A cellular model of amyloid precursor protein processing and amyloid-? peptide production.

ABSTRACT: A hallmark pathologic feature of Alzheimer's disease (AD) is accumulation of neuritic senile plaques in the brain parenchyma. Neurotoxic plaque cores are composed predominantly of amyloid-? (A?) peptides of 40 and 42 amino acids in length, formed by sequential cleavage of amyloid precursor protein (APP) by ?-, and ?-secretases. There is a great interest in approaches to modulate A? peptide production and develop therapeutic interventions to reduce A? levels to halt or slow the progression of neurodegeneration.We characterized and present the BE(2)-M17 human neuroblastoma cell line as a novel in vitro model of the APP-cleavage cascade to support future (1) functional studies of molecular regulators in A? production, and (2) high-throughput screening assays of new pharmacotherapeutics.In BE(2)-M17 cells, both RNA (i.e., RT-PCR, RNA sequencing) and protein analyses (i.e., Western blots, ELISA), show endogenous expression of critical components of the amyloidogenic pathway, APP-cleavage intermediates CTF83 and CTF99, and final cleavage products A?40 and A?42. We further report effects of retinoic acid-mediated differentiation on morphology and gene expression in this cell line.In contrast to primary isolates or other cell lines reported in current literature, BE(2)-M17 not only sustains baseline expression of the full contingent of APP-processing components, but also remains stably adherent during culture, facilitating experimental manipulations.Our evidence supports the use of BE(2)-M17 as a novel, human, cell-based model of the APP processing pathway that offers a potential streamlined approach to dissect molecular functions of endogenous regulatory pathways, and perform mechanistic studies to identify modulators of A? production.

SUBMITTER: Macias MP 

PROVIDER: S-EPMC3931259 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature