A 3-D tissue model of breast morphogenesis for replacing animals in testing for endocrine disrupting substances
Lead Research Organisation:
University of Ulster
Department Name: Research Office
Abstract
Hormones are natural chemicals which carry messages to cells in the body from the brain or various (endocrine) glands. Endocrine disrupting substances (EDS) interfere with the passing of messages by hormones. Development of babies in their mothers? wombs is very dependent on the correct very low level of hormone. Any EDS present may corrupt the correct developmental signal being sent to the cells of the baby and cause problems. The Endocrine Society in 2009 published a statement on endocrine disrupting substances (EDS) invoking the Precautionary Principle.
The only reliable way at present to test for the effects of EDS on development in the womb is to use animals. Because it is very difficult to perform direct measurements on living embryos, studies have to sacrifice animals at different stages of development.
We wish to develop a model of breast tissue, using human cells from commercially available cell lines, so that we can test EDS and see if they affect development. This is building on results from our previous research based on 2D cell cultures detecting whether a chemical is an EDS that mimics the female and/or male sex hormones. This has already replaced many animal experiments. However, to detect the damage that can be caused by EDS during development we need to establish a new type of cell culture model that allows us to engineer, control and observe a model tissue consisting of the different types of cells found in the real tissue during life and exhibiting the same intriguing complex ductal branching structure.
This task will require a cooperative effort by scientists with skills in tissue culture, microscope usage and measurement science. Our University in Ulster has just invested #654k in a new microscope and tissue engineering facility that will help us to achieve this task. The internationally renowned company, Leica, who developed and provides this equipment, expressed their interest in supporting and helping us in this task. We are therefore asking the NC3Rs for funds to find skilled manpower and consumable items.
We anticipate that the end result will lead to a testing system that enables us to reliably identify harmful doses of EDSs without the need to use animals and at the same time allow us to directly observe the damaging process caused by EDS. This will potentially give further clues on how EDSs work in real life.
The only reliable way at present to test for the effects of EDS on development in the womb is to use animals. Because it is very difficult to perform direct measurements on living embryos, studies have to sacrifice animals at different stages of development.
We wish to develop a model of breast tissue, using human cells from commercially available cell lines, so that we can test EDS and see if they affect development. This is building on results from our previous research based on 2D cell cultures detecting whether a chemical is an EDS that mimics the female and/or male sex hormones. This has already replaced many animal experiments. However, to detect the damage that can be caused by EDS during development we need to establish a new type of cell culture model that allows us to engineer, control and observe a model tissue consisting of the different types of cells found in the real tissue during life and exhibiting the same intriguing complex ductal branching structure.
This task will require a cooperative effort by scientists with skills in tissue culture, microscope usage and measurement science. Our University in Ulster has just invested #654k in a new microscope and tissue engineering facility that will help us to achieve this task. The internationally renowned company, Leica, who developed and provides this equipment, expressed their interest in supporting and helping us in this task. We are therefore asking the NC3Rs for funds to find skilled manpower and consumable items.
We anticipate that the end result will lead to a testing system that enables us to reliably identify harmful doses of EDSs without the need to use animals and at the same time allow us to directly observe the damaging process caused by EDS. This will potentially give further clues on how EDSs work in real life.
Technical Summary
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, (which the University of Ulster has recently purchased for #654k), as well as the existing advanced electron microscopy facility. Professor Howard (PI) 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.
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, (which the University of Ulster has recently purchased for #654k), as well as the existing advanced electron microscopy facility. Professor Howard (PI) 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.