Lab rotation project description
The project will enable the initial evaluation and optimisation of 3D culture of brain tumour cells in peptide hydrogels. Using methods already proven for stem cells and breast tumour cells we will compare the response of cells in ‘naked’ non-functionalised gels with those incorporating common matrix proteins such as collagen IV, laminin and perlecan. Glycans (heparan and chondroitin sulphate) will also be tested. The gels can additionally be tailored with regard to their stiffness, a characteristic known to influence cell response. Culture of medulloblastoma, ependymoma and glioma cell lines is well established in the Coyle group and cells will be characterised for their proliferation, morphology and migration within the hydrogels. Stromal cells will be incorporated to establish co-culture. Where possible, live imaging using vital dyes will be used to detail cell interactions. These experiments will enable the identification of optimal seeding densities and growth protocols for the tumour cells and will additionally provide initial evidence for differential matrix dependency between the cell types.
Industrial biotechnology and bioenergy
Centre for Biomolecular Sciences / Queen's Medical Centre
LR1, LR2, LR3
Anna.Grabowska@nottingham.ac.uk / Beth.Coyle@nottingham.ac.uk
There remains a critical need for improved provision of in vitro test environments enabling the tissue-realistic culture of cells for mechanistic study as well as drug and toxicity screening.
Given the extensive cost of developing novel tumour therapeutics and the known importance of 3D growth on tumour cell behaviour, in vitro tumour modelling is an area of particular academic and commercial interest. The matrix surrounding cancer cells plays an important role in disease progression, with reciprocal signalling enabling cells to ‘read’ messages from the matrix and for tumour cells to re-engineer their matrix environment to influence neighboring cells.
Currently, modeling of tumour-matrix interactions uses animals (typically rodents) or commercial matrix preparations in 2D culture in the lab. Neither option accurately replicates the complex protein and sugar compositions of human matrix and they fail to reflect the tissue-specific differences fundamental to tumour growth. We have specialist expertise in proteoglycans, proteins decorated with glycans that have been implicated in many cancers, including those in the brain.
Recently, we developed a fully synthetic, highly reproducible hydrogel based on a simple short peptide motif. Cancer cells, along with other cell types, can be encapsulated in the gels and easily grown in the lab. A novel attribute is the ease with which the gels can be functionalized with relevant proteins/peptides or glycans to create bespoke matrix environments, to mimic the tissues in which tumours develop. The hydrogels have been optimized for reproducibility and stability, both characteristics often problematic for in vitro matrices and the gelator peptide is small and relatively cheap to manufacture making scale-up of the technology economically viable.
In this highly interdisciplinary project linking cancer cell biology, biomaterials design and biochemistry, we aim to generate a panel of hydrogels that mimic normal and tumour matrix environments in the brain. We will conduct linked proteomic and glycomic analysis of brain tissue sections (normal and medulloblastoma, ependymoma and glioma) to identify distinctive matrix components characteristic of each tissue type. These will then inform the proteins and glycans selected to functionalise bespoke hydrogels tuned to match the tissues. Tumour cells, as well as other stromal cells, will be encapsulated into the gels and their behaviour (proliferation, morphology, invasion) compared between the various matrices and with current in vitro and in vivo models. This project will run alongside a complementary study funded by NC3Rs using a similar approach to create hydrogels to mimic dense and non-dense breast environments to model breast cancer.
There is a real opportunity for added value here with the potential to use the brain gels to evaluate metastatic spread of breast cancer and to improve our understanding of how distinct matrix environments in different tissues support tumour growth and how this can be used to design improved therapeutic strategies. The NC3Rs project additionally provides access to low- or zero-cost matrix proteins and glycans that can be used to functionalise the hydrogels.
The project addresses the key BBSRC themes of tissue engineering and cell-cell communication and will provide much-needed interdisciplinary training to prepare the researcher for a career in modern bioscience.