Dataset Information

Anti-A? drug screening platform using human iPS cell-derived neurons for the treatment of Alzheimer's disease.

ABSTRACT:

Background

Alzheimer's disease (AD) is a neurodegenerative disorder that causes progressive memory and cognitive decline during middle to late adult life. The AD brain is characterized by deposition of amyloid ? peptide (A?), which is produced from amyloid precursor protein by ?- and ?-secretase (presenilin complex)-mediated sequential cleavage. Induced pluripotent stem (iPS) cells potentially provide an opportunity to generate a human cell-based model of AD that would be crucial for drug discovery as well as for investigating mechanisms of the disease.

Methodology/principal findings

We differentiated human iPS (hiPS) cells into neuronal cells expressing the forebrain marker, Foxg1, and the neocortical markers, Cux1, Satb2, Ctip2, and Tbr1. The iPS cell-derived neuronal cells also expressed amyloid precursor protein, ?-secretase, and ?-secretase components, and were capable of secreting A? into the conditioned media. A? production was inhibited by ?-secretase inhibitor, ?-secretase inhibitor (GSI), and an NSAID; however, there were different susceptibilities to all three drugs between early and late differentiation stages. At the early differentiation stage, GSI treatment caused a fast increase at lower dose (A? surge) and drastic decline of A? production.

Conclusions/significance

These results indicate that the hiPS cell-derived neuronal cells express functional ?- and ?-secretases involved in A? production; however, anti-A? drug screening using these hiPS cell-derived neuronal cells requires sufficient neuronal differentiation.

SUBMITTER: Yahata N

PROVIDER: S-EPMC3184175 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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Publications

Anti-Aβ drug screening platform using human iPS cell-derived neurons for the treatment of Alzheimer's disease.

Yahata Naoki N Asai Masashi M Kitaoka Shiho S Takahashi Kazutoshi K Asaka Isao I Hioki Hiroyuki H Kaneko Takeshi T Maruyama Kei K Saido Takaomi C TC Nakahata Tatsutoshi T Asada Takashi T Yamanaka Shinya S Iwata Nobuhisa N Inoue Haruhisa H

PloS one 20110930 9

<h4>Background</h4>Alzheimer's disease (AD) is a neurodegenerative disorder that causes progressive memory and cognitive decline during middle to late adult life. The AD brain is characterized by deposition of amyloid β peptide (Aβ), which is produced from amyloid precursor protein by β- and γ-secretase (presenilin complex)-mediated sequential cleavage. Induced pluripotent stem (iPS) cells potentially provide an opportunity to generate a human cell-based model of AD that would be crucial for dru ...[more]

PMID: 21984949

Similar Datasets

Project description:Human induced pluripotent stem cell (iPSC)-derived neurons have been proposed to be a highly valuable cellular model for studying the pathomechanisms of Alzheimer's disease (AD). Studies employing patient-specific human iPSCs as models of familial and sporadic forms of AD described elevated levels of AD-related amyloid-? (A?). However, none of the present AD iPSC studies could recapitulate the synaptotoxic actions of A?, which are crucial early events in a cascade that eventually leads to vast brain degeneration. Here we established highly reproducible, human iPSC-derived cortical cultures as a cellular model to study the synaptotoxic effects of A?. We developed a highly efficient immunopurification procedure yielding immature neurons that express markers of deep layer cortical pyramidal neurons and GABAergic interneurons. Upon long-term cultivation, purified cells differentiated into mature neurons exhibiting the generation of action potentials and excitatory glutamatergic and inhibitory GABAergic synapses. Most interestingly, these iPSC-derived human neurons were strongly susceptible to the synaptotoxic actions of A?. Application of A? for 8 days led to a reduction in the overall FM4-64 and vGlut1 staining of vesicles in neurites, indicating a loss of vesicle clusters. A selective analysis of presynaptic vesicle clusters on dendrites did not reveal a significant change, thus suggesting that A? impaired axonal vesicle clusters. In addition, electrophysiological patch-clamp recordings of AMPA receptor-mediated miniature EPSCs revealed an A?-induced reduction in amplitudes, indicating an impairment of postsynaptic AMPA receptors. A loss of postsynaptic AMPA receptor clusters was confirmed by immunocytochemical stainings for GluA1. Incubation with A? for 8 days did not result in a significant loss of neurites or cell death. In summary, we describe a highly reproducible cellular AD model based on human iPSC-derived cortical neurons that enables the mechanistic analysis of A?-induced synaptic pathomechanisms and the development of novel therapeutic approaches.