A microfluidic fluorescence lifetime screening platform bridges in vitro and in vivo formats and enables discovery of Amyloid β aggregation inhibitors

Inhibiting the aggregation of Amyloid β (1-42) is a promising strategy for the development of disease-modifying Alzheimer’s disease therapeutics. To date, however, no sufficiently efficacious inhibitors have been identified, despite the best efforts of more than 200 advanced drug development campaigns. This failure can be partially attributed to limitations in current in vitro compound screening and in vivo validation assays. Here, we report a high throughput assay platform based on the use of a fluorescence lifetime aggregation sensor that can be applied in vitro and in vivo. This microfluidic “nanoFLIM” assay presents a cost-effective and reliable alternative to conventional early-stage small molecule discovery platforms for cell-free Amyloid β aggregation. The fluorescence lifetime sensor can dynamically monitor peptide aggregation in cellular and Caenorhabditis elegans disease models, providing directly comparable aggregation kinetics, which is not achievable by any other method. The power of this unified platform for accelerating hit-to-lead strategies, lowering attrition rates and expediting in vivo screening, was demonstrated with a pilot screening campaign of 445 compounds, revealing a new inhibitor that can inhibit Amyloid β self-assembly both in vitro as well as in cellular and whole organism disease models.