Abstract
Mutations in MAPT (microtubule-associated protein tau) cause frontotemporal dementia (FTD). MAPT mutations are associated with abnormal tau phosphorylation levels and accumulation of misfolded tau protein that can propagate between neurons ultimately leading to cell death (tauopathy). Recently, a p.A152T tau variant was identified as a risk factor for FTD, Alzheimer's disease, and synucleinopathies. Here we used induced pluripotent stem cells (iPSC) from a patient carrying this p.A152T variant to create a robust, functional cellular assay system for probing pathophysiological tau accumulation and phosphorylation. Using stably transduced iPSC-derived neural progenitor cells engineered to enable inducible expression of the pro-neural transcription factor Neurogenin 2 (Ngn2), we generated disease-relevant, cortical-like glutamatergic neurons in a scalable, high-throughput screening compatible format. Utilizing automated confocal microscopy, and an advanced image-processing pipeline optimized for analysis of morphologically complex human neuronal cultures, we report quantitative, subcellular localization-specific effects of multiple kinase inhibitors on tau, including ones under clinical investigation not previously reported to affect tau phosphorylation. These results demonstrate the potential for using patient iPSC-derived ex vivo models of tauopathy as genetically accurate, disease-relevant systems to probe tau biochemistry and support the discovery of novel therapeutics for tauopathies.
Highlights
Mutations in MAPT cause frontotemporal dementia (FTD)
A rare variant of tau, p.A152T, located in the proline-rich domain upstream of the microtubule-binding domain, has been identified as a risk factor for FTD, Alzheimer’s disease (AD) and synucleinopathies[13,14,15,16,17,18,19,20,21,22,23,24,25,26]. This variant of tau has decreased affinity for binding microtubules in vitro[13] and we have previously shown that induced pluripotent stem cells-derived neurons carrying this A152T variant (Tau-A152T) harbour a larger and more insoluble tau load than control n eurons[27]
We expand our work with induced pluripotent stem cells (iPSC) from an FTD subject diagnosed with progressive supranuclear palsy carrying this A152T variant as a genetically accurate cell model of tauopathy, in order to enable screening for novel therapeutics
Summary
Mutations in MAPT (microtubule-associated protein tau) cause frontotemporal dementia (FTD). An advanced imageprocessing pipeline optimized for analysis of morphologically complex human neuronal cultures, we report quantitative, subcellular localization-specific effects of multiple kinase inhibitors on tau, including ones under clinical investigation not previously reported to affect tau phosphorylation These results demonstrate the potential for using patient iPSC-derived ex vivo models of tauopathy as genetically accurate, disease-relevant systems to probe tau biochemistry and support the discovery of novel therapeutics for tauopathies. We expand our work with iPSC from an FTD subject diagnosed with progressive supranuclear palsy carrying this A152T variant as a genetically accurate cell model of tauopathy, in order to enable screening for novel therapeutics Using this model, we developed a rapid, reproducible, and scalable microwell neuronal differentiation system which consists of excitatory neurons generated by inducible expression of the pro-neural transcription factor Neurogenin 2 (Ngn[2]). This strategy will aid in expediting translational research in elucidating novel targets of therapeutic intervention for FTD-tau and other tauopathies[3]
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