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FRET-based calcium imaging: a tool for high-throughput/content phenotypic drug screening in Alzheimer disease.

Authors :
Honarnejad K
Kirsch AK
Daschner A
Szybinska A
Kuznicki J
Herms J
Source :
Journal of biomolecular screening [J Biomol Screen] 2013 Dec; Vol. 18 (10), pp. 1309-20.
Publication Year :
2013

Abstract

Perturbed intracellular store calcium homeostasis is suggested to play a major role in the pathophysiology of Alzheimer disease (AD). A number of mechanisms have been suggested to underlie the impairment of endoplasmic reticulum calcium homeostasis associated with familial AD-linked presenilin 1 mutations (FAD-PS1). Without aiming at specifically targeting any of those pathophysiological mechanisms in particular, we rather performed a high-throughput phenotypic screen to identify compounds that can reverse the exaggerated agonist-evoked endoplasmic reticulum calcium release phenotype in HEK293 cells expressing FAD-PS1. For that purpose, we developed a fully automated high-throughput calcium imaging assay using a fluorescence resonance energy transfer-based calcium indicator at single-cell resolution. This novel robust assay offers a number of advantages compared with the conventional calcium measurement screening technologies. The assay was employed in a large-scale screen with a library of diverse compounds comprising 20,000 low-molecular-weight molecules, which resulted in the identification of 52 primary hits and 4 lead structures. In a secondary assay, several hits were found to alter the amyloid β (Aβ) production. In view of the recent failure of AD drug candidates identified by target-based approaches, such a phenotypic drug discovery paradigm may present an attractive alternative for the identification of novel AD therapeutics.

Details

Language :
English
ISSN :
1552-454X
Volume :
18
Issue :
10
Database :
MEDLINE
Journal :
Journal of biomolecular screening
Publication Type :
Academic Journal
Accession number :
24221842
Full Text :
https://doi.org/10.1177/1087057113502672