Towards a novel 3D cell culture model to test the therapeutic efficacy of Affimers in the treatment of HER2 overexpressing breast cancer
Lead Research Organisation:
University of Leeds
Department Name: Sch of Molecular & Cellular Biology
Abstract
Breast cancer (BC) is the most commonly diagnosed cancer amongst women. Human epidermal receptor 2 (HER2) is overexpressed in 15-30 % of BC and is associated with poorer prognosis with comparison to the luminal subtypes. The monoclonal antibody (MAb) Trastuzumab is currently used for its treatment.
Mouse models have and continue to be valuable models of BC, however, to address the 3 R's it is desirable to reduce, refine and replace their use in research. Here, a novel in vitro 3D culture model of HER2+ BC is being investigated. The HER2 overexpressing cell line BT-474 were cultured as spheroids in a matrix-free medium, epiFL. Spheroids show characteristics of a tumour such as a necrotic core and 3D structure so are more translational to the clinic than 2D culture models. Cells were then cultured in SeedEZ scaffolds. The scaffolds are hydrophilic consisting of glass fibers that allow spheroids to form in between them. SeedEZ were cultured with PerfusionPal to more closely mimic the environment of a tumour in vivo. Furthermore Affimers, an antibody replacement protein scaffold, that bind HER2 have been identified and their effects on inhibiting HER2+ cell growth will be studied.
This is towards the creation of a fully humanised 3D model to test and compare the efficacy of Affimers in comparison to Trastuzumab in the treatment of HER2+ BC, potentially reducing the need for animals in preclinical drug testing.
Mouse models have and continue to be valuable models of BC, however, to address the 3 R's it is desirable to reduce, refine and replace their use in research. Here, a novel in vitro 3D culture model of HER2+ BC is being investigated. The HER2 overexpressing cell line BT-474 were cultured as spheroids in a matrix-free medium, epiFL. Spheroids show characteristics of a tumour such as a necrotic core and 3D structure so are more translational to the clinic than 2D culture models. Cells were then cultured in SeedEZ scaffolds. The scaffolds are hydrophilic consisting of glass fibers that allow spheroids to form in between them. SeedEZ were cultured with PerfusionPal to more closely mimic the environment of a tumour in vivo. Furthermore Affimers, an antibody replacement protein scaffold, that bind HER2 have been identified and their effects on inhibiting HER2+ cell growth will be studied.
This is towards the creation of a fully humanised 3D model to test and compare the efficacy of Affimers in comparison to Trastuzumab in the treatment of HER2+ BC, potentially reducing the need for animals in preclinical drug testing.
Organisations
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| NC/N00325X/1 | 01/10/2016 | 30/09/2019 | |||
| 2310402 | Studentship | NC/N00325X/1 | 01/10/2016 | 30/09/2019 | Sophie Roberts |