Role of estrogen receptor beta in epithelia lacking estrogen receptor alpha expression

Estrogens influence the physiology of many organs by activation of the nuclear receptors ERalpha and ERbeta. The epithelium of ventral prostate, ovarian granulosa cells and lung, are ERalpha-negative but express high levels of ERbeta. This thesis focuses on the role played by ERbeta in ERalpha-negative epithelium.

The prostate is of medical interest because it is very frequently the site of malignancy in elderly men. Prostate cancer is the most lethal form of cancer in men after lung cancer. We studied the ventral prostate of mice to gain insight into the role of ERbeta in prostatic epithelium. We used histochemical, protein profiling and FACS analysis to study the ventral prostate of mice in which ERbeta gene has been inactivated (ERbeta−/− mice). We found epithelial hyperplasia, accumulation undifferentiated epithelial cells and dysregulation of several proteins associated to low differentiation level. Among these proteins, serine protease inhibitor Kazal type 3, called Spink3. The human homologous of Spink3, TATI, is positively related to low epithelial differentiation grade (Gleason Score) in human prostate cancer. Analyzing prostate cancer biopsies of patients at different Gleason Scores we observed a negative correlation between TATI and ERbeta expression and positive correlation between TATI and Gleason Score. This study showed a role for ERbeta in differentiation of the prostatic epithelium in mice and man.

The granulosa cells of the ovarian follicle are epithelial-derived cells which secrete estradiol. which is essential for maturation of the ovum, and preparation of the entire body for response to a fertilized ovum. In the ovarian follicles ERbeta is abundantly expressed in the nuclei of granulosa cells, while the theca cells are ERalpha-positive and ERbeta-negative. Theca cells are stromal-derived cells which form a highly vascularized layer surrounding the granulosa cells during follicular maturation. Female of ERbeta−/− mice vary rarely ovulate and are considered to be subfertile. In the ovary, there are very few corpora lutea and many atretic follicles. Upon wedge resection of the ovaries maturation of follicles and ovulation were restored, clearly indicating that the hypothalamo-pituitary axis in female ERbeta−/− mice is normal. Upon histochemical and immunohistochemical examination, we observed impaired vascularization in the theca cell layer of ovarian follicles in ERbeta−/− mice. We hypothesized that ERbeta expression in granulosa cells is important for the cross-talk between granulosa and theca cell compartments, in order to allow vascularization of theca layers during follicular maturation. We further conclude that the trauma of ovarian wedge resection can trigger activation of angiogenic signaling and overcome the lack of ERbeta functions.

The major function of the lung is to allow gas exchanges between blood stream and the air. In order to perform such a function, the main characteristics of lung are to provide large surface availability and elasticity. These two features depend respectively by the structure of alveoli and the active turnover of the extracellular matrix. The lung alveolar and bronchiolar epithelium are ERbeta-positive. ERbeta−/− mice are hypertensive and compared to normal mice, their lungs have large alveoli and reduced elastic recoil. Our investigation of the lungs of ERbeta-/- mice provide evidence that ERbeta is involved in the regulation of key proteins important for the control of extracellular matrix (ECM) in the lung parenchyma and that ERbeta−/− mice are hypoxic and reluctant to run on the treadmill. We speculate that systemic hypoxia can be one of the causes of systemic hypertension characteristic of ERbeta−/− mice.

List of scientific papers

I. Imamov O, Morani A, Shim GJ, Omoto Y, Thulin-Andersson C, Warner M, Gustafsson JA (2004). Estrogen receptor beta regulates epithelial cellular differentiation in the mouse ventral prostate. Proc Natl Acad Sci USA. 101(25): 9375-80. Epub 2004 Jun 8
https://pubmed.ncbi.nlm.nih.gov/15187231

II. Schwend T, Imamov O, Morani A, Ruppert T, Waage L, Zickert P, Bergerheim U, Warner M, Gustafsson JÅ (2008). Identification of spinK3 as indicator of ER beta negative, poorly differentiated prostatic epithelium by isotope coded protein labeling and LC-ESI-MS/MS. [Submitted]

III. Inzunza J, Morani A, Cheng G, Warner M, Hreinsson J, Gustafsson JA, Hovatta O (2007). Ovarian wedge resection restores fertility in estrogen receptor beta knockout (ERbeta-/-) mice. Proc Natl Acad Sci USA. 104(2): 600-5. Epub 2006 Dec 29
https://pubmed.ncbi.nlm.nih.gov/17197418

IV. Morani A, Barros RP, Imamov O, Hultenby K, Arner A, Warner M, Gustafsson JA (2006). Lung dysfunction causes systemic hypoxia in estrogen receptor beta knockout (ERbeta-/-) mice. Proc Natl Acad Sci USA. 103(18): 7165-9. Epub 2006 Apr 24
https://pubmed.ncbi.nlm.nih.gov/16636272