Familial Alzheimer’s disease (fAD) mutations alter amyloid precursor protein (APP) cleavage by γ-secretase, increasing the proportion of longer amyloidogenic amyloid-β (Aβ) peptides. Using five control induced pluripotent stem cell (iPSC) lines and seven iPSC lines generated from fAD patients, we investigated the effects of mutations on the Aβ secretome in human neurons generated in 2D and 3D. We also analysed matched CSF, post-mortem brain tissue, and iPSCs from the same participant with the APP V717I mutation. All fAD mutation lines demonstrated an increased Aβ42:40 ratio relative to controls, yet displayed varied signatures for Aβ43, Aβ38, and short Aβ fragments. We propose four qualitatively distinct mechanisms behind raised Aβ42:40. (1) APP V717I mutations alter γ-secretase cleavage site preference. Whereas, distinct presenilin 1 (PSEN1) mutations lead to either (2) reduced γ-secretase activity, (3) altered protein stability or (4) reduced PSEN1 maturation, all culminating in reduced γ-secretase carboxypeptidase-like activity. These data support Aβ mechanistic tenets in a human physiological model and substantiate iPSC-neurons for modelling fAD.
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Selkoe DJ, Hardy J. The amyloid hypothesis of Alzheimer’s disease at 25 years. EMBO Mol Med. 2016;8:595–608.
Van Cauwenberghe C, Van Broeckhoven C, Sleegers K. The genetic landscape of Alzheimer disease: clinical implications and perspectives. Genet Med. 2016;18:421–30.
O’Brien RJ, Wong PC. Amyloid precursor protein processing and Alzheimer’s disease. Annu Rev Neurosci. 2011;34:185–204.
Fernandez MA, Biette KM, Dolios G, Seth D, Wang R, Wolfe MS. Transmembrane substrate determinants for γ-secretase processing of APP CTFβ. Biochemistry. 2016;55:5675–88.
Takami M, Nagashima Y, Sano Y, Ishihara S, Morishima-Kawashima M, Funamoto S, et al. Gamma-Secretase: successive tripeptide and tetrapeptide release from the transmembrane domain of -carboxyl terminal fragment. J Neurosci. 2009;29:13042–52.
Matsumura N, Takami M, Okochi M, Wada-Kakuda S, Fujiwara H, Tagami S, et al. γ-Secretase associated with lipid rafts: multiple interactive pathways in the stepwise processing of β-carboxylterminal fragment. J Biol Chem. 2014;289:5109–21.
Fernandez MA, Klutkowski JA, Freret T, Wolfe MS. Alzheimer presenilin-1 mutations dramatically reduce trimming of long amyloid β-peptides (Aβ) by γ-secretase to increase 42-to-40-residue Aβ. J Biol Chem. 2014;289:31043–52.
Chavez-Gutierrez L, Bammens L, Benilova I, Vandersteen A, Benurwar M, Borgers M, et al. The mechanism of gamma-secretase dysfunction in familial Alzheimer disease. EMBO J. 2012;31:2261–74.
Szaruga M, Veugelen S, Benurwar M, Lismont S, Sepulveda-Falla D, Lleo A, et al. Qualitative changes in human γ-secretase underlie familial Alzheimer’s disease. J Exp Med. 2015;212:2003–13.
Slemmon JR, Shapiro A, Mercken M, Streffer J, Romano G, Andreasen N, et al. Impact of cerebrospinal fluid matrix on the detection of Alzheimer’s disease with Abeta42 and influence of disease on the total-Abeta42/Abeta40 ratio. J Neurochem. 2015;135:1049–58.
Vanderstichele H, Bibl M, Engelborghs S, Le Bastard N, Lewczuk P, Molinuevo JL, et al. Standardization of preanalytical aspects of cerebrospinal fluid biomarker testing for Alzheimer’s disease diagnosis: a consensus paper from the Alzheimer’s Biomarkers Standardization Initiative. Alzheimers Dement. 2012;8:65–73.
Toombs J, Paterson RW, Schott JM, Zetterberg H. Amyloid-beta 42 adsorption following serial tube transfer. Alzheimers Res Ther. 2014;6:5.
Janelidze S, Zetterberg H, Mattsson N, Palmqvist S, Vanderstichele H, Lindberg O, et al. CSF Aβ42/Aβ40 and Aβ42/Aβ38 ratios: better diagnostic markers of Alzheimer disease. Ann Clin Transl Neurol. 2016;3:154–65.
Dorey A, Perret-Liaudet A, Tholance Y, Fourier A, Quadrio I. Cerebrospinal fluid Aβ40 improves the interpretation of Aβ42 concentration for diagnosing Alzheimer’s disease. Front Neurol. 2015;6:247.
Blennow K, Zetterberg H, Fagan AM. Fluid biomarkers in Alzheimer disease. Cold Spring Harb Perspect Med. 2012;2:a006221.
Arber C, Lovejoy C, Wray S. Stem cell models of Alzheimer’s disease: progress and challenges. Alzheimers Res Ther. 2017;9:42.
Yagi T, Ito D, Okada Y, Akamatsu W, Nihei Y, Yoshizaki T, et al. Modeling familial Alzheimer’s disease with induced pluripotent stem cells. Hum Mol Genet. 2011;20:4530–9.
Mahairaki V, Ryu J, Peters A, Chang Q, Li T, Park TS, et al. Induced pluripotent stem cells from familial Alzheimer’s disease patients differentiate into mature neurons with amyloidogenic properties. Stem Cells Dev. 2014;23:2996–3010.
Sproul AA, Jacob S, Pre D, Kim SH, Nestor MW, Navarro-Sobrino M, et al. Characterization and molecular profiling of PSEN1 familial Alzheimer’s disease iPSC-Derived neural progenitors. PLoS ONE 2014;9:e84547.
Woodruff G, Young JE, Martinez FJ, Buen F, Gore A, Kinaga J, et al. The presenilin-1 ΔE9 mutation results in reduced γ-secretase activity, but not total loss of PS1 function, in isogenic human stem cells. Cell Rep. 2013;5:974–85.
Moore S, Evans LDB, Andersson T, Portelius E, Smith J, Dias TB, et al. APP metabolism regulates tau proteostasis in human cerebral cortex neurons. Cell Rep. 2015;11:689–96.
Ochalek A, Mihalik B, Avci HX, Chandrasekaran A, Téglási A, Bock I, et al. Neurons derived from sporadic Alzheimer’s disease iPSCs reveal elevated TAU hyperphosphorylation, increased amyloid levels, and GSK3B activation. Alzheimers Res Ther. 2017;9:90.
Sun L, Zhou R, Yang G, Shi Y. Analysis of 138 pathogenic mutations in presenilin-1 on the in vitro production of Aβ42 and Aβ40 peptides by γ-secretase. Proc Natl Acad Sci USA. 2017;114:E476–85.
Muratore CR, Rice HC, Srikanth P, Callahan DG, Shin T, Benjamin LNP, et al. The familial Alzheimer’s disease APPV717I mutation alters APP processing and Tau expression in iPSC-derived neurons. Hum Mol Genet. 2014;23:3523–36.
Israel MA, Yuan SH, Bardy C, Reyna SM, Mu Y, Herrera C, et al. Probing sporadic and familial Alzheimer’s disease using induced pluripotent stem cells. Nature. 2012;482:216–20.
Lancaster MA, Knoblich J. Generation of cerebral organoids from human pluripotent stem. Nat Protoc. 2014;10:2329–40.
Raja WK, Mungenast AE, Lin Y-T, Ko T, Abdurrob F, Seo J, et al. Self-organizing 3D human neural tissue derived from induced pluripotent stem cells recapitulate Alzheimer’s disease phenotypes. PLoS ONE 2016;11:e0161969.
Chen M, Lee HK, Moo L, Hanlon E, Stein T, Xia W. Common proteomic profiles of induced pluripotent stem cell-derived three-dimensional neurons and brain tissue from Alzheimer patients. J Proteom. 2018;182:21–33.
Wray S, Self M, Lewis PA, Taanman J-W, Ryan NS, Mahoney CJ, et al. Creation of an open-access, mutation-defined fibroblast resource for neurological disease research. PLoS ONE 2012;7:e43099.
Okita K, Matsumura Y, Sato Y, Okada A, Morizane A, Okamoto S, et al. A more efficient method to generate integration-free human iPS cells. Nat Methods. 2011;8:409–12.
Shi Y, Kirwan P, Livesey FJ. Directed differentiation of human pluripotent stem cells to cerebral cortex neurons and neural networks. Nat Protoc. 2012;7:1836–46.
Portelius E, Tran AJ, Andreasson U, Persson R, Brinkmann G, Zetterberg H, et al. Characterization of amyloid β peptides in cerebrospinal fluid by an automated immunoprecipitation procedure followed by mass spectrometry. J Proteome Res. 2007;6:4433–9.
Germain PL, Testa G. Taming human genetic variability: transcriptomic meta-analysis guides the experimental design and interpretation of iPSC-based disease modeling. Stem Cell Rep. 2017;8:1784–96.
British Standards Institution. Accuracy (trueness and precision) of measurement methods and results. Part 2: Basic method for the determination of repeatability and reproducibility of a standard measurement method [Internet]. Mol. Ecol. 1994 Available from: http://www.iso.org/iso/iso_catalogue/catalogue_tc/catalogue_detail.htm?csnumber=11833. Accessed 11 Feb. 2017.
Bergstrom P, Agholme L, Nazir FH, Satir TM, Toombs J, Wellington H, et al. Amyloid precursor protein expression and processing are differentially regulated during cortical neuron differentiation. Sci Rep. 2016;6:29200.
Veugelen S, Saito T, Saido TC, Chávez-Gutiérrez L, De Strooper B. Familial Alzheimer’s disease mutations in presenilin generate amyloidogenic Aβ peptide seeds. Neuron. 2016;90:410–6.
Saito T, Suemoto T, Brouwers N, Sleegers K, Funamoto S, Mihira N, et al. Potent amyloidogenicity and pathogenicity of Aβ43. Nat Neurosci. 2011;14:1023–32.
Szaruga M, Munteanu B, Lismont S, Veugelen S, Horré K, Mercken M, et al. Alzheimer’s-causing mutations shift Aβ length by destabilizing γ-secretase-Aβn interactions. Cell. 2017;170:443–.e14.
Yan R, Han P, Miao H, Greengard P, Xu H. The transmembrane domain of the Alzheimer’s β-secretase (BACE1) determines its late Golgi localization and access to β-amyloid precursor protein (APP) substrate. J Biol Chem. 2001;276:36788–96.
Shi X-P, Tugusheva K, Bruce JE, Lucka A, Wu G-X, Chen-Dodson E, et al. Beta-secretase cleavage at amino acid residue 34 in the amyloid beta peptide is dependent upon gamma-secretase activity. J Biol Chem. 2003;278:21286–94.
Portelius E, Price E, Brinkmalm G, Stiteler M, Olsson M, Persson R, et al. A novel pathway for amyloid precursor protein processing. Neurobiol Aging. 2011;32:1090–8.
Eckman EA, Reed DK, Eckman CB. Degradation of the Alzheimer’s amyloid beta peptide by endothelin- converting enzyme. J Biol Chem. 2001;276:24540–8.
Siegel G, Gerber H, Koch P, Bruestle O, Fraering PC, Rajendran L. The Alzheimer’s disease γ-secretase generates higher 42:40 ratios for β-amyloid than for p3 peptides. Cell Rep. 2017;19:1967–76.
Wanngren J, Lara P, Öjemalm K, Maioli S, Moradi N, Chen L, et al. Changed membrane integration and catalytic site conformation are two mechanisms behind the increased Aβ42/Aβ40 ratio by presenilin 1 familial Alzheimer-linked mutations. FEBS Open Bio. 2014;4:393–406.
Anand P, Nandel FS, Hansmann UHE. The Alzheimer beta-amyloid (Abeta(1-39)) dimer in an implicit solvent. J Chem Phys. 2008;129:195102.
Cloe AL, Orgel JPRO, Sachleben JR, et al. The Japanese mutant Aβ (ΔE22-Aβ 1-39) forms fibrils instantaneously, with low-thioflavin T fluorescence: seeding of wild-type Aβ 1-40 into atypical fibrils by Δe22- Aβ 1-39. Biochemistry. 2011;50:2026–39.
Takagi-Niidome S, Sasaki T, Osawa S, Sato T, Morishima K, Cai T, et al. Cooperative roles of hydrophilic loop 1 and the C-terminus of presenilin 1 in the substrate-gating mechanism of gamma-secretase. J Neurosci. 2015;35:2646–56.
Somavarapu AK, Kepp KP. The dynamic mechanism of presenilin-function: sensitive gate dynamics and loop unplugging control protein access. Neurobiol Dis. 2016;89:147–56.
Verdile G, Gnjec A, Miklossy J, Fonte J, Veurink G, Bates K, et al. Protein markers for Alzheimer in the frontal cortex and cerebellum. Neurology. 2004;63:1385–92.
Mathews PM, Cataldo AM, Kao BH, Rudnicki AG, Qin X, Yang JL, et al. Brain expression of presenilins in sporadic and early-onset, familial Alzheimer’s disease. Mol Med. 2000;6:878–91.
Moore BD, Martin J, de Mena L, Sanchez J, Cruz PE, Ceballos-Diaz C, et al. Short Aβ peptides attenuate Aβ42 toxicity in vivo. J Exp Med. 2018;215:283–301.
Ryan NS, Nicholas JM, Weston PSJ, Liang Y, Lashley T, Guerreiro R, et al. Clinical phenotype and genetic associations in autosomal dominant familial Alzheimer’s disease: a case series. Lancet Neurol. 2016;15:1326–35.
We gratefully acknowledge the support of the Leonard Wolfson Experimental Neurology Centre, the NIHR UCL Hospitals Biomedical Research Centre. The Dementia Research Centre is an Alzheimer’s Research UK Coordinating Centre. SW is supported by an Alzheimer’s Research UK Senior Research Fellowship (ARUK-SRF2016B-2). NF acknowledges the support of the UK Dementia Research Institute at UCL. NSR is supported by a University of London Chadburn Academic Clinical Lectureship in Medicine. The research leading to these results has received support from the Innovative Medicines Initiative Joint Undertaking under grant agreement no. 115439, resources of which are composed of financial contribution from the European Union’s Seventh Framework Programme (FP7/2007–2013) and EFPIA companies’ in kind contribution. This publication reflects only the author’s views and neither the IMI JU nor EFPIA nor the European Commission are liable for any use that may be made of the information contained therein. This work was supported by the UK Medical Research Council funding to the MRC Dementia Platform UK (MR/M02492X/1) and Medical Research Council core funding to the High-Content Biology Platform at the MRC-UCL LMCB university unit (MC_U12266B). We would like to thank Dr. Rita Louro Guerreiro, Lee Darwent, and Celia Kun Rodrigues for help with sequencing of iPSC clones.
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Gamma secretase modulators and BACE inhibitors reduce Aβ production without altering gene expression in Alzheimer's disease iPSC-derived neurons and mice
Molecular and Cellular Neuroscience (2019)
Nature Reviews Neurology (2019)