Alzheimer’s disease (AD) is a devastating neurological disorder characterized by the deposition of aggregated proteins in the brain in the form of extracellular beta-amyloid in senile plaques and intracellular tau in neurofibrillary tangles. A current approach towards treatment of Alzheimer’s disease is by using inhibitors of amyloid beta aggregation. Current screening protocols for inhibitors of amyloid beta aggregation generally involve first using assays for fibers, e.g, thioflavin T binding to detect inhibition of fiber formation or fiber disassembly, and the compounds uncovered in these screens are then examined for effects on oligomer formation. This approach for screening compounds having amyloid beta aggregation inhibition properties is an indirect one, but is used because oligomers do not bind thioflavin T and there is no easy assay for their appearance. A compound which only inhibits oligomer formation, but does not inhibit protective fiber growth would never be detected by this indirect approach. The present UIC invention overcomes these deficiencies and provides an easy and direct assay in yeast for high-throughput screening of compounds that can inhibit amyloid-beta oligomer formation but does not inhibit protective fiber growth.
Description/Details

The present UIC invention is directed towards a yeast high-throughput screen for detecting compounds that inhibit amyloid-beta aggregation. It also provides a yeast in vivo assay for amyloid-beta aggregation. The assay involves replacing the N-terminus of the translational release factor, Sup35, with Abeta-42mer, and examining the activity of said construct in an ade1-14 yeast strain in which the normal Sup35 gene was deleted, and inhibition of release factor translation termination activity of the fusion construct can be assayed for growth on –Ade medium.
Applications

Screening for compounds that inhibit amyloid-beta oligomer formation
Benefits

• Easily detects compounds that inhibit amyloid-beta oligomer formation but do not inhibit protective fiber growth Selects compounds with more “drug-like” properties (e.g., membrane permeability and cytotoxicity effects) compared to biochemical HTS screens, Clean read-out against a null background in a heterologous, yet eukaryotic environment, compared to mammalian cells, Self-renewal system, Simple handling, Fast discrimination of real hits from false positives, Inexpensive culture conditions
source: otm.illinois.edu