Abstract
SummaryOne potential therapeutic strategy for Alzheimer’s disease (AD) is to use antibodies that bind to small soluble protein aggregates to reduce their toxic effects. However, these therapies are rarely tested in human CSF before clinical trials because of the lack of sensitive methods that enable the measurement of aggregate-induced toxicity at low concentrations. We have developed highly sensitive single vesicle and single-cell-based assays that detect the Ca2+ influx caused by the CSF of individuals affected with AD and healthy controls, and we have found comparable effects for both types of samples. We also show that an extracellular chaperone clusterin; a nanobody specific to the amyloid-β peptide (Aβ); and bapineuzumab, a humanized monoclonal antibody raised against Aβ, could all reduce the Ca2+ influx caused by synthetic Aβ oligomers but are less effective in CSF. These assays could be used to characterize potential therapeutic agents in CSF before clinical trials.
Highlights
Protein misfolding and aggregation underlies a range of neurodegenerative diseases, including Alzheimer’s disease (AD)
(A) Quantitative ultrasensitive single vesicle assay to assess the ability of species (e.g., Ab42 aggregates or the species present in human cerebrospinal fluid (CSF)) to permeate the vesicle membrane
We have shown that even low levels of Ab42 oligomers can induce Ca2+ influx into neuronal cells by local dosing with a nanopipette (Figure 1B) (Drews et al, 2016)
Summary
Protein misfolding and aggregation underlies a range of neurodegenerative diseases, including Alzheimer’s disease (AD). We have in the past lacked the techniques needed to measure and quantify effectively such species present in vivo, for example, in human cerebrospinal fluid (CSF), where the concentrations are approximately 1 pM (Savage et al, 2014; Yang et al, 2015). This is an important problem since a number of therapeutic strategies are based on the use of antibodies or nanobodies to reduce the number of or the toxic effects of protein aggregates
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