogen dietary supplements (BEDS) from soy and other plants are used by many women with the assumption that because these products are "natural", they are inherently safe and will promote health and resiliency. This impacts the level of demand on natural products, especially of soy which is a main crop that is cultivated in USA. Therefore, it is of great importance (a) to evaluate the tissue-selective activities of BEDS taken by postmenopausal women and to determine how these might affect their health, and (b) to determine whether BEDS and dietary factors impact quality of life and resilience to obesity and CVD.

Animal Health Component

0%

Research Effort Categories

Basic

50%

Applied

50%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
702	1820	1010	50%
702	2410	1040	50%

Knowledge Area
702 - Requirements and Function of Nutrients and Other Food Components;

Subject Of Investigation
1820 - Soybean; 2410 - Cross-commodity research--multiple crops;

Field Of Science
1040 - Molecular biology; 1010 - Nutrition and metabolism;

Keywords

reproductive health

metabolic health

dietary estrogens

Goals / Objectives
Adding IACUC Approval to active project on 12/09/2014.The objective of this project is to characterize different BEs and identify BEs with beneficial effects in non-reproductive metabolic tissues vs. risks to reproductive tissues, including stimulation of endometrium and increased metastasis of BC. This project will further evaluate the mechanisms for interactions of BEs with the development of metabolic syndrome and obesity associated with menopause and high fat diets. Ourlong-term goal is to providean in vivo assessment of the selective actions of BEs in preclinical mouse models relevant to the metabolic health of menopausal women and BC survivors. This project will study the effects of BEs from wild yam, licorice root, and Epimedium, as multi-component mixtures, fractions and isolated components, on important reproductive and metabolic target tissues in female mice, including uterus, mammary gland, liver, various fat depots, and skeletal muscle.Overall aim: Determine the effects of BEs on reproductive and non-reproductive tissues, and on metabolism in mouse models of menopause-associated metabolic syndrome.Specific Aim 1: Establish the effects of BEs on metabolism after ovariectomy and high fat diet.a. Total body weight, uterus, and adipose tissue weightsb. MRI analysis of fat contentc. Food consumptiond. Histological assessment of liver, adipose tissue, skeletal musclee. Histological assessment of uterus and mammary glandSpecific Aim 2: Assess the effects of BEs and high fat diets on endogenous metabolite and biomarker profiles in blood.a. Blood metabolic profilingb. Triglyceride levels and lipid profile in liver and bloodc. Cancer and cardiovascular disease biomarker levels in the bloodSpecific Aim 3: Determine molecular effects of BEs and high fat diets in liver, various fat depots, skeletal muscle, uterus and mammary gland.a. Gene expression analysis by RNA-Seq in liver, adipose tissues, uterus, mammary gland, skeletal muscle, primary hepatocytes and breast cancer cellsb. ERα, LXRα/β, ERK2, Raptor, RNA Pol II CHIP-Seq (liver, uterus, adipose tissues, primary hepatocytes and breast cancer cells)c. mTOR, AMPK, p42/44, p38 and JNK MAPK pathways activation in all tissuesOur findings from metabolic studies done in wild-type and knockout animals (Specific Aim 1), the blood metabolite and biomarker profiling (Specific Aim 2) combined with the gene expression profiles and pathway analyses in different target tissues (Specific Aim 3) should reveal aspects of the underlying molecular mechanisms that might account for any tissue-selective effects we observe with the BEs.

Project Methods
Animals and diets - To test the impact of the BEs on resiliency to obesity and metabolic syndrome, we will use a high fat diet and compare it to a control diet. The normal chow contains isoflavones, which can have estrogenic activities that could abrogate the metabolic effects of ovariectomy. All studies in this aim will use a semi-purified (AIN76A) diet having casein as the protein source. The high fat diet for Aim 1 of this study (Harlan TD.88137) is termed "western diet" and is based on an AIN76A diet (45% Kcal from fat). It is high in butterfat with sugar and some cholesterol (0.2%), and it was developed for studies of metabolic syndrome.Specific Aim 1: Establish the effects of BES on metabolism after ovariectomy and high fat diet.Metabolic studies- In order to assess the impact of BES on gene expression profiles in ovariectomized animals, 6 weeks old C57BL6 mice will be divided into two groups. One group will continue in normal diet (AIN76A) whereas the second group will be placed in high fat diet for 2 weeks, prior to ovariectomy. After 3 weeks, pellets containing E2, BEs with doses identified from administration studies will be implanted subcutaneously. Weight of the animals, food intake and level of compounds in the blood will be assessed every week. MRI analysis will be performed on 5 weeks post treatment groups to determine fat and lean mass of the animals. At the end of the 5th week blood, uterus, mammary gland, liver, inguinal fat, perigonadal fat, perirenal fat, mesenteric fat, brown adipose, and skeletal muscle will be harvested. Duration of study, 5 weeks, is based on our previous studies. 5 weeks is early enough to determine the molecular changes in the organs studied that are associated with high fat diet and ovariectomy. We will characterize the histological and molecular properties of the tissues harvested, and correlate these with metabolic phenotype.Specific Aim 2: Assess the effects of BES and high fat diets on endogenous metabolite and biomarker profiles in blood.Our previous metabolomics analysis with botanical estrogens and low affinity ER ligands showed that these compounds induced changes in amino acid metabolites, especially glutamine metabolism, and TCA cycle metabolites in the blood of animals. Based on these findings, we examined glutamine metabolism genes in the main sites of glutamine metabolism, namely, liver, skeletal muscle, and adipose tissues. Our results indicated that while E2 and low affinity ER ligands up-regulated expression of glutamine synthesis enzymes, they down-regulated expression of glutamate synthesis enzymes in skeletal muscle but not in the liver. These results show that low affinity estrogens modulated the glutamine metabolism pathway and levels of a number of metabolites in blood in a tissue-selective manner. These changes are consistent with an improved metabolic profile, and they suggest that other low-affinity ligands, such as BEs, might have similar beneficial effects. Thus, we will use a similar approach to define the effects of all of the BES on metabolic parameters. To compare metabolic changes associated with BEs supplementation after ovariectomy mice, we will harvest blood from animals in Specific Aim 1 at the end of 3 weeks and perform a complete metabolic profiling at the University of Illinois Metabolomics Core. The levels of compounds obtained after mass spec analysis will be clustered using Cluster 3 software and visualized using Java Tree View; identified groups of compounds will further be analyzed using Metacore Software. Levels of lipids (LDL, HDL, total cholesterol and Triglycerides) will also be monitored. Additionally, we will monitor triglyceride level in the liver after ligand treatments. Finally, we will use Proseek Multiplex CVD, the largest and most comprehensive pre-validated CVD panel from OLINK Bioscience to assess levels of some CVD and inflammation markers and evaluate the impact of BES on expression of these markers.Specific Aim 3: Determine molecular effects of BES and high fat diets in liver, various fat depots, and skeletal muscle.In our previous studies we found that E2 and low affinity ER ligands resulted in similar profiles in metabolic tissues, whereas only E2 induced gene expression changes in uterus. To further characterize BES induced molecular changes in the various tissues harvested from animals in metabolic experiments, we will first perform gene expression analysis in liver, several fat depots, skeletal muscle, pancreas and uterus. For this study we will use tissues harvested from metabolic study animals from Specific Aim 1.For ChIP-Seq analysis 6 weeks old C57BL6 mice will be divided into two groups. One group will continue in normal diet (AIN76A) whereas the second group will be placed in high fat diet for 3 weeks prior to ovariectomy. After 3 weeks, female mice will be injected intraperitoneally proper doses of E2 or BEs for 2 hours. Animals will be sacrificed and the following tissues will be harvested: liver, uterus, skeletal muscle, inguinal fat, perigonadal fat, mesenteric fat, perirenal fat, and mammary gland. Chromatin will be prepared from the resulting tissues.Transcriptome and cistrome analysis - In this study we are proposing to perform sequencing-based transcriptome and cistrome analysis in liver, fat depots (inaugural, perigonadal, mesenteric, perirenal and brown adipose), skeletal muscle, uterus and mammary gland tissues harvested from animals in the metabolism study. We have extensive previous experience in conducting RNA-Seq and ChIP-Seq experiments and analyzing large datasets. We also have preliminary data from studies using low affinity estrogens and some botanical estrogens. Samples will be submitted to the UIUC Sequencing Core facility for library preparation and sequencing. We have extensive experience using several well-established, integrated pipelines for the analysis of sequencing data.Signaling pathway analysis - We performed western blot analysis to profile which signaling pathways were utilized by E2 and low affinity ligands in different tissues. Our results showed that E2 was effective in activating MAPK and mTOR signaling in reproductive and non-reproductive tissues. However, the low affinity ligands activated MAPK and mTOR only in the non-reproductive tissues. Therefore: 1) E2 and low affinity ligands have similar effects on non-reproductive tissues where estrogen action is needed to regulate important beneficial metabolic effects; 2) E2 and low affinity ligands have opposite effects in reproductive tissues where proliferative effects are detrimental, with only E2 being a stimulator; and 3) In metabolic tissues like liver and skeletal muscle, both E2 and low affinity estrogens were highly effective in inducing mTOR signaling, as observed by S6 phosphorylation.These experiments have nicely contrasted the tissue-selective cell regulatory effects of E2 and low affinity estrogens, and thus they form a conceptual template that we will use to evaluate the mechanistic bases for selective effects of the different BEs that we will be studying. For signaling pathway analysis, 6 weeks old C57BL6 mice will be divided in to two groups. One group will receive normal diet (AIN76A) whereas the second group will be placed in high fat diet for 2 weeks prior to ovariectomy. After 3 weeks of wash out period, female mice will be injected intraperonitenally with appropriate doses of E2 or BEs for 2 hours. Animals will be sacrificed and the following tissues will be harvested: liver, uterus, skeletal muscle, inguinal fat, perigonadal fat, mesenteric fat, perirenal fat, bat, mammary gland, kidney, and spleen. Protein extract will be prepared from all tissues. We will be particularly attentive to effects of BEs in correlating their biological effects in vivo with these molecular changes in cell regulatory signaling pathways.

Progress 12/09/14 to 09/30/19

Outputs
Target Audience:The main stakeholders are scientists in the fields of women's health, agriculture, and environmental toxicology and students. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Eylem Kulkoyluoglu finished her Ph.D studies and graduated in May 2019. She is now a postdoctoral fellow at Indiana University. Kevin Dong, an undergraduate student who was involved in this project, graduated last year. He was accepted at MD. Anderson Cancer Biology Ph.D. program. How have the results been disseminated to communities of interest?Several manuscripts were published. In addition, PI and graduate students gave numerous talks at national meetings (Endocrine Society Annual Meeting, American Association for Cancer Research Annual Meeting), reporting findings from this project. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Specific Aim Two: We found that obesity increased free fatty acid levels in the blood. These fatty acids worked with estrogen receptors to increase risk of breast cancer. Using pathway preferential estrogens we were able to reduce level of free fatty acids in the circulation. In addition, these estrogens rewired breast cancer cell metabolism to negate the effects of free fatty acids. Fatty acids incudes previously unknown interactions between estrogen receptor and PPARs. These interactions resulted in metabolic rewiring of breast cancer cells. Overall, our findings from this study were able to explain why obesity is a risk factor for estrogen receptor positive breast cancer. Other aims were completed in previous years.

Publications

• Type: Journal Articles Status: Published Year Published: 2019 Citation: Madak-Erdogan, Z., Band, S., Zhao, Y.C., Smith, B.P., Kulkoyluoglu-Cotul, E., Rossi, G., Kim, S.H., Wrobel, K., Smith, R., Johnson, M.L., Patel, M., Marino, N., Storniolo, A.M.V., Katzenellenbogen, J.A. and Flaws, J.A. 2019. Free fatty acids rewire cancer metabolism in obesity-associated breast cancer through Estrogen Receptor and mTOR signaling, Cancer Research, 2019 Mar 12. doi: 10.1158/0008-5472.CAN-18-2849.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Kulkoyluoglu-Cotul, E., Arca, A. and Madak-Erdogan, Z. 2019. The crosstalk between estrogen signaling and breast cancer metabolism. Trends in Endocrinology and Metabolism. Jan. 30(1):25-38 doi: 10.1016/j.tem.2018.10.006 PMID: 30471920.
• Type: Journal Articles Status: Published Year Published: 2019 Citation: Kulkoyluoglu-Cotul, E., Smith, B.P., Wrobel, K., Zhao, Y.C., Chen, K.L., Hieronymi, K., Imir, O.B., Duong, K., OCallaghan, C., Mehta, A., Sahoo, S., Haley, B., Chang, H., Landesman, Y., Madak-Erdogan, Z. 2019. Combined targeting of estrogen receptor alpha and XPO1 prevent Akt activation, remodel metabolic pathways and induce autophagy to overcome tamoxifen resistance. Cancers (Basel). April 4;11(4). pii: E479. doi: 10.3390/cancers11040479. PMID: 30987380.
• Type: Journal Articles Status: Under Review Year Published: 2019 Citation: Oktay, K., Santaliz-Casiano, A., Johnson, M.L., Patel, M., Marino, N., Storniolo, A.M.V., Torun, H., Acar, B. and Madak-Erdogan Z. 2019. A machine learning based evaluation of blood serum biomarkers for breast cancer risk assessment. In Revision BMC Cancer (Impact factor 3.28).

Progress 10/01/17 to 09/30/18

Outputs
Target Audience:The target audience includedscientists in the field of metabolic regulation and breast cancer. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project resulted in publications for several graduate students. They presented findings from these studies in several conferences including the AACR annual meeting and the Endocrine Society Annual Meeting. As part of these meetings, trainees attended early career workshops. How have the results been disseminated to communities of interest?Several manuscripts are under review and in progress. Also, the PI gave several talks atvarious conferences and universities. Trainees presented their findings in conferences. What do you plan to do during the next reporting period to accomplish the goals?We will continue our studies with developing an obesity and breast cancer animal model where we will test some of our findings from Aims 2 and 3.

Impacts
What was accomplished under these goals? Specific Aim 2: Assess the effects of BEs and high fat diets on endogenous metabolite and biomarker profiles in blood: We found that obesity increased free fatty acid levels in the blood. These fatty acids worked with estrogen receptors to increase risk of breast cancer. Using pathway preferential estrogens we were able to reduce level of free fatty acids in the circulation. In addition, these estrogens rewired breast cancer cell metabolism to negate the effects of free fatty acids. We have now completed this aim and submitted two publications containing this data. Specific Aim 3: Determine molecular effects of BEs and high fat diets in liver, various fat depots, skeletal muscle, uterus and mammary gland. The first step of this aim is completed. RNA-Seq analysis of livers from mice treated with vehicle or pathway preferential estrogens showed that estrogen supplementation increased mitochondrial biogenesis and decreased collagen deposition. Net effect was an overall decrease in liver lipid deposition, decrease fibrosis of liver and normalization of blood metabolites associated with obesity. We are in the process of preparing a manuscript containnig results from Specific Aim 3a.

Publications

• Type: Journal Articles Status: Published Year Published: 2018 Citation: Chen, K.L., Liu, X., Zhao, Y., Hieronymi, K., Rossi, G., Auvil, L.S., Welge, M., Bushell, C., Smith, R.L., Carlson, K.E., Kim, S.H., Katzenellenbogen, J.A., Miller, M.J. and Madak-Erdogan, Z. 2018. Long-Term Administration of Conjugated Estrogen and Bazedoxifene Decreased Murine Fecal Beta-Glucuronidase Activity Without Impacting Overall Microbiome Community. Scientific Reports 2018 May 25;8(1):8166. doi: 10.1038/s41598-018-26506-1, PMID: 29802368.
• Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Madak-Erdogan, Z., Band, S., Zhao, Y.C., Smith, B.P., Kulkoyluoglu-Cotul, E., Rossi, G., Kim, S.H., Wrobel, K., Smith, R., Johnson, M.L., Patel, M., Marino, N., Storniolo, A.M.V., Katzenellenbogen, J.A. and Flaws, J.A. 2018. Free Fatty Acids Rewire Cancer Metabolism In Obesity-Associated Breast Cancer Through Estrogen Receptor and mTOR Signaling. Corresponding author, in revision Cancer Research.
• Type: Journal Articles Status: Under Review Year Published: 2018 Citation: Oktay, K., Santaliz-Casiano, A., Zhao, Y.C., Wrobel, K., Johnson, M.L., Patel, M., Marino, N., Storniolo, A.M.V., Flaws, J., Torun, H., Acar, B. and Madak-Erdogan, Z. 2018. A Machine Learning Based Evaluation of Blood Serum Biomarkers for Breast Cancer Risk Assessment. Under revision BMC Cancer.

Progress 10/01/16 to 09/30/17

Outputs
Target Audience:Members of the target audience included cancer survivors and our peers in the field of nutrition. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?Systems Biology of Gene and Metabolic Regulation by Estrogen Receptors and Kinases in Breast Cancer and Metabolic Disease, Purdue University, September, Lafayette, Indiana, invited. Systems Biology of Gene and Metabolic Regulation by Estrogen Receptors and Kinases in Breast Cancer and Metabolic Disease, Carle Cancer Symposium, September 2016, Urbana, IL, invited. Systems Biology of Gene and Metabolic Regulation by Estrogen Receptors and Kinases in Breast Cancer and Metabolic Disease, University of Texas Southwestern (UTSW) Medical Center, Department of Reproductive Biology Seminar, October 2016, Dallas, TX, invited. Extra-Nuclear ERα Signaling in Rewiring Cancer Cell Metabolism During Obesity-Associated Postmenopausal Breast Cancer, Jensen Symposium on Breast Cancer, November 2016, Cincinnati, OH, invited. Biomarker Identification and Integrative -Omics Approaches to Understanding Breast Cancer Disparities, University of Illinois, Chicago, Breast Cancer Research Program, Research Symposium Speaker, February 2017, Chicago, IL, invited. Extra-Nuclear ERα-mTOR Signaling Rewires Cancer Cell Metabolism During Obesity-Associated Breast Cancer, AACR Annual Meeting, Washington DC, April 2017. Systems Biology of Estrogen Receptor-Kinase Signaling Crosstalk in Cancer and Metabolism, FASEB Conference on Rapid Signaling andGenomic Hormone Action in Health andDisease, June 2017, Snowmass Village, CO , invited Role of Nuclear Transport Pathways in Development of Tamoxifen Resistance in Breast Cancer, Gordon Conference on Hormone Dependent Cancers, August 2017, Maine, invited. Development of Multi-Scale Analysis Methods to Understand the Role of Estrogen Receptors and Kinases in Breast Cancer and Metabolic Syndrome, NIH Future Research Leaders Conference, September 2017, Bethesda, MD. Systems Biology of Extranuclear-ERα Initiated Kinase Signaling in Breast Cancer and Metabolism, FASEB Conferences on Rapid Signaling and Genomic Hormone Action in Health & Disease, July 2019, Organizer and speaker . What do you plan to do during the next reporting period to accomplish the goals?During the next reporting period we will continue our molecualr analysis.

Impacts
What was accomplished under these goals? We were able to complete the following Objectives: Specific Aim 2: Assess the effects of synthetic estrogens and high fat diets on endogenous metabolite and biomarker profiles in blood. a. Blood metabolic profiling andb. Cancer and cardiovascular disease biomarker levels in the blood. Specific Aim 3: Determine molecular effects of synthetic estrogens and high fat diets in liver, various fat depots, skeletal muscle, uterus and mammary gland.a.Gene expression analysis by RNA-Seq in liver, adipose tissues, uterus, mammary gland, skeletal muscle, primary hepatocytes and breast cancer cells. Specifically,We were able to complete molecular characterization of tissue from wildtype and ob/ob mice that were treated with various estrogens. These analyses showed that estrogens mainly impacted lipid deposition pathways in liver and reduced expression of oxidative stress-associated genes. These protective effects in the liver were through ERalpha as these responses were not repeated in cell line models which do not express ERalpha.Metabolomics analysis of the plasma from the same animals showed that the levels of free fatty acids and certain amino acids were increased in animals which were obese. Treatment with estrogens normalized these metabolites to the levels similar to that of lean animals.We monitored levels of cardiovascular disease, inflammation and cancer biomarkers using serum samples from postmenopausal women. These analyses identified differences in inflammation markers.

Publications

• Type: Journal Articles Status: Under Review Year Published: 2017 Citation: K.L. Chen, Y.C. Zhao, B. Smith and Z. Madak Erdogan. 2017. Bazedoxifene and conjugated estrogen combination maintains metabolic homeostasis and benefits liver health. PLOS ONE (In Revision).

Progress 10/01/15 to 09/30/16

Outputs
Target Audience:We have given several talks atnational and internationational conferences and universities to disseminate our research. We also started a Cancer Research Advocacy Group to train cancer patient advocates. Changes/Problems:Since the technology used for blood metabolite profiling is for samples from humans, we had to use samples from postmenopausal women who were obese and non-obese. Then we extrapolated these results to our mice studies. What opportunities for training and professional development has the project provided?Twograduate students presented their research at prestigious conferences including the Keystone Conference and the American Association for Cancer Research Annual Meeting. They also attended workshops related to nutrition and nutrigenomics. We also trained four undergraduate students, one postdoctoral fellow and one research technician. How have the results been disseminated to communities of interest?The published journal articles were announced on ourlab website and on Twitter and Linkedin. Several talks were given at conferences and universities. What do you plan to do during the next reporting period to accomplish the goals?We will continue our efforts to finish the sub aims of Specific Aims 2 and 3.

Impacts
What was accomplished under these goals? We have completed the following part for the Specific Aims below: Specific Aim 2: Assess the effects of low affinity estrogens and high fat diets on endogenous metabolite and biomarker profiles in blood. a. Blood metabolic profiling b. Cancer and cardiovascular disease biomarker levels in the blood (this was done for post menopausal women) Specific Aim 3: Determine molecular effects of low affinity estrogens and high fat diets in liver, various fat depots, skeletal muscle, uterus and mammary gland. a. Gene expression analysis by RNA-Seq in liver and in breast cancer cells

Publications

• Type: Journal Articles Status: Published Year Published: 2016 Citation: Chambliss, K.L., Barrera, J., Umetani, M., Umetani, J., Kim, S.H., Madak-Erdogan, Z., Katzenellenbogen, B.S., Katzenellenbogen, J.A., Mineo, C. and Shaul, P.W. 2016. Non-Nuclear Estrogen Receptor Activation Improves Hepatic Steatosis in Female Mice. Endocrinology 2016 Aug 23, PMID: 27552247.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Zhao, Y.C. and Madak Erdogan, Z. 2016. Systems Biology of Metabolic Regulation by Estrogen Receptor Signaling in Breast Cancer. Journal of Vis. Exp. 2016 Mar 17, PubMed PMID: 27023311.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Wrobel, K.U., Zhao, Y.C., Kulkoyluoglu, E., Chen, K.L., Hieronymi K., Holloway, J., Li, S., Ray, T., Ray, P.S., Lipka, A.E., Smith, R.L. and Madak-Erdogan, Z. 2016. XPO1 Regulates Tamoxifen Responsiveness in Estrogen Receptor Positive Breast Tumors by Decreasing Nuclear Localization of ERK5. Mol. Endocrinol. 2016 Aug. PMID:27533791. Featured in the journal website, selected for cover story, covered by UIUC website, EurekaAlert, News-Medical, Scifeeds and the Big Ten Network.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Madak-Erdogan, Z., Kim, S.H., Gong, P., Zhao, Y.C., Zhang, H., Chambliss, K.L., Carlson, K.E., Mayne, C.G., Shaul, P.W., Korach, K.S., Katzenellenbogen, J.A. and Katzenellenbogen, B.S. 2016. Design of Pathway Preferential Estrogens That Provide Beneficial Metabolic and Vascular Effects Without Stimulating Reproductive Tissues. Science Signaling 2016 May 24;9(429):ra53. PubMed PMID: 27221711. Featured in Editors Highlights, selected for cover story and a podcast by the journal.

Progress 12/09/14 to 09/30/15

Outputs
Target Audience:Our target audience is primarilyother scientists in the field that accessed our publication and attended our seminars. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?We have trained two undergraduate students, one graduate student, and one research assistant. How have the results been disseminated to communities of interest?The paper was announced in the laboratory website and on twitter. What do you plan to do during the next reporting period to accomplish the goals?We will continue our studies with the second and third aims.

Impacts
What was accomplished under these goals? We have accomplished allSpecific Aim 1 goals. Specifically we have established the effects of BEs on metabolism after ovariectomy and high fat diet. We monitored the following parameters: a.Total body weight, uterus, and adipose tissue weights; b. MRI analysis of fat content; c.Food consumption; d.Histological assessment of liver, adipose tissue, and skeletal muscle; and e.Histological assessment of uterus and mammary gland. We showed that licorice root supplementation prevented low estrogen level associated weight gain and increased lipid deposition. Findings from these studies were published in a manuscript entitled "Dietary licorice root supplementation reduces diet-induced weight gain, lipid deposition, and hepatic steatosis in ovariectomized mice without stimulating reproductive tissues and mammary gland".

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: Madak-Erdogan, Z., Gong, P., Zhao, Y.C., Xu, L., Wrobel, K.U., Hartman, J., Wang, M., Cam, A., Doerge, D., Khan, I.A., Katzenellenbogen, J.A., Katzenellenbogen, B.S. and Helferich, W.G. 2015. Dietary licorice root supplementation reduces diet-induced weight gain, lipid deposition, and hepatic steatosis in ovariectomized mice without stimulating reproductive tissues and mammary gland. Mol. Nutr. Food Res. 2015, 00, 112.