Investigate the molecular mechanism controlling FEME (Fast Endophilin Mediated Endocytosis) in directed cancer cell migration using CRISPR, biochemistry, and advanced microscopy methods.

King’s College London

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 (Fast, Endophilin-Mediated Endocytosis (FEME)) (Boucrot et al., Nature, 2015; Wah-Hak et al., Nature cell Biology, 2018; Casamento and Boucrot, Biochem. J., 2020). We identified another protein interacting with Lpd which may control certain steps in FEME (unpublished).

In this project, which will start in January/June/October 2024, 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 and the Scar/WAVE complex. You will also explore how Lpd together with the other identified protein mediates EGF-dependent directed cancer migration (chemotaxis) by controlling FEME and recycling of the EGFR to the leading edge. 

You will generate CRISPR-knockout cell lines and rescue them with cDNA mutated in the binding sites. You will also generate mGreenlantern/mScarlet knock-in cell lines. You will characterize resultant cell lines by advanced live cell microscopy methods for defects/efficiency in FEME. You will use biochemical ELISA-based endocytosis assays to quantify EGFR uptake. For chemotaxis analysis you will use a well-established microfluidic chamber.

Taken together, your PhD work will unravel a novel and general control mechanism of FEME endocytosis supporting cancer cell migration.

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 London wide London Cell Motility Club.

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