Development of a Small D-Enantiomeric Alzheimer’s Amyloid-β Binding Peptide Ligand for Future In Vivo Imaging Applications

Funke, Susanne Aileen; Bartnik, Dirk; Glück, Julian M.; Piorkowska, Kasia; Wiesehan, Katja; Weber, Urs; Gulyás, Balázs; Halldin, Christer; Pfeifer, Andrea; Spenger, Christian; Muhs, Andreas; Willbold, Dieter

doi:10.1371/journal.pone.0041457

Cited by 23 publications

(19 citation statements)

References 31 publications

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“…It is crucial to highlight that this value is only an overall estimate of binding affinity across the entire spectrum of oligomers present in the oA␤42 preparation, which is a heterogeneous mixture of aggregates. The best fit occurred for a 1:1 binding model with drifting baseline correction ( 2 ϭ 0.214) and yielded a K D value of low micromolar value (11 M), indicating a moderate affinity between the 15M S.A. peptide and oA␤42, and similar to other reports of A␤-peptide interactions (34).…”

Section: M Sa Binds Pre-formed A␤42 Oligomers As Shown By Surfacsupporting

confidence: 85%

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

Barr

Verdile

Wijaya

et al. 2016

Journal of Biological Chemistry

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Although the formation of ␤-amyloid (A␤) deposits in the brain is a hallmark of Alzheimer disease (AD), the soluble oligomers rather than the mature amyloid fibrils most likely contribute to A␤ toxicity and neurodegeneration. Thus, the discovery of agents targeting soluble A␤ 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 A␤ 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 A␤42 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 A␤42, as shown by coimmunoprecipitation and surface plasmon resonance/Biacore analysis, with an affinity in the low micromolar range. Furthermore, this peptide bound fibrillar A␤42 and also stained plaques ex vivo in brain tissue from AD model mice. Given its multifaceted ability to target monomeric and aggregated A␤42 species, this candidate holds promise for novel preclinical AD imaging and therapeutic strategies. Alzheimer disease (AD)4 is a progressive neurodegenerative disorder, accounting for 50 -70% of late-onset dementia cases (2). The major biochemical hallmarks of AD are extracellular amyloid plaques and intracellular neurofibrillary tangles. The predominant species present in amyloid plaques is ␤-amyloid (A␤), a 4.5-kDa peptide generated through amyloidogenic processing (sequential cleavage by ␤-and ␥-secretases) of the parent amyloid precursor protein (APP). This produces two forms of A␤, comprising either 40 or 42 amino acids, where the relative amount of the longer 42-amino acid form (A␤42) is especially critical for AD progression, given its higher propensity to aggregate and form neurotoxic species (3, 4).In AD brain, the monomeric A␤42 peptide is known to aggregate and form various ordered assemblies, which precede plaque formation. These include low-n oligomers (dimers to octamers, reviewed in Ref. 5), high molecular weight oligomers such as A␤-derived diffusible ligands (6) and globulomers (7), protofibrils (8), and fibrils (9). Much evidence has indicated that soluble A␤42 oligomers, rather than mature amyloid plaques, correlate with disease severity (10, 11) and contribute to synapti...

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Section: M Sa Binds Pre-formed A␤42 Oligomers As Shown By Surfacsupporting

confidence: 85%

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

Barr

Verdile

Wijaya

et al. 2016

Journal of Biological Chemistry

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“…In this study we infused the hippocampus of Tg AD model mice for eight weeks with the small D-enantiomeric peptide D3 conjugated to FITC (D3- FITC) that has a high affinity for oligomeric Aβ 42 [23, 24, 26], and examined cognitive performance after 6.5 weeks, and the Aβ load after the end of the cognitive testing.…”

Section: Discussionmentioning

confidence: 99%

“…The identification of D3 is described elsewhere [24–27]. We chose to use D3-FITC (i.e., D3 conjugated with a FITC moiety via an additional C-terminal lysine; D3 sequence: rprtrlhthrnr) because we wanted to follow its fate in the animal and the FITC-label does, if at all, positively affect the D3 Aβ binding properties [30].…”

Section: Methodsmentioning

confidence: 99%

Treatment with D3 Removes Amyloid Deposits, Reduces Inflammation, and Improves Cognition in Aged AβPP/PS1 Double Transgenic Mice

Groen

Kadish

Funke

et al. 2013

JAD

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One of the characteristic pathological hallmarks of Alzheimer’s disease (AD) is neuritic plaques. The sequence of events leading to deposition of amyloid-β (Aβ) peptides in plaques is not clear. Here we investigate the effects of D3, an Aβ oligomer directed D-enantiomeric peptide that was obtained from a mirror image phage display selection against monomeric or small oligomeric forms of Aβ42, on Aβ deposition in aged AβPP/PS1 double transgenic AD-model mice. Using Alzet minipumps, we infused the brains of these AD model mice for 8 weeks with FITC-labeled D3, and examined the subsequent changes in pathology and cognitive deficits. Initial cognitive deficits are similar comparing control and D3-FITC-treated mice, but the treated mice show a significant improvement on the last day of testing. Further, we show that there is a substantial reduction in the amount of amyloid deposits in the animals treated with D3-FITC, compared to the control mice. Finally, the amount of activated microglia and astrocytes surrounding Aβ deposits is dramatically reduced in the D3-FITC-treated mice. Our findings demonstrate that treatments with the high affinity Aβ42 oligomer binding D-enantiomeric peptide D3 significantly decrease Aβ deposits and the associated inflammatory response, and improve cognition even when applied only at late stages and high age. Together, this suggests that the treatment reduces the level of Aβ peptide in the brains of AβPP/PS1 mice, possibly by increasing Aβ outflow from the brain. In conclusion, treatments with this D-peptide have great potential to be successful in AD patients.

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“…D1 peptide has the capacity to inhibit the Aβ peptide aggregation and to promote Aβ aggregates disaggregation. D1 is formed by D amino acids that are stable against endogenous proteases [12][13][14]. In the present research, we demonstrated that GNRs functionalized with the peptide D1 containing a spacer of polyethyleneglycol (PEG) (GNR-PEG-D1) promotes the enhancement of the fluorescence signal of CRANAD-2 associated with Aβ aggregates (Figure 3).…”

Section: Introductionmentioning

confidence: 58%

Section: Introductionmentioning

confidence: 99%

Gold Nanoparticles Mediate Improved Detection of β-amyloid Aggregates by Fluorescence

et al. 2020

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The early detection of the amyloid beta peptide aggregates involved in Alzheimer’s disease is crucial to test new potential treatments. In this research, we improved the detection of amyloid beta peptide aggregates in vitro and ex vivo by fluorescence combining the use of CRANAD-2 and gold nanorods (GNRs) by the surface enhancement fluorescence effect. We synthetized GNRs and modified their surface with HS-PEG-OMe and HS-PEG-COOH and functionalized them with the D1 peptide, which has the capability to selectively bind to amyloid beta peptide. For an in vitro detection of amyloid beta peptide, we co-incubated amyloid beta peptide aggregates with the probe CRANAD-2 and GNR-PEG-D1 observing an increase in the intensity of the fluorescence signal attributed to surface enhancement fluorescence. Furthermore, the surface enhancement fluorescence effect was observed in brain slices of transgenic mice with Alzheimer´s disease co-incubated with CRANAD-2 and GNR-PEG-D1. An increase in the fluorescence signal was observed allowing the detection of aggregates that cannot be detected with the single use of CRANAD-2. Gold nanoparticles allowed an improvement in the detection of the amyloid aggregated by fluorescence in vitro and ex vivo.

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Development of a Small D-Enantiomeric Alzheimer’s Amyloid-β Binding Peptide Ligand for Future In Vivo Imaging Applications

Cited by 23 publications

References 31 publications

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

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

Treatment with D3 Removes Amyloid Deposits, Reduces Inflammation, and Improves Cognition in Aged AβPP/PS1 Double Transgenic Mice

Gold Nanoparticles Mediate Improved Detection of β-amyloid Aggregates by Fluorescence

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