The Role of Sphingosine-1-Phosphate in Endothelial Integrity (360G-Wellcome-211504_Z_18_Z)

Sphingosine-1-phosphate (S1P) has been well established as a signalling lipid involved in the regulation of vascular permeability via binding of one of five G protein coupled receptors called S1PR1-S1PR5. This project aims to develop a model for measuring S1P control of vascular permeability, in order to demonstrate the usefulness of S1P as a therapeutic for reducing vascular permeability during organ transplantation. Previous work by this group have shown that both Human Microvascular Endothelial Cells (HMEC-1), and a positive control (Caco-2), grown to confluency on 0.4um transwells present a barrier to Evans Blue dye conjugated to albumin. This model will be manipulated by treating these cells with either S1P or agonists/antagonists of the different S1P receptors to show their effect on the ability of HMEC-1 to form a barrier to Evans Blue. This work will be supported by measuring the changes in electrical resistance across the endothelial barrier on these transwells following the aforementioned treatments, using an ohmmeter. Confluency of both cell types will be demonstrated using H & E staining. Together these results will demonstrate the ability of S1P to control endothelial permeability in vitro via the differing S1P receptors and therefore its potential for use as a therapeutic in organ transplantation.