title: Probing sporadic and familial Alzheimer's disease using induced pluripotent stem cells.
abstract: Our understanding of Alzheimer's disease pathogenesis is currently limited by difficulties in obtaining live neurons from patients and the inability to model the sporadic form of the disease. It may be possible to overcome these challenges by reprogramming primary cells from patients into induced pluripotent stem cells (iPSCs). Here we reprogrammed primary fibroblasts from two patients with familial Alzheimer's disease, both caused by a duplication of the amyloid-β precursor protein gene (APP; termed APP(Dp)), two with sporadic Alzheimer's disease (termed sAD1, sAD2) and two non-demented control individuals into iPSC lines. Neurons from differentiated cultures were purified with fluorescence-activated cell sorting and characterized. Purified cultures contained more than 90% neurons, clustered with fetal brain messenger RNA samples by microarray criteria, and could form functional synaptic contacts. Virtually all cells exhibited normal electrophysiological activity. Relative to controls, iPSC-derived, purified neurons from the two APP(Dp) patients and patient sAD2 exhibited significantly higher levels of the pathological markers amyloid-β(1-40), phospho-tau(Thr 231) and active glycogen synthase kinase-3β (aGSK-3β). Neurons from APP(Dp) and sAD2 patients also accumulated large RAB5-positive early endosomes compared to controls. Treatment of purified neurons with β-secretase inhibitors, but not γ-secretase inhibitors, caused significant reductions in phospho-Tau(Thr 231) and aGSK-3β levels. These results suggest a direct relationship between APP proteolytic processing, but not amyloid-β, in GSK-3β activation and tau phosphorylation in human neurons. Additionally, we observed that neurons with the genome of one sAD patient exhibited the phenotypes seen in familial Alzheimer's disease samples. More generally, we demonstrate that iPSC technology can be used to observe phenotypes relevant to Alzheimer's disease, even though it can take decades for overt disease to manifest in patients.
authors.affiliation: Howard Hughes Medical Institute and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Mason A
authors.initials: MA
authors.lastname: Israel
authors.forename: Shauna H
authors.initials: SH
authors.lastname: Yuan
authors.forename: Cedric
authors.initials: C
authors.lastname: Bardy
authors.forename: Sol M
authors.initials: SM
authors.lastname: Reyna
authors.forename: Yangling
authors.initials: Y
authors.lastname: Mu
authors.forename: Cheryl
authors.initials: C
authors.lastname: Herrera
authors.forename: Michael P
authors.initials: MP
authors.lastname: Hefferan
authors.forename: Sebastiaan
authors.initials: S
authors.lastname: Van Gorp
authors.forename: Kristopher L
authors.initials: KL
authors.lastname: Nazor
authors.forename: Francesca S
authors.initials: FS
authors.lastname: Boscolo
authors.forename: Christian T
authors.initials: CT
authors.lastname: Carson
authors.forename: Louise C
authors.initials: LC
authors.lastname: Laurent
authors.forename: Martin
authors.initials: M
authors.lastname: Marsala
authors.forename: Fred H
authors.initials: FH
authors.lastname: Gage
authors.forename: Anne M
authors.initials: AM
authors.lastname: Remes
authors.forename: Edward H
authors.initials: EH
authors.lastname: Koo
authors.forename: Lawrence S B
authors.initials: LS
authors.lastname: Goldstein
mesh_terms: D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000378:metabolism* / Q000473:pathology*; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors / Q000378:metabolism; D016229:Amyloid beta-Peptides / Q000378:metabolism; D016564:Amyloid beta-Protein Precursor / Q000235:genetics / Q000378:metabolism; D001253:Astrocytes / Q000166:cytology; D015415:Biomarkers / Q000378:metabolism; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D018920:Coculture Techniques; D011992:Endosomes / Q000378:metabolism; D004789:Enzyme Activation; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism* / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D010446:Peptide Fragments / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011480:Protease Inhibitors / Q000494:pharmacology; D059748:Proteolysis; D016704:Synapsins / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism
chemical_list: D016229:Amyloid beta-Peptides; D016564:Amyloid beta-Protein Precursor; D015415:Biomarkers; D010446:Peptide Fragments; D010750:Phosphoproteins; D011480:Protease Inhibitors; D016704:Synapsins; C078592:amyloid beta-protein (1-40); D016875:tau Proteins; D038362:Glycogen Synthase Kinase 3; D053829:Amyloid Precursor Protein Secretases