Development and Implementation of a High-Throughput Compound Screening Assay for Targeting Disrupted ER Calcium Homeostasis in Alzheimer%26apos;s Disease

作者：Honarnejad Kamran; Daschner Alexander; Giese Armin; Zall Andrea; Schmidt Boris; Szybinska Aleksandra; Kuznicki Jacek; Herms Jochen^*

来源：PLos One, 2013, 8(11): e80645.

DOI：10.1371/journal.pone.0080645

Disrupted intracellular calcium homeostasis is believed to occur early in the cascade of events leading to Alzheimer%26apos;s disease (AD) pathology. Particularly familial AD mutations linked to Presenilins result in exaggerated agonist-evoked calcium release from endoplasmic reticulum (ER). Here we report the development of a fully automated high-throughput calcium imaging assay utilizing a genetically-encoded FRET-based calcium indicator at single cell resolution for compound screening. The established high-throughput screening assay offers several advantages over conventional high-throughput calcium imaging technologies. We employed this assay for drug discovery in AD by screening compound libraries consisting of over 20,000 small molecules followed by structure-activity-relationship analysis. This led to the identification of Bepridil, a calcium channel antagonist drug in addition to four further lead structures capable of normalizing the potentiated FAD-PS1-induced calcium release from ER. Interestingly, it has recently been reported that Bepridil can reduce A beta production by lowering BACE1 activity. Indeed, we also detected lowered A beta, increased sAPP alpha and decreased sAPP beta fragment levels upon Bepridil treatment. The latter findings suggest that Bepridil may provide a multifactorial therapeutic modality for AD by simultaneously addressing multiple aspects of the disease.

• 出版日期2013-11-15