科研之友
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摘要
Background: 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-beta (A beta) peptides of 40 and 42 amino acids in length, formed by sequential cleavage of amyloid precursor protein (APP) by beta-, and gamma-secretases. There is a great interest in approaches to modulate A beta peptide production and develop therapeutic interventions to reduce A beta levels to halt or slow the progression of neurodegeneration. New method: 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 beta production, and (2) high-throughput screening assays of new pharmacotherapeutics. Results: 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 beta 40 and A beta 42. We further report effects of retinoic acid-mediated differentiation on morphology and gene expression in this cell line. Comparison with existing method(s): 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. Conclusions: 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 beta production.
- 出版日期2014-2-15
