High-Throughput Analysis of Amyloid Deposition

Park, Chan

Abstract

High-Throughput Analysis of Amyloid Deposition The formation of amyloid deposits in tissue is a hallmark of many pathologies such as Alzheimer's, Parkinson's, type II diabetes, mad cow, and other life-threatening diseases. Despite many studies on amyloid over a century, a clear understanding on how normal and soluble proteins assemble into fibrillar and insoluble amyloid aggregates under certain conditions remains unresolved. The key to unraveling the underlying etiology of amyloid-related diseases lies in the design of controlled in vitro experiments that can allow amyloid deposition study under selected environmental conditions that are known to induce normal protein to undergo aggregation. Amyloid deposition, the process of amyloid growth by the association of individual soluble amyloid molecules on natural amyloid template (i.e. plaque), is known to be critical for amyloid formation in vivo. To date, however, the use of 'natural' amyloid template to study amyloid deposition is extremely difficult and cumbersome. There is a critical need for high-throughput analytical system that enables a 'multi-factorial' investigation of a number of different environmental factors on amyloid deposition. The overall objective of this R21 proposal is to develop novel in vitro characterization methods that will lead to better understanding of amyloid deposition and to more effective screening of therapeutics. To meet the objective, four specific aims are proposed: (1) Design 'synthetic' templates suitable for amyloid deposition study in vitro, (2) Develop a microfluidic system for high-throughput analysis of amyloid deposition, (3) Validate amyloid deposition results obtained with microfluidic system, and (4) Conduct multi- factorial studies for screening inhibitors/accelerators of amyloid deposition/dissociation. The microfluidic system is intended to provide a novel way to study simultaneously how multiple environmental factors affect amyloid deposition/dissociation process while requiring only nano-liter amounts of amyloidogenic proteins and reagents. Results obtained from this research are expected to give deeper insight into the mechanism of amyloid deposition, as well as, to provide a powerful high-throughput protocol for screening therapeutics for amyloid-related diseases. ? ? ?