Perturbing the process of amyloid aggregation is a key strategy for the development of therapeutic agents in a range of neurodegenerative diseases. A variety of issues with current screening methods, including lack of reproducibility and high reagent consumption, can limit the efficiency of identifying small molecule inhibitors. This project aims to overcome such restrictions and identify new inhibitory molecules, through the development of a microfluidic based assay, which uses fluorescence lifetime imaging microscopy (FLIM) to monitor amyloid aggregation in up to 100 simultaneous assays. Aggregation of fluorescently labelled amyloidogenic monomers into larger structures is accompanied with a reduction in the fluorescence lifetime of the attached fluorophore dye, as a result of FRET processes. This phenomenon can be exploited to monitor the aggregation profile of such peptides and the effect that different extrinsic factors or inhibitory compounds can have on the process. The FLIM screening process has been coupled with droplet-on-demand technology and the development of a new microfluidic device, in which 100 precisely ordered droplets containing monomeric peptide together with test compounds can be imaged concurrently over several hours. Using this assay format, novel chemical libraries have been screened to identify potential inhibitors of the aggregation of amyloid β - the seminal neuropathic agent in Alzheimer’s disease. IC50 values of hit compounds have been calculated and have identified two molecular scaffolds capable of exerting a strong inhibitory effect on the aggregation process. In vivo tests, using adapted FLIM imaging techniques, are currently being developed to validate the potential therapeutic value of the hit compounds.

Identification and development of small molecule inhibitors of the aggregation of amyloid β

253rd National Meeting (2017)

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