Deciphering the Molecular Dynamics of Cell Adhesion and the Role of Biomechanics in Ehlers-Danlos Syndrome

The connections a cell forms with its environment underlies one of the most fundamental processes of organisms, with aberrant connections leading to the development of pathological conditions. Integrins are essential for mediating these connections to the surrounding Extracellular Matrix (ECM) through formation of adhesion sites which are crucial for tissue integrity. Depending on pathological condition, the expression level of integrin isoforms change, which may be occurring in the connective tissue disorder Ehlers-Danlos Syndrome (EDS). EDS cells display a reduced expression of the fibronectin receptor integrin with the consequent recruitment of the alternative receptor integrin. However, it is also assumed the molecular composition of these adhesion sites also sets their strength and stability, ultimately influencing the actin cytoskeleton and a cell's biomechanical properties which has not been studied in EDS. The mechanochemical details of the formation of these adhesion sites also remain poorly understood. We propose using a newly developed assay that employs functionalised elastomers to mimic the ECM, allowing control of the mechanical and biochemical environment into which cells are cultured. This allows to study the role of adhesion protein type and mixture in adhesion site formation. Combination of the elastomer to a uniaxial cell-stretcher also allows for stresses to be applied during cell adhesion. Live cell microscopy combined with image analysis tools to quantify imaging data will then provide deeper insights into the dynamics of cell adhesion. Through this, we seek to understand how an altered integrin profile can affect the mechanical properties of the cell, how adhesion site and actomyosin dynamics change in response to mechanical stimuli, and if this is affected in EDS. The results of this project will provide major advances in our understanding of the processes involved in maintaining tissue integrity in conditions like EDS.