Projects / Programmes
Role of progesterone metabolites in hormone dependent diseases
| Code |
Science |
Field |
Subfield |
| 3.07.00 |
Medical sciences |
Metabolic and hormonal disorders |
|
| Code |
Science |
Field |
| B000 |
Biomedical sciences |
|
| Code |
Science |
Field |
| 3.02 |
Medical and Health Sciences |
Clinical medicine |
progesterone, endometrial cancer, endometriosis, metabolites
Researchers (17)
Organisations (2)
Abstract
The hormone-dependent diseases including hormone-dependent cancers, which comprise more than 20% of all cancers, and benign hormone dependent diseases are extremely frequent. In the last decades studies on these diseases in women mainly focused on the role of estrogens, while the role of progesterone (P) did not get sufficient attention. Although the action of (P) metabolizing enzymes is generally viewed as means of reducing its local concentration, recent studies revealed important role of P metabolites in breast cancer. These metabolites, particularly 5a-pregnanes, are formed in increased levels in breast cancer tissue, where they activate membrane-bound receptors, and thus stimulate cell proliferation, migration, and decrease apoptosis. This role of P metabolites has been studied mainly in breast cancer, while there is no data for other types of hormone-dependent diseases, including endometrial cancer (EC) and endometriosis. EC is the most common gynecological malignancy and the fourth-most-common cancer in women in the developed world, with 30% of cases diagnosed in premenopausal women. EC is usually treated with hysterectomy while medical treatment of EC patients who want to preserve fertility is still a challenge. Endometriosis is a complex, polygenic disease that is defined as the presence of endometrial glands and stroma outside the uterine cavity. It is diagnosed in up to 15% of all premenopausal women and 35-50% of women with infertility. The classical medical treatments of endometriosis have adverse side effects and negative impacts on quality of life. There is thus an extreme need for new approaches for individualized treatment of EC and endometriosis. Based on our preliminary data we propose a hypothesis that 5a-pregnanes are formed in diseased endometrium of EC and endometriosis patients, where they can activate specific membrane-bound P receptors and intracellular signaling pathways, to stimulate proliferation and migration of cancer and endometriosis cells, and are thus implicated in pathogenesis. Aims of the project are as follows: 1) to clarify P metabolism in EC and endometriosis. We aim to examine the metabolism of P in EC and endometriosis tissue samples and model cell lines and to pinpoint individual enzymes with the use of specific inhibitors and siRNA approach. P metabolites formed will be identified by LC-MS/MS. 2) To explain mechanisms of actions of P metabolites in EC and endometriosis. We aim to evaluate the effects of P metabolites on cell proliferation, migration and invasion of the model cell lines in a real time scale. The activation of signalling pathways that may be responsible for the proliferative effects of 5a-pregnanes will also be studied. The mechanisms of actions of P metabolites will be delineated by determining the individual P receptors, and the components of the intracellular signalling cascades, further enabling identification of novel therapeutic strategies. This project will contribute to: 1) identification of novel drug targets (among P metabolizing enzymes and membrane P receptors and enzymes of intracellular signaling pathways); 2) development of diagnostic assays for individualized treatment (immunohistochemical staining of individual P metabolic enzymes or P receptors in biopsies of diseased endometrium); and may lay foundation for 3) development of noninvasive diagnostic assay (identification of P metabolites in plasma and urine of EC and endometriosis patients).
Significance for science
In the last decade studies on hormone-dependent diseases in women mainly focused on the role of estrogens, while the role of progesterone (P) has attracted less attention. P metabolites were considered as inactive compounds, until in the recent years 5alpha-pregnanes have been implicated in development of breast cancer, where they stimulate cell proliferation, migration, and decrease apoptosis. The role of P metabolites in endometrial cancer and endometriosis has not been studied, the metabolic enzymes and individual metabolites formed in diseased tissue have not been identified and also the mechanisms of their action have not been explained. Although several new types of membrane bound PR (mPR) have been found recently, their expression in diseased endometrium and their potential association with the action of P metabolites have not been examined. In the framework of this project we clarified P metabolism and the role of P metabolites in EC and endometriosis. We examined the mechanisms of actions of P metabolites and their effects on proliferation. The project contributed to recognition of P metabolizing enzymes, AKR1C2 and SRD5A1, as potential novel drug targets and mPRs as potential prognostic biomarkers. The LC-MS/MS method, which was established for separation and quantification of 16 P metabolites, enables analysis of the whole panel of P metabolites without previous derivatization, with good reproducibility and femtomolar sensitivity. This method can be used in clinical studies and may thus contribute to identification of P metabolites as diagnostic/ prognostic biomarkers of different pathological states. In general the results of our project expanded the knowledge in the field of molecular endocrinology and especially with regards to novel mechanisms of action of steroid hormones.
Significance for the country
The results of the research project were published and were presented at several international scientific conferences thus contributing to international affirmation and promotion of Slovenian science. This project enabled collaboration with esteemed institutions, Helmholtz Zentrum Munchen, Germany and University of Texas Austin, Port Aransas, USA and transfer of the cutting-edge knowledge on steroid analytics and methodological approaches to study membrane bound steroid receptors to Slovenia. The proposed project enhanced the quality of research work as well as the quality of teaching at the Faculty of Medicine of the University of Ljubljana. The knowledge stemming from this project has been disseminated to medical students through teaching Elective Classes Steroid Biochemistry and Hormone-dependent Diseases: Clinical and Molecular Aspects. In the framework of this research project two Ph.D. students were trained in the state-of-the-art methods for the identification and characterization of steroid metabolites, and in new mechanisms of steroid hormone actions. The good results of the proposed project will help the collaborating institutions, the Faculty of Medicine of the University of Ljubljana and the Department of Obstetrics and Gynaecology of the Medical Centre Ljubljana in terms of international recognition.
Most important scientific results
Annual report
2013,
2014,
2015,
final report
Most important socioeconomically and culturally relevant results
Annual report
2013,
2014,
2015,
final report
