Validation and Characterization of a Novel Peptide That Binds Monomeric and Aggregated β-Amyloid and Inhibits the Formation of Neurotoxic Oligomers

J Biol Chem. 2016 Jan 8;291(2):547-59. doi: 10.1074/jbc.M115.679993. Epub 2015 Nov 4.

Abstract

Although the formation of β-amyloid (Aβ) deposits in the brain is a hallmark of Alzheimer disease (AD), the soluble oligomers rather than the mature amyloid fibrils most likely contribute to Aβ toxicity and neurodegeneration. Thus, the discovery of agents targeting soluble Aβ oligomers is highly desirable for early diagnosis prior to the manifestation of a clinical AD phenotype and also more effective therapies. We have previously reported that a novel 15-amino acid peptide (15-mer), isolated via phage display screening, targeted Aβ and attenuated its neurotoxicity (Taddei, K., Laws, S. M., Verdile, G., Munns, S., D'Costa, K., Harvey, A. R., Martins, I. J., Hill, F., Levy, E., Shaw, J. E., and Martins, R. N. (2010) Neurobiol. Aging 31, 203-214). The aim of the current study was to generate and biochemically characterize analogues of this peptide with improved stability and therapeutic potential. We demonstrated that a stable analogue of the 15-amino acid peptide (15M S.A.) retained the activity and potency of the parent peptide and demonstrated improved proteolytic resistance in vitro (stable to t = 300 min, c.f. t = 30 min for the parent peptide). This candidate reduced the formation of soluble Aβ42 oligomers, with the concurrent generation of non-toxic, insoluble aggregates measuring up to 25-30 nm diameter as determined by atomic force microscopy. The 15M S.A. candidate directly interacted with oligomeric Aβ42, as shown by coimmunoprecipitation and surface plasmon resonance/Biacore analysis, with an affinity in the low micromolar range. Furthermore, this peptide bound fibrillar Aβ42 and also stained plaques ex vivo in brain tissue from AD model mice. Given its multifaceted ability to target monomeric and aggregated Aβ42 species, this candidate holds promise for novel preclinical AD imaging and therapeutic strategies.

Keywords: Alzheimer disease; amyloid-β (AB); imaging; neurodegeneration; peptides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Intravenous
  • Alzheimer Disease / metabolism
  • Alzheimer Disease / pathology
  • Amyloid / metabolism*
  • Animals
  • Brain / metabolism
  • Brain / pathology
  • Disease Models, Animal
  • Humans
  • Immunohistochemistry
  • Immunoprecipitation
  • Male
  • Mice, Transgenic
  • Neurotoxins / toxicity*
  • Peptides / metabolism*
  • Protein Aggregates / drug effects*
  • Protein Multimerization / drug effects*
  • Protein Stability / drug effects
  • Reproducibility of Results
  • Solubility
  • Surface Plasmon Resonance
  • Tritium / metabolism

Substances

  • Amyloid
  • Neurotoxins
  • Peptides
  • Protein Aggregates
  • Tritium