Mimi P Macias1, Amanda M Gonzales2, Ashley L Siniard3, Aaron W Walker4, Jason J Corneveaux5, Matthew J Huentelman6, Marwan N Sabbagh7, Boris Decourt8. 1. Haldeman Laboratory of Molecular Diagnostics and Therapeutics, Banner Sun Health Research Institute, 10515 W, Santa Fe Drive, Sun City, AZ 85351, USA. Electronic address: mimi.macias@bannerhealth.com. 2. Haldeman Laboratory of Molecular Diagnostics and Therapeutics, Banner Sun Health Research Institute, 10515 W, Santa Fe Drive, Sun City, AZ 85351, USA. Electronic address: amanda.gonzales@bannerhealth.com. 3. Neurogenomics Division, The Translational Genomics Research Institute, 445N, Fifth Street, Phoenix, AZ 85004, USA. Electronic address: asiniard@tgen.org. 4. Haldeman Laboratory of Molecular Diagnostics and Therapeutics, Banner Sun Health Research Institute, 10515 W, Santa Fe Drive, Sun City, AZ 85351, USA. Electronic address: aaron.walker@bannerhealth.com. 5. Neurogenomics Division, The Translational Genomics Research Institute, 445N, Fifth Street, Phoenix, AZ 85004, USA. Electronic address: jcorneveaux@tgen.org. 6. Neurogenomics Division, The Translational Genomics Research Institute, 445N, Fifth Street, Phoenix, AZ 85004, USA. Electronic address: mhuentelman@tgen.org. 7. Haldeman Laboratory of Molecular Diagnostics and Therapeutics, Banner Sun Health Research Institute, 10515 W, Santa Fe Drive, Sun City, AZ 85351, USA. Electronic address: marwan.sabbagh@bannerhealth.com. 8. Haldeman Laboratory of Molecular Diagnostics and Therapeutics, Banner Sun Health Research Institute, 10515 W, Santa Fe Drive, Sun City, AZ 85351, USA. Electronic address: boris.decourt@bannerhealth.com.
Abstract
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-β (Aβ) peptides of 40 and 42 amino acids in length, formed by sequential cleavage of amyloid precursor protein (APP) by β-, and γ-secretases. There is a great interest in approaches to modulate Aβ peptide production and develop therapeutic interventions to reduce Aβ 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 Aβ 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 Aβ40 and Aβ42. 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 Aβ production.
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-β (Aβ) peptides of 40 and 42 amino acids in length, formed by sequential cleavage of amyloid precursor protein (APP) by β-, and γ-secretases. There is a great interest in approaches to modulate Aβ peptide production and develop therapeutic interventions to reduce Aβ levels to halt or slow the progression of neurodegeneration. NEW METHOD: We characterized and present the BE(2)-M17 humanneuroblastoma cell line as a novel in vitro model of the APP-cleavage cascade to support future (1) functional studies of molecular regulators in Aβ 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 Aβ40 and Aβ42. 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 Aβ production.
Authors: Boris Decourt; Amanda Gonzales; Thomas G Beach; Michael Malek-Ahmadi; Aaron Walker; Lucia Sue; Douglas G Walker; Marwan N Sabbagh Journal: Curr Alzheimer Res Date: 2013-03 Impact factor: 3.498
Authors: G K Gouras; H Xu; J N Jovanovic; J D Buxbaum; R Wang; P Greengard; N R Relkin; S Gandy Journal: J Neurochem Date: 1998-11 Impact factor: 5.372
Authors: Matt Huentelman; Matthew De Both; Wayne Jepsen; Ignazio S Piras; Joshua S Talboom; Mari Willeman; Eric M Reiman; John Hardy; Amanda J Myers Journal: Sci Rep Date: 2019-07-03 Impact factor: 4.379