Development of a High Throughput Drug Screening Assay for the Detection of Changes in Tau Levels - Proof of Concept with HSP90 inhibitors

Chad   A.   Dickey; Jason      Eriksen; Adeela      Kamal; Francis      Burrows; Srinivas      Kasibhatla; Christopher   B.   Eckman; Mike      Hutton; Leonard      Petrucelli

Abstract

Therapeutic development for Alzheimers disease has largely focused on the removal of beta amyloid because of its suggested role in the primary agent in initiating the disease process. However, with the recent discovery of mutations that result as pathologic buildup of tau in the absence of amyloid pathology, tau is beginning to be recognized as a potential target for drug discovery. We have developed a high-throughput drug screening method that allows for direct intracellular quantitation of tau protein species, enabling the fast, reliable detection of these changes. We have identified a family of small, blood brain barrier penetrant heat shock protein 90 inhibitors that significantly reduce tau protein levels in vitro. Western blot analysis demonstrated a clear inverse correlation between the tau levels and the increase in HSP27, HSP40 and HSP90. Modifications to this assay will further allow the specific analysis of pathologically relevant species. Using this assay, we have demonstrated that a class of HSP90 inhibitors is able to significantly lower intracellular tau levels most likely through induction of a heat shock response.

Keywords: alzheimer, ’, s disease (ad), neurodegeneration, neurofibrillary tangles (nft), nmda receptor antagonist, amyloid precursor protein, tau knockout mice, expression