Project ID CM-HD2023_23

ThemeCM-HD

Co Supervisor 1A Peter Gorer Deparment for Immunobiology/SIMSWebsite

Co Supervisor 1B Comprehensive Cancer centreWebsite

The role of the actin cytoskeleton for cytotoxic T cells

Our group is interested in the understanding the signalling pathways that control the migration of cytotoxic T cells (CTLs) into tumour tissues and the subsequent killing of their target cells. We do this by a combination of in vitro biochemistry and cell biology approaches, including CRISPR knockouts, advanced microscopy techniques and in vivo mouse model approaches. Our lab has recently identified the kinase WNK1 as a novel and important regulator of T cell killing and actin dynamics. In the absence of WNK1, CTLs fail to migrate into tumours and do not properly rearrange their actin cytoskeleton upon binding to a target cell. As a consequence, they are strongly defective in killing.

In this project the student will use primary mouse T cells to study the role of WNK1 in controlling TCR induced actin remodelling. During the first year research will focus on imaging actin dynamics during T cell killing in vitro through advanced microscopy techniques to further characterize the phenotype. From the second year onwards, the student will use biochemistry approaches to investigate the molecular basis of WNK1-signalling, concentrating on pathways that control actin polymerization and acto-myosin contractility based on preliminary data from lab. This will be done through immuno-blotting experiments, CRISPR-knockouts and overexpression studies to identify WNK1-dependent pathways and from year three onwards through pull downs, interaction studies and phosphoproteomics approaches to identify how WNK1 activates those pathways.

One representative publication from each co-supervisor:

• WNK1 kinase balances T cell adhesion versus migration in vivo. Köchl R, Thelen F, Vanes L, Brazão TF, Fountain K, Xie J, Huang CL, Lyck R, Stein JV, Tybulewicz VL. Nat Immunol. 2016 Sep;17(9):1075-83.

• Invadopodia play a role in prostate cancer progression (2022) Valeria Manuelli , Fidelma Cahill , Harriet Wylie , Cheryl Gillett , Isabel Correa , Susanne Heck , Alex Rimmer , Anna Haire , Mieke Van Hemelrijck, Sarah Rudman and Claire M. Wells BMC Cancer (2022) 22:386 https://doi.org/10.1186/s12885-022-09424-4 RESEARCH