We recently developed compounds aimed at disrupting the interactions between CD44 and FERM proteins; an interaction believed to be pathological in Alzheimer?s disease (AD). We developed small molecules: 1) using in silico docking at the site where CD44 would bind to FERM proteins, 2) Validated the binding using microscale thermophoresis (MST) and surface plasmon resonance (SPR) and 3) validated disruption of CD44 peptide binding to FERM proteins using MST. We are now developing assays to validate binding in the cell using cellular thermal shift assay (CETSA) and fluorescently tagged CD44 to define the disruption between CD44 and FERM proteins. We are also starting to implement assays to define compounds that have a potential to be neuroprotective. Since these proteins were found to be upregulated in microglia, we are developing flow assays to measure the effect of compounds for microglia-specific functions such as oxidative stress and phagocytosis. These functions will be helpful in further developing the compounds for future development of therapeutics for AD.
The research proposed in this supplement is aimed at defining the mechanism of action of compounds developed that target CD44/FERM interaction for AD therapeutics. We are developing tools and defining their action using CETSA (to measure target engagement), Fluorescently tagged protein to define their disruption in cells and cell-based assays to define their effects in microglia.
