Project ID CM-HD2023_25

ThemeCM-HD

Co Supervisor 1A Randall Centre for Cell and Molecular Biophysics, FOLSMWebsite

Co Supervisor 1B School of Cancer and Pharmaceutical SciencesWebsite

Nanoscale regulation of actin nucleation at the leading edge of migrating cancer cells.

Most cancer patients die from metastasis, the dissemination of the primary tumour after cancer cells acquired the ability to migrate. Mesenchymal cancer cell migration is driven by actin filament nucleation at the leading edge of cells, the lamellipodium. They are nucleated into a dendritic array by the Arp2/3 complex which is activated by Scar/WAVE. Lamellipodia rapidly protrude and retract in an oscillatory manner to explore the extracellular matrix controlled by key regulators of Scar/WAVE, NHSL1 and Lamellipodin, which we have identified acting in feedback and feedforward loops. It is unknown how the protein complexes forming these signalling circuits dynamically interact at the nanoscale to form a narrow actin nucleation zone at the very edge of lamellipodia.

You will join our dynamic research teams, which are long-term collaborators. With expertise and supervision from the Ameer-Beg lab, you will modify a unique, existing, fast, live cell fluorescence lifetime microscope (FLIM) (a 1024 beam multifocal FLIM with 40Hz frame rate for single-molecule localisation and image correlation) and develop data analysis to perform and quantify single-molecule localisation FLIM.

With expertise and supervision from the Krause lab you will tune current NHSL1, Lamellipodin probes, and our ARP2/3 complex biosensors and develop novel Scar/WAVE biosensors for application in single molecule FLIM for use on this microscope.

Your work will, for the first time, allow the elucidation of the complex lamellipodia feedforward and feedback networks.

In summary, you will contribute to a detailed understanding of nanoscale signalling circuits controlling lamellipodia dynamics driving cancer cell migration.

Representative Publications

• Law, A.-L., Jalal, S., Pallett, T., Mosis, M., Guni, A., Brayford, S., Yolland, L., Marcotti, S., Levitt. J.A., Poland, S.P., Rowe-Sampson, M., Jandke, A., Köchl, R., Pula, G., Ameer-Beg, S.M., Stramer, B.M., and Krause, M. (2021) Nance-Horan Syndrome-like 1 protein negatively regulates Scar/WAVE-Arp2/3 activity and inhibits lamellipodia stability and cell migration. Nature Communications, 12(1): 5687; DOI: 10.1038/s41467-021-25916-6

• Levitt, J. A., Poland, S. P., Krstajic, N., Pfisterer, K., Erdogan, A., Barber, P.R., Parsons, M., Henderson, R. K., Ameer-Beg, S.M., Quantitative real-time imaging of intracellular FRET biosensor dynamics using rapid multi-beam confocal FLIM. Sci Rep 10, 5146 (2020).