title: Genetic inhibition of phosphorylation of the translation initiation factor eIF2α does not block Aβ-dependent elevation of BACE1 and APP levels or reduce amyloid pathology in a mouse model of Alzheimer's disease.
abstract: β-site amyloid precursor protein (APP) cleaving enzyme 1 (BACE1) initiates the production of β-amyloid (Aβ), the major constituent of amyloid plaques in Alzheimer's disease (AD). BACE1 is elevated ∼2-3 fold in AD brain and is concentrated in dystrophic neurites near plaques, suggesting BACE1 elevation is Aβ-dependent. Previously, we showed that phosphorylation of the translation initiation factor eIF2α de-represses translation of BACE1 mRNA following stress such as energy deprivation. We hypothesized that stress induced by Aβ might increase BACE1 levels by the same translational mechanism involving eIF2α phosphorylation. To test this hypothesis, we used three different genetic strategies to determine the effects of reducing eIF2α phosphorylation on Aβ-dependent BACE1 elevation in vitro and in vivo: 1) a two-vector adeno-associated virus (AAV) system to express constitutively active GADD34, the regulatory subunit of PP1c eIF2α phosphatase; 2) a non-phosphorylatable eIF2α S51A knockin mutation; 3) a BACE1-YFP transgene lacking the BACE1 mRNA 5' untranslated region (UTR) required for eIF2α translational regulation. The first two strategies were used in primary neurons and 5XFAD transgenic mice, while the third strategy was employed only in 5XFAD mice. Despite very effective reduction of eIF2α phosphorylation in both primary neurons and 5XFAD brains, or elimination of eIF2α-mediated regulation of BACE1-YFP mRNA translation in 5XFAD brains, Aβ-dependent BACE1 elevation was not decreased. Additionally, robust inhibition of eIF2α phosphorylation did not block Aβ-dependent APP elevation in primary neurons, nor did it reduce amyloid pathology in 5XFAD mice. We conclude that amyloid-associated BACE1 elevation is not caused by translational de-repression via eIF2α phosphorylation, but instead appears to involve a post-translational mechanism. These definitive genetic results exclude a role for eIF2α phosphorylation in Aβ-dependent BACE1 and APP elevation. We suggest a vicious pathogenic cycle wherein Aβ42 toxicity induces peri-plaque BACE1 and APP accumulation in dystrophic neurites leading to exacerbated Aβ production and plaque progression.
authors.affiliation: Department of Cell and Molecular Biology, Northwestern University, Feinberg School of Medicine, Chicago, Illinois, United States of America.
authors.forename: Katherine R
authors.initials: KR
authors.lastname: Sadleir
authors.affiliation: Department of Neurology, Massachusetts General Hospital, Charlestown, Massachusetts, United States of America.
authors.forename: William A
authors.initials: WA
authors.lastname: Eimer
authors.affiliation: Sanford|Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: Randal J
authors.initials: RJ
authors.lastname: Kaufman
authors.affiliation: Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, United States of America.
authors.forename: Pavel
authors.initials: P
authors.lastname: Osten
authors.affiliation: Department of Cell and Molecular Biology, Northwestern University, Feinberg School of Medicine, Chicago, Illinois, United States of America.
authors.forename: Robert
authors.initials: R
authors.lastname: Vassar
mesh_terms: D020121:5' Untranslated Regions; D000544:Alzheimer Disease / Q000378:metabolism / Q000473:pathology*; D019943:Amino Acid Substitution; D053829:Amyloid Precursor Protein Secretases / Q000235:genetics / Q000378:metabolism*; D016229:Amyloid beta-Peptides / Q000378:metabolism*; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics / Q000378:metabolism*; D055879:Gene Knock-In Techniques; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010766:Phosphorylation; D054645:Protein Phosphatase 1 / Q000235:genetics / Q000378:metabolism
chemical_list: D020121:5' Untranslated Regions; D016229:Amyloid beta-Peptides; D016564:Amyloid beta-Protein Precursor; D015852:Eukaryotic Initiation Factor-2; C513256:Ppp1r15a protein, mouse; D054645:Protein Phosphatase 1; D053829:Amyloid Precursor Protein Secretases