Paracrine signalling pathways in endometriosis and related cancers

Endometriosis, the presence of endometrial-like cells (glands and stroma) outside the uterus, is a common debilitating condition in women of reproductive age. Similar to the womb lining itself, these endometriotic cells are fully responsive to female sex steroids and undergo repeated episodes of bleeding at the onset of menstruation. Release of cell debris, erythrocytes, haem-bound iron, cytokines and other pro-inflammatory mediators into ovarian endometriomas or the peritoneal cavity support endometriotic cell growth and survival at lesion sites. The presence of redox active iron has also been shown to induce oxidative stress, which can contribute to disease progression and risk of carcinogenesis. Understanding these aberrant pathways is crucial to exploit the metabolic activity of endometriotic cells and successfully treat this enigmatic disease.

We are very interested in the epidemiological link between endometriosis and cancer and aim to be the first to model this malignant transformation and evaluate preventative measures by:
- Identifying the lipid signature and metabolism of endometrial (control), endometriotic and cancer cells.
- Modelling the in situ environment.
- Assessing the in vitro effects of promising new therapeutic agents.

Endometrial and adipose tissues will be collected from consenting women of known provenance undergoing routine gynaecological surgery at St Mary's hospital, Manchester. Primary cells will be cultured in vitro and the metabolic capacity of control, endometriotic and endometrial cancer cells alone in co-culture with adipocytes will be assessed under normoxic and hypoxic conditions using the XF flux (Seahorse) analyser, a state-of-the-art redox system. Other measurements will include: cell viability (trypan blue exclusion/ MTT), proliferation (Ki-67/ apoptosis markers (ICC, flow cytometry)) steroidogenesis (qRT-PCR, ELISA) and transwell migration and invasion (fluorescence microscopy). The effect of novel therapeutic agents on cell glycolysis will also be investigated as well as the release of inflammatory mediators using lipidomic and Luminex technologies. Findings will be correlated with clinical history and patient-reported symptoms to stratify data for personalised medicine.

Understanding the nature of the unique local environment should provide new insights into disease pathogenesis and progression as well as expedite drug design and delivery.