Project ID CM-HD2023_12

ThemeCM-HD

Co Supervisor 1A Randall Division/ChemistryWebsite

Co Supervisor 1B Physics/Randall DivisionWebsite

The effect of lipid composition on the mechanostransduction of individual live cells

Are the lipids forming the plasma membrane, organelles and nuclear envelope (NE) dynamically modified under mechanical stress? Lipids and proteins are key components of membranes, yet most of the effort to understand mechanotransduction has focused on proteins alone. First, we will explore whether the lipidome changes in the plasma membranes, organelles and NE of cells exposed to mechanical stress. We will subject cultured cells to substrates of different stiffness, and extract their nuclei. Plasma membrane and nuclear lipids will be extracted and analysed by MS to determine their lipidomic profiles. In parallel, we will use Atomic Force Microscopy (AFM) in combination with magnetic tweezers cell stretching experiments to probe the mechanical properties of plasma and nuclear membranes.

We will investigate the effect of mechanical forces on the lipid composition of cells and isolated nuclei and organelles. The student will gain expertise in single cell AFM and magnetic tweezers characterisation, combined with cell and molecular biology techniques. S/he will also gain deep knowledge in mass spectrometry. In Year 1, cell biology experiments will be performed at UE lab and the student will learn how to prepare substrates of different stiffness in SGM lab. Year 2 will be devoted to conduct single cell mechanical experiments using AFM and Magnetic Tweezers (SGM). During Year 3 the student will concentrate on lipidomics (UE). Experiments, analysis and paper writing will continue in Year 3-4.

This is a unique opportunity to explore fundamental biophysical questions of lipids during mechanotransduction at the single cell level, combining cutting-edge nanomechanical biophysical techniques (Garcia-Manyes) and modern cell biology and mass spectrometry (Eggert).

One representative publication from each co-supervisor:

• Atilla-Gokcumen, Muro, E.; Relat-Goberna, J.; Sasse, S.; Bedigian, S.; Coughlin, M.L.; Garcia-Manyes, S.; Eggert, U.S.; «Dividing cells regulate their lipid composition and localization» Cell (2014), 156 (3), 428

• Infante, E*.; Stannard, A.*; Board, S.J.; Rico-Lastres, P.; Rostkova, E.; Beedle, A.E.M.; Lezamiz, A.; Wang, Y.J.; Gulaidi Breen, S.; Panagaki, F.; Sundar Rajan, V.; Shanahan, C.; Roca-Cusachs, P.; Garcia-Manyes, S. The mechanical stability of proteins regulates their translocation rate into the cell nucleus. Nature Physics, 2019