Abstract

Alzheimer’s disease is a neurodegenerative disorder associated with the aberrant aggregation of the amyloid-β peptide. Although increasing evidence implicates cholesterol in the pathogenesis of Alzheimer’s disease, the detailed mechanistic link between this lipid molecule and the disease process remains to be fully established. To address this problem, we adopt a kinetics-based strategy that reveals a specific catalytic role of cholesterol in the aggregation of Aβ42 (the 42-residue form of the amyloid-β peptide). More specifically, we demonstrate that lipid membranes containing cholesterol promote Aβ42 aggregation by enhancing its primary nucleation rate by up to 20-fold through a heterogeneous nucleation pathway. We further show that this process occurs as a result of cooperativity in the interaction of multiple cholesterol molecules with Aβ42. These results identify a specific microscopic pathway by which cholesterol dramatically enhances the onset of Aβ42 aggregation, thereby helping rationalize the link between Alzheimer’s disease and the impairment of cholesterol homeostasis.

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References

  1. 1.

    Alzheimer’s Association. 2012 Alzheimer’s disease facts and figures. Alzheimer’s Dement 8, 131–168 (2012).

  2. 2.

    Dobson, C. M. Protein folding and misfolding. Nature 426, 884–890 (2003).

  3. 3.

    Haass, C. & Selkoe, D. J. Soluble protein oligomers in neurodegeneration: lessons from the Alzheimer’s amyloid beta-peptide. Nat. Rev. Mol. Cell Biol. 8, 101–112 (2007).

  4. 4.

    Knowles, T. P. J., Vendruscolo, M. & Dobson, C. M. The amyloid state and its association with protein misfolding diseases. Nat. Rev. Mol. Cell Biol. 15, 384–396 (2014).

  5. 5.

    Tanzi, R. E. & Bertram, L. Twenty years of the Alzheimer’s disease amyloid hypothesis: a genetic perspective. Cell 120, 545–555 (2005).

  6. 6.

    Necula, M., Kayed, R., Milton, S. & Glabe, C. G. Small molecule inhibitors of aggregation indicate that amyloid β oligomerization and fibrillization pathways are independent and distinct. J. Biol. Chem. 282, 10311–10324 (2007).

  7. 7.

    Lansbury, P. T. & Lashuel, H. A. A century-old debate on protein aggregation and neurodegeneration enters the clinic. Nature 443, 774–779 (2006).

  8. 8.

    Galvagnion, C. et al. Lipid vesicles trigger α-synuclein aggregation by stimulating primary nucleation. Nat. Chem. Biol. 11, 229–234 (2015).

  9. 9.

    Di Paolo, G. & Kim, T.-W. Linking lipids to Alzheimer’s disease: cholesterol and beyond. Nat. Rev. Neurosci. 12, 284–296 (2011).

  10. 10.

    Gellermann, G. P. et al. Raft lipids as common components of human extracellular amyloid fibrils. Proc. Natl Acad. Sci. USA 102, 6297–6302 (2005).

  11. 11.

    Bertram, L. & Tanzi, R. E. Thirty years of Alzheimer’s disease genetics: the implications of systematic meta-analyses. Nat. Rev. Neurosci. 9, 768–778 (2008).

  12. 12.

    Corder, E. H. et al. Gene dose of apolipoprotein E type 4 allele and the risk of Alzheimer’s disease in late onset families. Science 261, 921–923 (1993).

  13. 13.

    Bu, G. Apolipoprotein E and its receptors in Alzheimer’s disease: pathways, pathogenesis and therapy. Nat. Rev. Neurosci. 10, 333–344 (2009).

  14. 14.

    Holtzman, D. M. Role of apoE/Aβ interactions in the pathogenesis of Alzheimer’s disease and cerebral amyloid angiopathy. J. Mol. Neurosci. 17, 147–155 (2001).

  15. 15.

    Popp, J. et al. Cerebral and extracerebral cholesterol metabolism and CSF markers of Alzheimer’s disease. Biochem. Pharmacol. 86, 37–42 (2013).

  16. 16.

    Mori, T. et al. Cholesterol accumulates in senile plaques of Alzheimer disease patients and in transgenic APP(SW) mice. J. Neuropathol. Exp. Neurol. 60, 778–785 (2001).

  17. 17.

    Zissimopoulos, J. M. et al. Sex and race differences in the association between statin use and the incidence of Alzheimer disease. JAMA Neurol. 111, 390–400 (2016).

  18. 18.

    Dietschy, J. M. & Turley, S. D. Cholesterol metabolism in the brain. Curr. Opin. Lipidol. 12, 105–112 (2001).

  19. 19.

    Vance, J. E. Dysregulation of cholesterol balance in the brain: contribution to neurodegenerative diseases. Dis. Model. Mech. 5, 746–755 (2012).

  20. 20.

    Wood, W. G., Li, L., Müller, W. E. & Eckert, G. P. Cholesterol as a causative agent in Alzheimer disease a debatable hypothesis. J. Neurochem. 129, 559–572 (2014).

  21. 21.

    Di Scala, C., Chahinian, H., Yahi, N., Garmy, N. & Fantini, J. Interaction of Alzheimer’s β-amyloid peptides with cholesterol: mechanistic insights into amyloid pore formation. Biochemistry 53, 4489–4502 (2014).

  22. 22.

    Michaels, T. C. T., Lazell, H. W., Arosio, P. & Knowles, T. P. J. Dynamics of protein aggregation and oligomer formation governed by secondary nucleation. J. Chem. Phys. 143, 54901 (2015).

  23. 23.

    Arosio, P., Vendruscolo, M., Dobson, C. M. & Knowles, T. P. J. Chemical kinetics for drug discovery to combat protein aggregation diseases. Trends Pharmacol. Sci. 35, 127–135 (2014).

  24. 24.

    Ghribi, O., Larsen, B., Schrag, M. & Herman, M. M. High cholesterol content in neurons increases BACE, β-amyloid, and phosphorylated tau levels in rabbit hippocampus. Exp. Neurol. 200, 460–467 (2006).

  25. 25.

    Barrett, P. J. et al. The amyloid precursor protein has a flexible transmembrane domain and binds cholesterol. Science 117, 2010–2013 (2012).

  26. 26.

    Evangelisti, E. et al. Membrane lipid composition and its physicochemical properties define cell vulnerability to aberrant protein oligomers. J. Cell Sci. 125, 2416–2427 (2012).

  27. 27.

    Hellstrand, E., Sparr, E. & Linse, S. Retardation of Aβ fibril formation by phospholipid vesicles depends on membrane phase behavior. Biophys. J. 98, 2206–2214 (2010).

  28. 28.

    Yip, C. M., Elton, E. A Darabie, A. A., Morrison, M. R. & McLaurin, J. Cholesterol, a modulator of membrane-associated A beta-fibrillogenesis and neurotoxicity. J. Mol. Biol. 311,723–734 (2001).

  29. 29.

    Hellstrand, E., Boland, B., Walsh, D. M. & Linse, S. Amyloid β-protein aggregation produces highly reproducible kinetic data and occurs by a two-phase process. ACS Chem. Neurosci. 1, 13–18 (2010).

  30. 30.

    Knowles, T. P. J. et al. An analytical solution to the kinetics of breakable filament assembly. Science 326, 1533–1537 (2009).

  31. 31.

    Cohen, S. I. A. et al. Proliferation of amyloid-β42 aggregates occurs through a secondary nucleation mechanism. Proc. Natl Acad. Sci. USA 110, 9758–9763 (2013).

  32. 32.

    Cohen, S. I. A., Vendruscolo, M., Dobson, C. M. & Knowles, T. P. J. From macroscopic measurements to microscopic mechanisms of protein aggregation. J. Mol. Biol. 421, 160–171 (2012).

  33. 33.

    Meisl, G. et al. Molecular mechanisms of protein aggregation from global fitting of kinetic models. Nat. Protoc. 11, 252–272 (2016).

  34. 34.

    Meisl, G., Yang, X., Frohm, B., Knowles, T. P. J. & Linse, S. Quantitative analysis of intrinsic and extrinsic factors in the aggregation mechanism of Alzheimer-associated Aβ-peptide. Sci. Rep. 6, 18728 (2016).

  35. 35.

    Meisl, G. et al. Differences in nucleation behavior underlie the contrasting aggregation kinetics of the Aβ40 and Aβ42 peptides. Proc. Natl Acad. Sci. USA 111, 9384–9389 (2014).

  36. 36.

    Arosio, P. et al. Kinetic analysis reveals the diversity of microscopic mechanisms through which molecular chaperones suppress amyloid formation. Nat. Commun. 7, 10948 (2016).

  37. 37.

    Cohen, S. I. A. et al. A molecular chaperone breaks the catalytic cycle that generates toxic Aβ oligomers. Nat. Struct. Mol. Biol. 22, 207–213 (2015).

  38. 38.

    Sormanni, P., Aprile, F. A. & Vendruscolo, M. Rational design of antibodies targeting specific epitopes within intrinsically disordered proteins. Proc. Natl Acad. Sci. USA 112, 9902–9907 (2015).

  39. 39.

    Habchi, J. et al. An anti-cancer drug suppresses the primary nucleation reaction that initiates the formation of toxic Aβ aggregates associated with Alzheimer’s disease. Sci. Adv. 2, e1501244 (2016).

  40. 40.

    Habchi, J. et al. Systematic development of small molecules to inhibit specific microscopic steps of Aβ42 aggregation in Alzheimer’s disease. Proc. Natl Acad. Sci. USA 114, E200–E208 (2016).

  41. 41.

    Sastry, P. S. Lipids of nervous tissue: composition and metabolism. Prog. Lipid Res. 24, 69–176 (1985).

  42. 42.

    van Meer, G., Voelker, D. R. & Feigenson, G. W. Membrane lipids: where they are and how they behave. Nat. Rev. Mol. Cell Biol. 9, 112–124 (2008).

  43. 43.

    Mouritsen, O. G. Life—As a Matter of Fat (Springer, Berlin, 2005).

  44. 44.

    Cotman, C. W., Blank, M., Moehl, A. & Snyder, F. Lipid composition of synaptic plasma membranes isolated from rat brain by zonal centrifugation. Biochemistry 8, 4606–4612 (1969).

  45. 45.

    Nagle, J. F. et al. X-ray structure determination of fully hydrated L alpha phase dipalmitoylphosphatidylcholine bilayers. Biophys. J. 70, 1419–1431 (1996).

  46. 46.

    Kucerka, N. et al. Structure of fully hydrated fluid phase DMPC and DLPC lipid bilayers using X-ray scattering from oriented multilamellar arrays and from unilamellar vesicles. Biophys. J. 88, 2626–2637 (2005).

  47. 47.

    Chi, E. Y. et al. Lipid membrane templates the ordering and induces the fibrillogenesis of Alzheimer’s disease amyloid-β peptide. Prot. Struct. Funct. Genet. 72, 1–24 (2008).

  48. 48.

    Niu, Z. et al. The molecular structure of Alzheimer β-amyloid fibrils formed in the presence of phospholipid vesicles. Angew. Chem. Int. Ed. 53, 9294–9297 (2014).

  49. 49.

    Simons, K. & Vaz, W. L. C. Model systems, lipid rafts, and cell membranes. Annu. Rev. Biophys. Biomol. Struct. 33, 269–295 (2004).

  50. 50.

    Filippov, A., Orädd, G. & Lindblom, G. The effect of cholesterol on the lateral diffusion of phospholipids in oriented bilayers. Biophys. J. 84, 3079–3086 (2003).

  51. 51.

    Barrett, M. A. et al. Solubility of cholesterol in lipid membranes and the formation of immiscible cholesterol plaques at high cholesterol concentrations. Soft Matter 9, 9342–9351 (2013).

  52. 52.

    Almeida, P. F., Vaz, W. L. & Thompson, T. E. Lateral diffusion in the liquid phases of dimyristoylphosphatidylcholine/cholesterol lipid bilayers: a free volume analysis. Biochemistry 31, 6739–6747 (1992).

  53. 53.

    Blume, A. A comparative study of the phase transitions of phospholipid bilayers and monolayers. Biochim. Biophys. Acta Biomembr. 557, 32–44 (1979).

  54. 54.

    Wimley, W. C. & Thompson, T. E. Transbilayer and interbilayer phospholipid exchange in dimyristoylphosphatidylcholine/dimyristoylphosphatidylethanolamine large unilamellar vesicles. Biochemistry 30, 1702–1709 (1991).

  55. 55.

    Harris, F. M., Best, K. B. & Bell, J. D. Use of laurdan fluorescence intensity and polarization to distinguish between changes in membrane fluidity and phospholipid order. Biochim. Biophys. Acta Biomembr. 1565, 123–128 (2002).

  56. 56.

    Aguilar, L. F. et al. Differential dynamic and structural behavior of lipid-cholesterol domains in model membranes. PLoS One 7, e40254 (2012).

  57. 57.

    Galvagnion, C. et al. Chemical properties of lipids strongly affect the kinetics of the membrane-induced aggregation of α-synuclein. Proc. Natl Acad. Sci. USA 113, 7065–7070 (2016).

  58. 58.

    De Meyer, F. & Smit, B. Effect of cholesterol on the structure of a phospholipid bilayer. Proc. Natl Acad. Sci. USA 106, 3654–3658 (2009).

  59. 59.

    de Jongh, H. H. J., Goormaghtigh, E. & Killian, J. A. Analysis of circular dichroism spectra of oriented protein–lipid complexes: toward a general application. Biochemistry 33, 14521–14528 (1994).

  60. 60.

    Linse, S. & Lund, M. Surface effects on aggregation kinetics of amyloidogenic peptides. J. Am. Chem. Soc. 136, 11555–11850 (2014).

  61. 61.

    Ruggeri, F. S. et al. Nanoscale studies link amyloid maturity with polyglutamine diseases onset. Sci. Rep. 6, 31155 (2016).

  62. 62.

    Zandomeneghi, G., Krebs, M. R. H., McCammon, M. G. & Fändrich, M. FTIR reveals structural differences between native beta-sheet proteins and amyloid fibrils. Protein Sci. 13, 3314–3321 (2004).

  63. 63.

    Arosio, P., Knowles, T. P. J. & Linse, S. On the lag phase in amyloid fibril formation. Phys. Chem. Chem. Phys. 17, 7606–7618 (2015).

  64. 64.

    Cohen, S. I. A., Vendruscolo, M., Dobson, C. M. & Knowles, T. P. J. Nucleated polymerization with secondary pathways. II. Determination of self-consistent solutions to growth processes described by non-linear master equations. J. Chem. Phys. 135, 65106 (2011).

  65. 65.

    Cohen, S. I. A., Vendruscolo, M., Dobson, C. M. & Knowles, T. P. J. Nucleated polymerization with secondary pathways. III. Equilibrium behavior and oligomer populations. J. Chem. Phys. 135, 65107 (2011).

  66. 66.

    Cohen, S. I. A. et al. Nucleated polymerization with secondary pathways. I. Time evolution of the principal moments. J. Chem. Phys. 135, 65105 (2011).

  67. 67.

    Simons, M. et al. Cholesterol depletion inhibits the generation of β-amyloid in hippocampal neurons. Proc. Natl Acad. Sci. USA 95, 6460–6464 (1998).

  68. 68.

    Hong, S. et al. Soluble Aβ oligomers are rapidly sequestered from brain ISF in vivo and bind GM1 ganglioside on cellular membranes. Neuron 82, 308–319 (2014).

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Acknowledgements

The authors acknowledge support from the Centre for Misfolding Diseases (J.H., S.C., T.C.T.M., M.M.J.B., F.S.R., M.S., J.R.K., C.M.D., T.P.J.K. and M.V.); the Agency for Science, Technology and Research, Singapore (S.C.); a Marie Skłodowska-Curie Actions — Individual Fellowship (C.G.); Peterhouse College, Cambridge (T.C.T.M.); the Swiss National Science Foundation (T.C.T.M., F.S.R.); the NIH-Oxford/Cambridge Scholars Program (M.M.J.B.); the Cambridge Commonwealth, European and International Trust (M.M.J.B.); the Knut & Alice Wallenberg Foundation (S.L., E.S.); the European Research Council (S.L.); the Swedish Research Council (S.L., E.S.) the Frances and Augustus Newman Foundation (T.P.J.K.); the UK Biotechnology and Biochemical Sciences Research Council (C.M.D. and M.V.); and the Wellcome Trust (C.M.D., T.P.J.K. and M.V.). This work was supported by the Intramural Research Program of the National Institute of Diabetes and Kidney Diseases, NIH.

Author information

Author notes

    • Céline Galvagnion

    Present address: German Center for Neurodegenerative Diseases (DZNE), Bonn, Germany

  1. These authors contributed equally: Johnny Habchi, Sean Chia, Céline Galvagnion.

Affiliations

  1. Centre for Misfolding Diseases, Department of Chemistry, University of Cambridge, Cambridge, UK

    • Johnny Habchi
    • , Sean Chia
    • , Céline Galvagnion
    • , Thomas C. T. Michaels
    • , Mathias M. J. Bellaiche
    • , Francesco Simone Ruggeri
    • , Michele Sanguanini
    • , Janet R. Kumita
    • , Christopher M. Dobson
    • , Tuomas P. J. Knowles
    •  & Michele Vendruscolo
  2. Paulson School for Engineering and Applied Sciences, Harvard University, Cambridge, USA

    • Thomas C. T. Michaels
  3. Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA

    • Mathias M. J. Bellaiche
  4. Department of Biochemistry & Structural Biology, Center for Molecular Protein Science, Lund University, Lund, Sweden

    • Ilaria Idini
    •  & Sara Linse
  5. Division of Physical Chemistry, Department of Chemistry, Lund University, Lund, Sweden

    • Emma Sparr
  6. Department of Physics, Cavendish Laboratory, Cambridge, UK

    • Tuomas P. J. Knowles

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Contributions

J.H., S.C., C.G., T.C.T.M., E.S., S.L., C.M.D., T.P.J.K. and M.V. designed the research. J.H., S.C., C.G., F.S.R., M.S. and I.I. performed the research. J.H., S.C., C.G., F.S.R., I.I., J.R.K., E.S., S.L., C.M.D., T.P.J.K. and M.V. contributed reagents/analytic tools. J.H., S.C., C.G., T.C.T.M., M.M.J.B., F.S.R., E.S., S.L., C.M.D., T.P.J.K. and M.V. analysed the data. All authors discussed the results and contributed to the writing of the paper.

Competing interests

The authors declare no competing interests.

Corresponding author

Correspondence to Michele Vendruscolo.

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https://doi.org/10.1038/s41557-018-0031-x

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