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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 gene1 (APP; termed APPDp), 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 APPDp 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 APPDp 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.

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Change history

  • 08 February 2012

    An addition was made to Acknowlegements.

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Primary accessions

Gene Expression Omnibus

Data deposits

Data have been deposited in the Gene ExpressionOmnibus under accession GSE34879.

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Acknowledgements

We thank D. Galasko, M. Sundsmo, J. Rivera, J. Fontaine, C. Gigliotti and B. Yu at the University of California, San Diego (UCSD) Alzheimer’s Disease Research Center for patient samples and data (grant AGO 5131); S. Dowdy and N. Yoshioka for viral vectors; B. Balderas at BD Biosciences for antibodies; C. Santucci and S. Nguyen for teratoma assay assistance; the UCSD Neuroscience Microscopy Shared Facility (grant P30 NS047101); and Planned Parenthood of the Pacific Southwest for fetal brain specimens. Funding was from California Institute of Regenerative Medicine (CIRM) comprehensive grants (M.M., F.H.G., L.S.B.G.), CIRM predoctoral fellowship (M.A.I.), FP7 Marie Curie IOF (C.B.), Weatherstone Foundation fellowship (K.L.N.), National Institutes of Health K12 HD001259, the Hartwell Foundation (L.C.L., F.S.B.), the Lookout Fund and the McDonnell Foundation (F.H.G.). L.S.B.G. is an investigator with the Howard Hughes Medical Institute. Some experiments were conducted in J.F. Loring's laboratory (The Scripps Research Institute) with support from grants TR1-01250, CL1-00502, RM1-01717 (CIRM) and a gift from the Esther O'Keefe Foundation.

Author information

Affiliations

  1. Howard Hughes Medical Institute and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA

    • Mason A. Israel
    • , Shauna H. Yuan
    • , Sol M. Reyna
    • , Cheryl Herrera
    •  & Lawrence S. B. Goldstein
  2. Biomedical Sciences Graduate Program, University of California, San Diego, La Jolla, California 92093, USA

    • Mason A. Israel
    •  & Sol M. Reyna
  3. Department of Neurosciences, University of California, San Diego, La Jolla, California, USA

    • Shauna H. Yuan
    • , Edward H. Koo
    •  & Lawrence S. B. Goldstein
  4. The Salk Institute for Biological Studies, La Jolla, California 92037, USA

    • Cedric Bardy
    • , Yangling Mu
    •  & Fred H. Gage
  5. Department of Anesthesiology, University of California, San Diego, La Jolla, California 92093, USA

    • Michael P. Hefferan
    •  & Martin Marsala
  6. Department of Anesthesiology, Maastricht University Medical Center, Maastricht 6202 AZ, Netherlands

    • Sebastiaan Van Gorp
  7. Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA

    • Kristopher L. Nazor
  8. Department of Reproductive Medicine, University of California, San Diego, La Jolla, California 92093, USA

    • Francesca S. Boscolo
    •  & Louise C. Laurent
  9. BD Biosciences, La Jolla, California 92037, USA

    • Christian T. Carson
  10. Institute of Neurobiology, Slovak Academy of Sciences, Kosice SK-04001, Slovakia

    • Martin Marsala
  11. Department of Clinical Medicine, Neurology and Clinical Research Center, University of Oulu, Oulu FIN-90015, Finland

    • Anne M. Remes

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Contributions

M.A.I. and L.S.B.G. conceived the project; M.A.I. and L.S.B.G. designed the experiments; M.A.I., S.H.Y., C.B., S.M.R., Y.M., C.H., M.P.H., S.V.G., M.M., K.L.N. and F.S.B. performed the experiments; M.A.I., S.H.Y. and C.T.C. developed differentiation methods; A.M.R. and E.H.K. provided APPDp patient samples and information; F.H.G. supervised C.B. and Y.M.; M.M. supervised M.P.H. and S.V.G.; L.C.L. supervised K.L.N. and F.S.B.; M.A.I. and L.S.B.G. wrote the manuscript; F.H.G., E.H.K. and A.M.R. edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Lawrence S. B. Goldstein.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-12 with legends and Supplementary Tables 1, 3 and 4 (see separate files for Supplementary Tables 2 and 5).

  2. 2.

    Supplementary Table 5

    This table contains Amyloid-β, aGSK3β and p-tau/total tau detailed by patient, iPSC line and culture well.

Excel files

  1. 1.

    Supplementary Table 2

    This table shows differentially expressed genes between purified neurons and fetal brain versus non-neural fetal samples (sheet 1) and differentially expressed genes between purified neurons versus fetal brain (sheet 2).

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DOI

https://doi.org/10.1038/nature10821

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