This PhD will be a collaborative project, drawing on the expertise of Dr Andrew Fielding and Dr Morgan Gadd to develop a new small molecule to treat aggressive cancer types, in particular triple-negative breast cancer and metastatic uveal melanoma. It will be an interdisciplinary project, broadly encompassing the modern discipline of chemical biology, but specifically entailing synthetic chemistry, biochemistry, molecular biology, cell biology and pharmacology. Dr Fielding has worked extensively on the cell biology of cancer, whilst Dr Gadd’s research focuses on the construction and characterisation of cell-active degrader molecules.
Proteolysis-targeting chimaeras (PROTACs) are bifunctional degrader molecules that bring together a protein target and a ubiquitin-ligase enzyme. This leads to the target protein being “tagged” for degradation via the ubiquitin-proteasome system. PROTACs are recognised as potential drug-like molecules capable of degrading proteins on which cancer cells rely.
While normal cells contain two centrosomes, some cancer cells exhibit centrosomal amplification. This is a well-established phenotype in triple-negative breast cancer (TNBC). In addition, we have unpublished data showing it is also prevalent in metastatic uveal melanoma (MUM). Cells with centrosome amplification need to avoid multipolar mitosis-induced death by clustering their centrosomes into two functional groups, allowing for bipolar division. Centrosome clustering requires a specific kinesin, KIFC1. We have demonstrated that depleting KIFC1 using siRNA inhibits the growth of TNBC cells, yet does not affect non-cancerous breast cells. This shows that KIFC1 disruption can be a cancer-specific therapeutic approach.
In order to translate these findings towards the clinic, we now need a clinically applicable method to disrupt KIFC1 function. Two commercially available KIFC1 inhibitors have been shown to bind KIFC1. However, due to their pharmacokinetic properties, these inhibitors are unsuitable for clinical applications. Using these KIFC1-binding molecules as a starting point, the first strand of this project will use synthetic chemistry to develop PROTACs, as a strategy to degrade KIFC1 with small molecules.
The second, independent strand will investigate the reliance of Metastatic Uveal melanoma (MUM) cells on KIFC1 expression. MUM currently has particularly poor outcomes and therefore novel treatment approaches are urgently needed. Here, we will use cell growth assays and live-cell confocal microscopy of MUM cells with fluorescent cellular markers to test their reliance on KIFC1 function. We will also use live-cell imaging to investigate the normal dynamics of KIFC1 localisation and levels through the cell cycle.
The final section will use relevant cell culture models of TNBC and MUM to test the PROTAC. Firstly, the dynamics of KIFC1 degradation will be assessed by both western blotting and imaging of GFP-KIFC1. The effects on centrosome clustering will be assessed by immunofluoresence microscopy and, importantly, the downstream effects on cell growth of both cancer and matched normal cell lines will be determined.
Dr. Andrew Fielding and Dr. Morgan Gadd hold North-West Cancer Research Lecturer and Fellowship positions in the Division of Biomedical and Life Sciences at The University of Lancaster. Dr. Fielding’s interests lie in understanding unique features of cancer cells, and in turn targeting these to provide cancer-specific methods for inhibiting cell proliferation, progression and metastasis: http://www.research.lancs.ac.uk/portal/en/people/andrew-fielding(e70c4c4e-3d51-4aa6-bcdd-c08c39286735).html
Dr Morgan Gadd’s research interests are focused on the understanding and manipulation of protein-protein interactions and protein complexes, in particular how cancer driving and survival proteins can be therapeutically targeted through these associations: https://www.lancaster.ac.uk/bls/people/morgan-gadd
The Division of Biomedical and Life Sciences at The University of Lancaster (www.lancaster.ac.uk/bls/), was ranked no.5 in the UK for Biological Sciences by The Times/Sunday Times Good University Guide 2018. As well as the laboratory research project, PhD students in the division undergo a programme of research training activities, tailored to individual needs. Training opportunities include specific skills for postgraduate research students such as writing and presentation skills as well as a range of more generic courses such as teaching skills, statistical methods, computing and languages.
Applicants are invited from a range of chemistry/biochemistry/cell biology/cancer research backgrounds, although an enthusiastic approach to the proposed research area is essential. Laboratory experience covering any of the techniques mentioned above is desirable.
For informal enquiries, which are encouraged, please email Andrew Fielding at [email protected]
or Morgan Gadd at [email protected]
Applications are made by completing an application for PhD Biomedical and Life Sciences October 2019 through our online application system. Closing date: midnight 15th March 2019