Investigate the role of NHSL1 in vesicle trafficking and receptor recycling promoting cancer cell invasion using CRISPR-Cas9, microfluidics, and advanced live cell imaging.

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. We have identified NHSL1 as a key regulator of cell migration (Law et al., Nature Communications, 2021)¹. NHSL1 is part of the poorly characterized Nance-Horan syndrome protein family. We showed that NHSL1 negatively regulates cell migration via the Scar/WAVE-Arp2/3 complexes which control actin filament nucleation required for cell migration¹. Mechanistically, NHSL1 inhibits Scar/WAVE-Arp2/3 activity and consequently lamellipodia stability and cell migration efficiency¹. In addition, we observed that NHSL1 localises to vesicles which emanate from the leading edge of migrating cancer cells suggesting that it is involved in vesicle trafficking¹. NHSL1 interacts with additional actin effectors (unpublished). During guided migration, growth factor receptors are endocytosed at the leading edge and traffic back there to increase polarity and directional migration. However, the molecular details of the pathways controlling this are still enigmatic.

In this project, which will start in October 2024, you will investigate the role of NHSL1 in growth factor receptor recycling supporting directional migration of cancer cells.

You will evaluate how NHSL1 interacts with additional actin effectors. You will use biochemistry to map and Gibson assembly to mutate the binding sites of these additional actin effectors. You will generate CRISPR-knockout cell lines and rescue them and existing NHSL1 CRISPR KO cell lines with cDNA mutated in the binding sites. You will utilise advanced imaging to track and quantify changes in the trafficking of the receptors. You will evaluate the functional significance of these interaction for chemotaxis of cancer cells using a well-established microfluidic chamber and using 3D invasion assays.

Taken together, your PhD work will unravel a novel control mechanism of receptor recycling 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.