Investigating the role of Ect2 in pancreatic cancer cell proliferation and motility

About the Project

Pancreatic ductal adenocarcinoma (PDAC) has a low survival rate and new therapies are urgently needed. The RhoGEF, Ect2, is frequently overexpressed in PDAC tumours and is associated with a poor prognosis. Ect2 modulates the contractility of the actin cytoskeleton by locally activating RhoA and plays a role in multiple cellular processes including cell division, DNA repair and cell migration. However, it’s not clear which of these roles are most important for its function in PDAC tumorigenesis. In addition, Ect2 is regulated by the YAP/TAZ mechano-sensitive signalling pathway and our preliminary data indicate that its subcellular localisation is altered when cells are grown on stiffer substrates. This is likely to have implications for proliferation of cancer cells within PDAC tumours, which are mechanically extremely stiff. This project will investigate the role of Ect2 mechano-sensing in pancreatic cancer cells and patient tumours.

Specifically, this project aims to:

1. Characterise the effect of Ect2 depletion on PDAC cell proliferation, division and migration
2. Understand how mechanosensitive signalling regulates Ect2 localisation and activity
3. Image the localisation of Ect2 in PDAC patient tumours

The project will use a combination of cell and molecular biology approaches (including CRISPR gene editing, western blotting, flow cytometry), advanced microscopy (live cell imaging & immunofluorescence) and histopathological staining of patient samples to elucidate the role of Ect2 in PDAC. To investigate the mechano-regulation of Ect2, cells will be grown on soft and stiff hydrogels to mimic the increasing tissue rigidity within PDAC tumours. Ultimately, this project aims to develop Ect2 as a potential new target for PDAC therapy.

Lab website:

https://www.helenmatthewslab.org/

Please apply for this project using this link: https://www.sheffield.ac.uk/postgraduate/phd/apply/applying

Funding Notes

Externally or self-funded students only