Investigate the molecular mechanism controlling FEME (Fast Endophilin Mediated Endocytosis) in cancer cell migration using CRISPR, biochemistry, and advanced microscopy methods.
Not stated
- Funding
- Self-Funded PhD Students Only
- Application deadline
- Year-round applications
About the project
About the Project Cancer is a devastating disease: more than one in three people in the UK will develop cancer in their lifetime. Metastasis is the primary cause of cancer related deaths. Metastasis is caused by aberrant cell migration of cancer cells. Endocytosis is an essential process during development and tissue homeostasis ensuring diverse functions including modulation of growth factor signalling. Growth factor-receptor signalling such as the epidermal growth factor receptor (EGFR) controls proliferation as well as directed migration including cancer cell migration. We have shown that the protein Lamellipodin (Lpd) cooperates with the protein endophilin to promote clathrin-mediated EGFR endocytosis (Vehlow et al., EMBO J. 2013) and cancer cell migration through the actin regulatory Ena/VASP proteins and the Scar/WAVE complex (Law et al., Journal of Cell Biology, 2013; Carmona et al., Oncogene, 2016). In addition, Lpd recruits endophilin to the leading edge of cells thereby inducing clathrin-independent endocytosis ( F ast, E ndophilin- M ediated E ndocytosis (FEME)) and some regulations of FEME have been explored (Boucrot et al., Nature, 2015; Wah-Hak et al., Nature cell Biology, 2018; Casamento and Boucrot, Biochem. J., 2020). However, we still do not understand the fundamental mechanism of FEME. Actin polymerisation is essential for FEME and may provide the force for invagination and scission, but we do not how actin polymerisation is controlled to mediate FEME and its integration with membrane bending BAR domain containing proteins. In this project, which will start in October 2026, you will investigate the molecular mechanisms of how actin polymerisation contributes to FEME. Our hypothesis is that this is mediated by Lpd, Ena/VASP proteins, the Scar/WAVE complex, and additional actin regulators. In addition to Lpd, we already identified another key protein, NHSL1 mediating FEME which links to actin regulators. You will generate CRISPR-knockout cell lines and rescue them with cDNA mutated in the binding sites. You will characterize resultant cell lines by super-resolution live cell microscopy methods for defects/efficiency in FEME. You will also generate mStaygold/mScarlet knock-in cell lines for evaluating timing of FEME using TIRF and/or super-resolution live cell imaging. You will use biochemical ELISA-based endocytosis assays to quantify EGFR uptake. Taken together, your PhD work will unravel a novel and general control mechanism of FEME endocytosis in migrating cancer cells. You will join a friendly, interactive lab, which is part of the Cellular Biophysics Section of the Randall Centre at King’s College London: 11 laboratories with shared interest in the regulation of the cytoskeleton in cell division, adhesion, migration, and intracellular trafficking with joint meetings. Furthermore, our lab is part of the UK wide UK Cell Motility Club which I am organising.