Development of a high-throughput screen targeting caspase-8-mediated cleavage of the amyloid precursor protein

Matthew J. Hart, Marcie Glicksman, Min Liu, Mohit K. Sharma, Greg Cuny, Veronica Galvan

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Caspases, effectors of apoptosis, are key mediators of neuronal death in several neurodegenerative diseases. Caspase-8 and caspase-6 have been implicated in the pathogenesis of amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, and Alzheimer's disease (AD). β-Amyloid precursor protein (APP) is cleaved at Asp664 in its intracellular domain by caspase-8. We and other laboratories recently showed that obliteration of the caspase cleavage site on APP alleviates functional AD-like deficits in a mouse model. Therefore, caspase cleavage of APP constitutes a potential novel target for therapeutic intervention. To identify chemical inhibitors of caspase-8 cleavage, we screened a subset of the chemical library at the Harvard NeuroDiscovery Center's Laboratory for Drug Discovery in Neurodegeneration. We show that caspase-8, but not caspase-1, -3, or -9, cleaves a biotinylated peptide derived from APP at Asp664, and we report the development of a sensitive high-throughput assay for caspase-8 cleavage of APP and the use of that assay for the identification of specific small molecule "hit" compounds that potently inhibit Asp664 cleavage of APP. Furthermore, we demonstrate that one of these compounds (LDN-0021835) inhibits the cleavage of APP at Asp664 in cell-based assays.

Original languageEnglish (US)
Pages (from-to)467-476
Number of pages10
JournalAnalytical Biochemistry
Volume421
Issue number2
DOIs
StatePublished - Feb 15 2012

Keywords

  • Alzheimer's disease
  • Amyloid precursor protein
  • Caspase-8
  • High-throughput screening
  • Medicinal chemistry
  • Neurodegeneration

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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