Cancer is a devastating disease: more than one in three people in the UK will develop cancer in their lifetime. 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 Arp2/3 complex-mediated actin filament nucleation at the leading edge of cells.
We have evidence that the novel NHSL1-A complex interacts with the Arp2/3 complex via its WCA domain and controls mesenchymal cell migration efficiency and chemotaxis (Jalal et al., 2025). However, we do not know how the NHSL1-A complex functions and how it is controlling cancer cell migration and invasion in 3D in physiological tumour microenvironment settings required for metastasis.
In this interdisciplinary project during the first year, you will use biochemistry to in vitro reconstitute the NHSL1-A complex from purified proteins and investigate how it activates the Arp2/3 complex through its WCA domain and the regulation thereof by GSK3 and CK2 (Aim 1).
In parallel in year 1-2 you will use CRISPR to specifically knock-out the NHSL1-A isoform required for formation of the NHSL1-A complex and create dominant-negative and active versions of NHSL1-A in the WCA domain by CRISPR mediated knock-in into the NHSL1 gene in suitable cancer cell lines with high expression of the NHSL1-A isoform (Aim 2). You will also generate fluorescent protein (mStayGold) tagged NHSL1-A cell lines by CRISPR-Cas9 knock-in and evaluate them using live cell microscopy (rotation).
In year 2-3 you will use the mStayGold-NHSL1-A cell lines in advanced live cell microscopy experiments to quantify Arp2/3 activities using FRET-FLIM biosensors and correlate these with leading-edge speed and F-actin assembly rates. You will also use advanced live cell imaging to evaluate the function of the NHSL1-A complex and its role in activating the Arp2/3 complex controlling cancer cell migration and invasion in 3D in physiological tumour microenvironment settings required for metastasis using spheroids generated from above cancer cell lines (Aim 3).
At the end of year 3 you will write and submit a manuscript and in year 4 you will do the revision work required for publication and write up and defend your thesis.
You will acquire a wide range of skills including biochemical assays, cloning, CRISPR knockout and knock-in, 3D spheroid invasion assays, physiological stiffness hydrogels, advanced live cell microscopy (super-resolution and FRET-FLIM).
You will join two friendly, interactive labs in the Randall Centre at King’s College London.
You will also generate fluorescent protein (mStayGold) tagged NHSL1-A cell lines by CRISPR-Cas9 knock-in and evaluate them using live cell microscopy (rotation).”