Letter

A three-dimensional human neural cell culture model of Alzheimer’s disease

  • Nature volume 515, pages 274278 (13 November 2014)
  • doi:10.1038/nature13800
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Abstract

Alzheimer’s disease is the most common form of dementia, characterized by two pathological hallmarks: amyloid-β plaques and neurofibrillary tangles1. The amyloid hypothesis of Alzheimer’s disease posits that the excessive accumulation of amyloid-β peptide leads to neurofibrillary tangles composed of aggregated hyperphosphorylated tau2,3. However, to date, no single disease model has serially linked these two pathological events using human neuronal cells. Mouse models with familial Alzheimer’s disease (FAD) mutations exhibit amyloid-β-induced synaptic and memory deficits but they do not fully recapitulate other key pathological events of Alzheimer’s disease, including distinct neurofibrillary tangle pathology4,5. Human neurons derived from Alzheimer’s disease patients have shown elevated levels of toxic amyloid-β species and phosphorylated tau but did not demonstrate amyloid-β plaques or neurofibrillary tangles6,7,8,9,10,11. Here we report that FAD mutations in β-amyloid precursor protein and presenilin 1 are able to induce robust extracellular deposition of amyloid-β, including amyloid-β plaques, in a human neural stem-cell-derived three-dimensional (3D) culture system. More importantly, the 3D-differentiated neuronal cells expressing FAD mutations exhibited high levels of detergent-resistant, silver-positive aggregates of phosphorylated tau in the soma and neurites, as well as filamentous tau, as detected by immunoelectron microscopy. Inhibition of amyloid-β generation with β- or γ-secretase inhibitors not only decreased amyloid-β pathology, but also attenuated tauopathy. We also found that glycogen synthase kinase 3 (GSK3) regulated amyloid-β-mediated tau phosphorylation. We have successfully recapitulated amyloid-β and tau pathology in a single 3D human neural cell culture system. Our unique strategy for recapitulating Alzheimer’s disease pathology in a 3D neural cell culture model should also serve to facilitate the development of more precise human neural cell models of other neurodegenerative disorders.

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Acknowledgements

This work was supported by the grants from the Cure Alzheimer’s fund (D.Y.K., S.H.C. and R.E.T.) and national Institute of Health grants 5P01AG15379 (R.E.T.) and 5R37MH060009 (R.E.T.). We thank T. L. Spires, M. Polydoro and S. Wegmann for revising the manuscript, and M. L. McKee for the electron microscopy assistance. We also appreciate B. T. Hyman, O. Berezovska, J. Hardy and P. Davies for providing cDNAs and antibodies. We acknowledge Ragon Institute’s Imaging Core facility (part of the Harvard CFAR Immunology Core), Massachusetts General Hospital (MGH) Viral Vector Core (supported by NIH/NINDS P30NS04776), MGH Microscopy Core of the Center for Systems Biology for immunoelectron microscopy (partially supported by an IBDG Grant DK43351 and a BADERC Award DK57521), MGH Confocal Microscope Core and MGH Pathology Core for technical and instrument support.

Author information

Author notes

    • Se Hoon Choi
    •  & Young Hye Kim

    These authors contributed equally to this work.

Affiliations

  1. Genetics and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts 02129, USA

    • Se Hoon Choi
    • , Young Hye Kim
    • , Matthias Hebisch
    • , Christopher Sliwinski
    • , Carla D’Avanzo
    • , Hechao Chen
    • , Basavaraj Hooli
    • , Caroline Asselin
    • , Justin B. Klee
    • , Can Zhang
    • , Dora M. Kovacs
    • , Rudolph E. Tanzi
    •  & Doo Yeon Kim
  2. Division of Mass Spectrometry Research, Korea Basic Science Institute, Cheongju-si, Chungbuk 363-883, South Korea

    • Young Hye Kim
  3. Institute of Reconstructive Neurobiology, Life and Brain Center, University of Bonn and Hertie Foundation, 53127 Bonn, Germany

    • Matthias Hebisch
    •  & Michael Peitz
  4. FM Kirby Neurobiology Center, Boston Children’s Hospital and Harvard Stem Cell Institute, Boston, Massachusetts 02115, USA

    • Seungkyu Lee
    • , Brian J. Wainger
    •  & Clifford J. Woolf
  5. The Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA

    • Julien Muffat
  6. Department of Neurosciences, University of California, San Diego, La Jolla, California 92093, USA

    • Steven L. Wagner

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Contributions

D.Y.K. and R.E.T. were equally responsible for experimental design and data interpretation. S.H.C., Y.H.K. and D.Y.K. mainly contributed to writing and revising the manuscript. D.Y.K., Y.H.K., S.H.C., M.H., S.L., C.D., H.C., C.S., B.H., J.B.K., C.A. and C.Z. conducted the experiments. S.L.W. synthesized SGSM41 and S.L.W. and C.Z. characterized SGSM41. J.M., B.J.W., M.P., C.J.W. and D.M.K. contributed to data interpretation.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Rudolph E. Tanzi or Doo Yeon Kim.

Extended data

Supplementary information

Videos

  1. 1.

    3D reconstitution of ReN-G cells after 2-week differentiation in 3D Matrigel (green, GFP; 400x magnification)

    Due to the technical limitation of spinning disk confocal microscopy (Olympus), 3D reconstituted images represent only a part of the whole 3D culture.

  2. 2.

    3D reconstitution of ReN-m (enriched) cells after 4-week differentiation in 3D Matrigel (red, mCherry; 400x magnification)

    3D reconstitution of ReN-m (enriched) cells after 4-week differentiation in 3D Matrigel (red, mCherry; 400x magnification)

  3. 3.

    3D reconstitution of 3D-differentiated ReN-m cells for 6 weeks (green, Synapsin I; red, mCherry; 1,000x magnification).

    3D reconstitution of 3D-differentiated ReN-m cells for 6 weeks (green, Synapsin I; red, mCherry; 1,000x magnification).

  4. 4.

    Z-section sequences of 3D6-stained Aβ aggregates in 6-week differentiated ReN-mGAP cells (green, GFP; 400x magnification).

    Z-section sequences of 3D6-stained Aβ aggregates in 6-week differentiated ReN-mGAP cells (green, GFP; 400x magnification).

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