Oestrogen receptors and reproductive competence

Lead Research Organisation: MRC Human Reproductive Sciences Unit

Abstract

Oestrogens are steroids that are made in the testes and ovaries of men and women. They not only regulate our fertility but also have a major impact on our cardiovascular system, our brain, our bones and our immune system. Our quest to understand how sex steroids alter cell function has focused upon investigating the expression and functional activity of binding proteins (receptors) that are found in reproductive organs and cancers. In collaboration with clinical colleagues we have used antibodies to map the pattern of expression of oestrogen receptors (ERa and ER_) in normal human tissues such as the testis, ovary and the endometrial lining of the womb and also in cancers of the breast, colon and endometrium. Our current studies are using a variety of cell culture based assays and fluorescent protein imaging, to work out what impact oestrogens have on immune cells, cancers and fertility. We will be considering both the behaviour of receptors following binding to steroids and also what happens when they are activated by signals induced by growth factors. We hope that the results obtained may assist us in developing new approaches to the treatment of infertility and pathologies of the reproductive system.

Technical Summary

Oestrogens are synthesised within the gonads of adult men and pre-menopausal women. These steroids are essential regulators of fertility and also have an impact on the cardiovascular system, the immune system, the skeleton, the breast and the brain. Oestrogen action is mediated via specific nuclear receptors (ER). Two receptors known as oestrogen receptor alpha (ERalpha) and oestrogen receptor beta (ERbeta) are encoded by the ESR1 and ESR2 genes respectively. They belong to a superfamily of genes that includes three closely related orphan receptors know as oestrogen-related receptors alpha (ERRalpha), beta (ERRbeta) and gamma (ERRggamma). We have recently documented expression of ERRalpha and beta in human endometrium and obtained data suggesting that the long form of ERRbeta can modify ERalpha-dependent gene activation.|ERalpha and ERbeta act as ligand-activated transcription factors and can form homo- or heterodimers. ERs tagged with fluorescent proteins cloned into viral vectors are being used to investigate homo- and hetero-dimer formation in vitro using confocal imaging techniques (FRET, FRAP) and reporter assays with receptor subtype selective ligands (SERMS). We are also using cell-based approaches to investigate the impact of steroid ligand-independent pathways on receptor activity and gene expression. |In collaboration with Professory Hilary Critchely (University of Edinburgh) we have been studying the impact of oestrogens and androgens on cells within the human endometrium, a tissue that undergoes dynamic, cyclical remodelling in response to steroid hormones. The endometrium hosts a diverse group of immune cells including a phenotypically distinct population of natural killler [uNK] cells that that congregate at the maternal-fetal interface and play an essential role in establishment of pregnancy. We have discovered that these CD56+ cells express ERbeta but not ERalpha and Ilumina array analysis has revealed that transient exposure to oestradiol has a direct impact on their pattern of gene expression. |Our studies have shed new light on the role played by ERs and ERRs in the regulation of human fertility and in pathologies of the reproductive tract allowing the development of new treatment strategies.

Publications

10 25 50

publication icon
Childs AJ (2008) Modelling germ cell development in vitro. in Molecular human reproduction

publication icon
Critchley HO (2009) Hormone receptor dynamics in a receptive human endometrium. in Reproductive sciences (Thousand Oaks, Calif.)

 
Description Diabetes UK: RD Lawrence Personal Fellowship
Amount £157,316 (GBP)
Organisation Diabetes UK 
Sector Charity/Non Profit
Country United Kingdom
Start  
 
Description Society for Cell Biology Summer Studentships/British Society for Cell Biology
Amount £1,800 (GBP)
Organisation British Society for Cell Biology 
Sector Learned Society
Country United Kingdom
Start 06/2010 
End 09/2010
 
Description Wellcome Trust Project Grant
Amount £334,379 (GBP)
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 04/2008 
End 04/2011
 
Title Antibody 
Description We were the first group in the world to develop antibodies directed against oestrogen receptor beta. After our initial studies using our polyclonal antibody we collaborated with colleagues at Oxford Brookes University to develop monoclonal antibodies directed against wild type (full length) ERbeta as well as 2 truncated variants - the monoclonals have been commercialised 
Type Of Material Antibody 
Year Produced 2006 
Provided To Others? Yes  
Impact These antibodies have been widely used - the polyclonal antibodies opened up new avenues for research in male reproductive biology. The monoclonals have been used by resesarchers working on cancers of the breast, colon, prostate and endometrium. They have advanced our understanding of the impact of ER-dependent signalling on disease progression and suggested new avenues for therapy 
 
Description Bayer-Schering pharmaceuticals 
Organisation Bayer
Country Germany 
Sector Private 
PI Contribution We have used our expertise and available human cell lines to test the impact of an ERbeta selective agonist on receptor mobility and transcriptional activity
Collaborator Contribution Provided novel ERbeta agonists free of charge - this reagents are not available on the open market and are in development as therapeutics
Impact This reagent has given us novel insight into receptor dynamics that are relevant to reproductive disorders. Informal discussions and an invitation to speak at a workshop in Berlin.
Start Year 2009
 
Description Studies on androgen receptor promoter interactions 
Organisation Catholic University of Louvain
Country Belgium 
Sector Academic/University 
PI Contribution We have provided expertise in reproductive biology enabling the interpretation of mouse models
Collaborator Contribution Provision of reagents that have enhanced our research activity, insight into the regulation of gene expression by androgens that have informed our research plans
Impact Publications [17360365, 19587329]
 
Description Studies on breast cancer 
Organisation University of Leeds
Department Leeds Institute of Molecular Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution My team provided antibodies, insight into the activities of oestrogen receptors and it was my idea to examine the expression of the ERbeta5 variant in breast cancer tissues
Collaborator Contribution The Leeds group have access to an extensive archive of breast cancer tissues from patients with known outcomes. Analysis of oestrogen receptor expression in these tissues has advanced our understanding of the role(s) or ER-dependent signalling in disease and resulted in new directions in our research
Impact Publications [18698041, 19001343]
 
Description Studies on human endometrium 
Organisation University of Edinburgh
Department Centre for Reproductive Biology
Country United Kingdom 
Sector Academic/University 
PI Contribution My team provides intelectual input, novel reagents and molecular knowhow.
Collaborator Contribution Extensive access to human clinical material, clinical know how and insight into reproductive disorders, joint student supervision, joint laboratory activity
Impact Joint grant application to Wellcome Trust - successful. Joint publications [18032694, 18775884, 19196802, 19208787, 19864448, 19955102, 20885978, 20668045 ] IP - patent submitted but not supported beyond first year
 
Description Studies on uterine immune cells 
Organisation University of Edinburgh
Department MRC Centre for Inflammation Research
Country United Kingdom 
Sector Public 
PI Contribution Primary tissue, array analyses, functional assays, informatic analysis of data
Collaborator Contribution Access to equipment for cell isolation, training in the use of equipment and advice on immune cell phenotypes plus ideas and advice on phenotyping of cells
Impact Novel data that is being prepared for publication
Start Year 2008
 
Title ERbeta5 antibody 
Description A monoclonal antibody directed against a novel receptor splice variant over-expressed in cancers 
Type Diagnostic Tool - Non-Imaging
Current Stage Of Development Small-scale adoption
Year Development Stage Completed 2009
Development Status Under active development/distribution
Impact This antibody has been used to document expression in cancers of breast, colon and endometrium. The expression of this protein can give insight into cancer prognosis. The antibody has been commercialised 
 
Description Drama writers visit 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Organised and hosted a visit by Drama writers - this was part of a collaboration with PAWS an organisation that endeavours to improve the profile of science by encouraging Drama writers to include meaningful storylines

I have continued to interact with PAWS and some of the writers are working on storylines suggested as a result of their visit. A recent meeting involved a further visit from a Director and writer developing a drama based on the impact genetic diagnosis of susceptibility to breast cancer on a family. Current focus is on developing scripts and obtaining a commission from TV
Year(s) Of Engagement Activity 2007,2009
 
Description Inaugural Professorial Lecture 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Professorial lecture - open to the public and also attended by senior school pupils from local schools with an interest in biomedicine

Colleagues within the local scientific community were more aware of my research goals and interests and this has fostered collaborative discussions
Year(s) Of Engagement Activity 2007
 
Description International Competition for Sixth form students sponsored by MRC and Pfizer 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Schools
Results and Impact Interaction with school pupils - raised awareness of reproductive health.

Raised profile. Media coverage and good feedback from schools, MRC and Pfizer.
Year(s) Of Engagement Activity 2009,2010,2011,2012
 
Description Oration for Edinburgh Medal Lecture, Edinburgh Science Festival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Type Of Presentation Keynote/Invited Speaker
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Title of talk 'Oration for Edinburgh Medal Lecture by Prof Carl Djerassi

Raised my profile and public awareness of reproductive science
Year(s) Of Engagement Activity 2011
 
Description Women in Science Engineering and Technology 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? Yes
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact I was a member of a panel discussing the challenges faced by women scientists in their working lives - the discussion was chaired by Kirsty Walk and other panel members were drawn from media and training

This event had good press coverage
Year(s) Of Engagement Activity 2008