MICA: Control of tumour cell apoptosis by nuclear F-actin bundling

A large number of clinical studies have identified a protein called fascin that is present at extremely high levels in a range of human cancers. Very little is known about how this protein is controlled, but we know that reducing levels in cancer cells that we grow in the lab results in reducing the ability of those cells to undergo metastatic spread. Fascin is an exciting potential target to treat metastasis, but until we know more about how it is controlled, we cannot design drugs to target its function. This project aims to study the factors that control fascin function in cells and identify how the protein responds to changes in the environment that surrounds cancer cells. One of the major gaps in current knowledge is a detailed understanding of the factors regulating cancer progression and metastasis. This project will help to identify new proteins that can lead to increased cancer cell invasion and therefore provide new possible targets for designing future treatments. We will study the structure of the protein and identify specific the places and regions within this molecule that are important for controlling different functions in cells. We will then isolate proteins that bind to these forms of fascin and characterize how this binding occurs. We will then use human cancer cells in the lab to study these newly identified proteins that are responsible for promoting the fascin function and determine how they are controlled. Finally, we will use different strategies to inhibit these associated proteins and determine how they control cancer cell behavior that may lead to promotion of metastatic disease. Ultimately this study could lead to the identification of targets for the design and development of new drugs and therapeutic agents to prevent cancer growth and spread.

Fascin is an actin binding and bundling protein that is significantly up upregulated in most human cancers and contributes to metastasis. As fascin is not expressed in normal epithelial cells, it is a very attractive target for therapeutic intervention to halt tumour cell invasion. However, despite being discovered nearly 20 years ago and levels analysed in hundreds of cancers, we still know relatively little about the factors that control fascin function and this is a significant barrier to strategic development of therapies. Our recent published and unpublished data has revealed that fascin can associate with a number of previously undescribed binding partners and localizes to the cell periphery, nuclear envelope and inside the nucleus. Our exciting preliminary data has shown that nuclear localised fascin can regulate F-actin bundling within the nucleus, and when this phenotype is artificially enhanced, this results in commitment of tumour cells to apoptosis. The goal of the current study is to use a range of biochemical methods coupled with advanced cell-based imaging approaches to determine the molecular mechanisms that regulate nuclear fascin localisation under homeostatic conditions. In partnership with the major pharmaceutical company AstraZeneca, we will then employ a series of high-content, high resolution imaging screens to identify potential upstream regulators that can promote nuclear fascin and F-actin bundling leading to tumour cell death. These approaches combined will provide significant insight into novel mediators of fascin function and localisation, as well providing new tools and targets for modulators of nuclear F-actin. In the longer-term our goal is to use this information to develop new therapeutic strategies to target tumour cells and initiate cancer-specific cell death.

Academic:
We will present the data arising from this study at conferences in the UK, Europe and the US as well as through invited talks at Institutions. The current project also offers excellent multidisciplinary training opportunities for the PDRAs associated with this proposal. The PDRAs will be based within different labs (MP and SC) and very likely come from different training backgrounds (physics/computing and biology) but will work directly together for the duration of the project. The PI and co-I organise and teach on microscopy and image analysis workshops at King's, across the UK and internationally and will ensure the PDRA's join these meetings early on in the project to both extend their training and expand their networks in the field. The image analysis approaches to tackle high-content image screens being developed in this project will also be of great interest to many people within the imaging community and we plan to share these approaches at these workshops. We will also meet regularly with our industrial collaborators AstraZeneca throughout the project and inform them of progress from other aspects of the project, to provide the opportunity for us to receive a different perspective on the approaches we are using. This will also represent an important part of the PDRAs career development and provide excellent opportunities for exchange of ideas between academic and industrial team members.
Society:
We already conduct a number of outreach activities that we plan to extend to include concepts and data emerging from this proposal. We are organising a public microscopy/biophysics workshop at the Royal Microscopical Society (RMS) MMC meeting in 2019 meeting and will include examples of time-lapse movies, super-resolution data and images from the high content screening to demonstrate the power and importance of microscopy to the general public. We will use the data we acquire from live imaging experiments of fascin and actin in cells to explain how cells organise and change their architecture when they move, and how this is important in diseases, which in our experience always engages children very well. We will also engage yr12/13 school children at our bi-annual King's STARS (Science Training for Aspiring Research Scientists) week-long workshops run by MP. The STARS workshop offers the opportunity for schoolchildren to shadow scientists in the lab and we will ensure both PDRAs in the current project take on students in this programme to assist in their public engagement and outreach skills, as well as educating the next generation of researchers about our study and highlight how links between basic scientists and the pharmaceutical Industry are important in contributing to tackling diseases.
Industry:
The programme of research outlined here will analyse novel molecular events that co-ordinate cytoskeletal remodelling using advanced imaging approaches. Our experimental approaches and findings hold clear potential for the identification of drugs that target fascin and F-actin nuclear function in cancer. We will discuss our findings as they emerge with AstraZeneca and plan to progress any target hits to pre-clinical stage with additional funding either directly through AZ or via MRC DPFS applications or similar.
The importance of microscopy-based assays to analyse cell function is increasingly recognised as being extremely important in drug discovery by the pharmaceutical industry and the methods and reagents we plan to develop in this study will place us at the forefront of emerging technologies leading to the design and validation of new therapies. Our existing collaboration with AstraZeneca in this project means we are very well placed to explore future commercial potential arising from the current proposal. We believe our experience and expertise gained from this study will also place us in an excellent position to engage in other similar imaging development-based studies with other industrial partners.