A 3D tissue model of breast morphogenesis for replacing animals in testing for endocrine disrupting substances

Endocrine disrupting substances (EDS) are environmental chemicals that alter endocrine system function. The most striking deleterious effects have been reported after EDS exposure during embryonic and fetal life. We have previously developed screens to identify estrogen and androgen agonists and antagonists using cell cultures. These assays are efficient in determining hormonal activity, but they cannot ascertain the occurrence of a deleterious effect in tissues. Unfortunately the only reliable way at present to test for the developmental effects of EDS is to use animals for research that are sacrificed at different development stages. We therefore propose to develop a robust and reliable model of breast tissue, using estrogen-sensitive human cell lines that would mimic the normal architecture of the breast and will contain both epithelium and stroma. We will build on the previous 3D model that we have developed which revealed the biomechanical conditions needed for duct formation. By including epithelial estrogen-responsive cells, we will be able to mimic the normal action of estrogens as well as the pathological outcomes observed after exposure to diethyl stilbestrol (DES) and bisphenol A (BPA), which include intraductal hyperplasias and stromal desmoplastic lesions. This work will require a multidisciplinary scientific approach combining skills in tissue engineering, cell culture, microscopy and measurement science. This will be provided for by the joint experience of the collaborators and a new tissue engineering laboratory built around a Leica TCS STED microscope, as well as the existing advanced electron microscopy facility. Professor Howard is a medically qualified pathologist who has many years experience in regulatory toxicology and stereology. Professor Ana Soto is a medically qualified cell biologist who has developed the current knowledge-base in 3D breast tissue culture as well as the E-screen and A-screen tests. Dr George McKerr is the Director of the FEI Centre for Advanced Imaging and is a widely experienced biologist and microscopist. Dr Kurt Saetzler has expertise in 3D reconstruction and digital image processing. We anticipate that the end result will lead to an exportable testing system that enables ourselves and others to reliably identify harmful doses of EDSs without the need to use animals and, at the same time, allowing us to directly observe the damaging process caused by EDS. This will potentially give further clues on how EDSs work in real life.