Article | Published:

In vivo detection of amyloid-β deposits by near-infrared imaging using an oxazine-derivative probe

Nature Biotechnologyvolume 23pages577583 (2005) | Download Citation

Subjects

Abstract

As Alzheimer's disease pathogenesis is associated with the formation of insoluble aggregates of amyloid β-peptide, approaches allowing the direct, noninvasive visualization of plaque growth in vivo would be beneficial for biomedical research. Here we describe the synthesis and characterization of the near-infrared fluorescence oxazine dye AOI987, which readily penetrates the intact blood-brain barrier and binds to amyloid plaques. Using near-infrared fluorescence imaging, we demonstrated specific interaction of AOI987 with amyloid plaques in APP23 transgenic mice in vivo, as confirmed by postmortem analysis of brain slices. Quantitative analysis revealed increasing fluorescence signal intensity with increasing plaque load of the animals, and significant binding of AOI987 was observed for APP23 transgenic mice aged 9 months and older. Thus, AOI987 is an attractive probe to noninvasively monitor disease progression in animal models of Alzheimer disease and to evaluate effects of potential Alzheimer disease drugs on the plaque load.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Meyer-Luehmann, M. et al. Extracellular amyloid formation and associated pathology in neuronal grafts. Nat. Neurosci. 6, 1–8 (2003).

  2. 2

    Nussbaum, R.L. & Ellis, C.E. Alzheimer's disease and Parkinson's disease. N. Engl. J. Med. 348, 1356–1364 (2003).

  3. 3

    Hardy, J. & Selkoe, D.J. The amyloid hypothesis of Alzheimer's disease: progress and problems on the road to therapeutics. Science 297, 353–356 (2002).

  4. 4

    Zamrini, E., De Santi, S. & Tolar, M. Imaging is superior to cognitive testing for early diagnosis of Alzheimer's disease. Neurobiol. Aging 25, 685–691 (2004).

  5. 5

    Benveniste, H., Einstein, G., Kim, K.R., Hulette, C. & Johnson, G.A. Detection of neuritic plaques in Alzheimer's disease by magnetic resonance microscopy. Proc. Natl. Acad. Sci. USA 96, 14079–14084 (1999).

  6. 6

    Zhang, J. et al. Detection of amyloid plaques in mouse models of Alzheimer's disease by magnetic resonance imaging. Magn. Reson. Med. 51, 452–457 (2004).

  7. 7

    Mathis, C.A., Wang, Y. & Klunk, W.E. Imaging β-amyloid plaques and neurofibrillary tangles in the aging human brain. Curr. Pharm. Des. 10, 1469–1492 (2004).

  8. 8

    Bacskai, B.J., Klunk, W.E., Mathis, C.A. & Hyman, B.T. Imaging amyloid-β deposits in vivo. J. Cereb. Blood Flow Metab. 22, 1035–1041 (2002).

  9. 9

    Ono, M. et al. 11C-labeled stilbene derivatives as Aβ-aggregate-specific PET imaging agents for Alzheimer's disease. Nucl. Med. Biol. 30, 565–571 (2003).

  10. 10

    Suemoto, T. et al. In vivo labeling of amyloid with BF-108. Neurosci. Res. 48, 65–74 (2004).

  11. 11

    Okamura, N. et al. Styrylbenzoxazole derivatives for in vivo imaging of amyloid plaques in the brain. J. Neurosci. 24, 2535–2541 (2004).

  12. 12

    Klunk, W.E. et al. Uncharged thioflavin-T derivatives bind to amyloid-beta protein with high affinity and readily enter the brain. Life Sci. 69, 1471–1484 (2001).

  13. 13

    Mathis, C.A. et al. Synthesis and evaluation of 11C-labeled 6-substituted 2-arylbenzothiazoles as amyloid imaging agents. J. Med. Chem. 46, 2740–2754 (2003).

  14. 14

    Mathis, C.A. et al. A lipophilic thioflavin-T derivative for positron emission tomography (PET) imaging of amyloid in brain. Bioorg. Med. Chem. Lett. 12, 295–298 (2002).

  15. 15

    Agdeppa, E.D. et al. Binding characteristics of radiofluorinated 6-dialkylamino-2-naphthylethylidene derivatives as positron emission tomography imaging probes for β-amyloid plaques in Alzheimer's disease. J. Neurosci. 21, RC189 1–5 (2001).

  16. 16

    Agdeppa, E.D. et al. 2-Dialkylamino-6-acylmalononitrile substituted naphthalenes (DDNP analogs): novel diagnostic and therapeutic tools in Alzheimer's disease. Mol. Imaging Biol. 5, 404–417 (2003).

  17. 17

    Wadghiri, Y.Z. et al. Detection of Alzheimer's amyloid in transgenic mice using magnetic resonance microimaging. Magn. Reson. Med. 50, 293–302 (2003).

  18. 18

    Poduslo, J.F. et al. Design and chemical synthesis of a magnetic resonance contrast agent with enhanced in vitro binding, high blood-brain barrier permeability, and in vivo targeting to Alzheimer's disease amyloid plaques. Biochemistry 43, 6064–6075 (2004).

  19. 19

    Poduslo, J.F. et al. Molecular targeting of Alzheimer's amyloid plaques for contrast-enhanced magnetic resonance imaging. Neurobiol. Dis. 11, 315–329 (2002).

  20. 20

    Bacskai, B.J. et al. Imaging of amyloid-β deposits in brains of living mice permits direct observation of clearance of plaques with immunotherapy. Nat. Med. 7, 369–372 (2001).

  21. 21

    Klunk, W. et al. Imaging Aβ plaques in living transgenic mice with multiphoton microscopy and methoxy-X04, a systemically administered Congo red derivative. J. Neuropathol. Exp. Neurol. 61, 797–805 (2002).

  22. 22

    Bacskai, B.J. et al. Four-dimensional multiphoton imaging of brain entry, amyloid binding, and clearance of an amyloid-β ligand in transgenic mice. Proc. Natl. Acad. Sci. USA 100, 12462–12467 (2003).

  23. 23

    Styren, S.D., Hamilton, R.L., Styren, G.C. & Klunk, W.E. X-34, a fluorescent derivative of Congo red: a novel histochemical stain for Alzheimer's disease pathology. J. Histochem. Cytochem. 48, 1223–1232 (2000).

  24. 24

    Shoghi-Jadid, K. et al. Localization of neurofibrillary tangles and beta-amyloid plaques in the brains of living patients with Alzheimer disease. Am. J. Geriatr. Psychiatry 10, 24–35 (2002).

  25. 25

    Klunk, W.E. et al. Imaging brain amyloid in Alzheimer's disease with Pittsburgh compound-B. Ann. Neurol. 55, 306–319 (2004).

  26. 26

    Weissleder, R. & Ntziachristos, V. Shedding light onto live molecular targets. Nat. Med. 9, 123–128 (2003).

  27. 27

    Ntziachristos, V., Bremer, C. & Weissleder, R. Fluorescence imaging with near-infrared light: new technological advances that enable in vivo molecular imaging. Eur. Radiol. 13, 195–208 (2003).

  28. 28

    Funovics, M., Weissleder, R. & Tung, C.-H. Protease sensors for bioimaging. Anal. Bioanal. Chem. 377, 956–963 (2003).

  29. 29

    Sturchler-Pierrat, C. et al. Two amyloid precursor protein transgenic mouse models with Alzheimer disease-like pathology. Proc. Natl. Acad. Sci. USA 94, 13287–13292 (1997).

  30. 30

    Neuweiler, H. et al. Detection of individual p53-autoantibodies by using quenched peptide-based molecular probes. Angew. Chem. Int. Edn. Engl. 41, 4769–4773 (2002).

  31. 31

    Bremer, C., Tung, C.H. & Weissleder, R. In vivo molecular target assessment of matrix metalloproteinase inhibition. Nat. Med. 7, 743–748 (2001).

  32. 32

    Wunder, A. et al. In vivo imaging of protease activity in arthritis: a novel approach for monitoring treatment response. Arthritis Rheum. 50, 2459–2465 (2004).

  33. 33

    Graves, E.E. et al. A submillimeter resolution fluorescence molecular imaging system for small animal imaging. Med. Phys. 30, 901–911 (2003).

  34. 34

    Rudin, M. & Weissleder, R. Molecular imaging in drug discovery and development. Nat. Rev. Drug Discov. 2, 123–131 (2003).

  35. 35

    Daves, G. & McMurtrey, K.D. Koenig's adducts of N-alkyl(aryl)-aminoethanols and quinones. 3,4-Dihydro-4-alkyl(aryl)-8a-hydroxy-2H–1,4-benzoxazine-6(8aH)-ones. J. Org. Chem. 35, 4252–4253 (1970).

  36. 36

    Klunk, W.E. & Pettegrew, J.W. Development of small molecule probes for the beta-amyloid protein of Alzheimer's disease. Neurobiol. Aging 15, 691–698 (1994).

  37. 37

    Levine, H. III Thioflavin T interaction with synthetic Alzheimer's disease β-amyloid peptides: detection of amyloid aggregation in solution. Protein Sci. 2, 404–410 (1993).

Download references

Acknowledgements

The authors would like to thank Alexandra Suter for her excellent technical assistance.

Author information

Affiliations

  1. Discovery Technologies, Novartis Institutes for Biomedical Research, Basel, CH-4002, Switzerland

    • Martin Hintersteiner
    • , Willy Kinzy
    • , Rainer Kneuer
    • , Markus Rudin
    • , Markus Stoeckli
    •  & Hans-Ulrich Gremlich
  2. Nervous System Department, Novartis Institutes for Biomedical Research, Basel, CH-4002, Switzerland

    • Albert Enz
    • , Peter Frey
    • , Anne-Lise Jaton
    • , Ulf Neumann
    • , Matthias Staufenbiel
    •  & Karl-Heinz Wiederhold

Authors

  1. Search for Martin Hintersteiner in:

  2. Search for Albert Enz in:

  3. Search for Peter Frey in:

  4. Search for Anne-Lise Jaton in:

  5. Search for Willy Kinzy in:

  6. Search for Rainer Kneuer in:

  7. Search for Ulf Neumann in:

  8. Search for Markus Rudin in:

  9. Search for Matthias Staufenbiel in:

  10. Search for Markus Stoeckli in:

  11. Search for Karl-Heinz Wiederhold in:

  12. Search for Hans-Ulrich Gremlich in:

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Hans-Ulrich Gremlich.

About this article

Publication history

Received

Accepted

Published

Issue Date

DOI

https://doi.org/10.1038/nbt1085

Further reading