Top

Published in:

Open Access 01-12-2015 | Research

APLP1 as a cerebrospinal fluid biomarker for γ-secretase modulator treatment

Authors: Simon Sjödin, Kerstin K. A. Andersson, Marc Mercken, Henrik Zetterberg, Herman Borghys, Kaj Blennow, Erik Portelius

Published in: Alzheimer's Research & Therapy | Issue 1/2015

Abstract

Introduction

Alzheimer’s disease brains are characterized by extracellular plaques containing the aggregated amyloid β₄₂ (Aβ₄₂) peptide and intraneuronal tangles containing hyperphosphorylated tau. Aβ₄₂ is produced by sequential processing of the amyloid precursor protein (APP) by β-secretase followed by γ-secretase. Substantial efforts have been put into developing pharmaceuticals preventing the production or increasing the clearance of Aβ₄₂. However, treatments inhibiting γ-secretase have proven disappointing due to off-target effects. To circumvent these effects, γ-secretase modulators (GSMs) have been developed, which rather than inhibiting γ-secretase shift its preference into producing less aggregation-prone shorter Aβ peptides. Belonging to the same family of proteins as APP, amyloid-like protein 1 (APLP1) is also a substrate for γ-secretase. Herein we investigated whether the GSM E2012 affects APLP1 processing in the central nervous system by measuring APLP1 peptide levels in cerebrospinal fluid (CSF) before and after E2012 treatment in dogs.

Methods

An in-house monoclonal APLP1 antibody, AP1, was produced and utilized for immunopurification of APLP1 from human and dog CSF in a hybrid immuno-affinity mass spectrometric method. Seven dogs received a single dose of 20 or 80 mg/kg of E2012 in a randomized cross-over design and CSF was collected prior to and 4, 8 and 24 hours after dosing.

Results

We have identified 14 CSF APLP1 peptides in humans and 12 CSF APLP1 peptides in dogs. Of these, seven were reproducibly detectable in dogs who received E2012. We found a dose-dependent relative increase of the CSF peptides APLP1β17, 1β18 and 1β28 accompanied with a decrease of 1β25 and 1β27 in response to E2012 treatment. All peptides reverted to baseline over the time of sample collection.

Conclusion

We show an in vivo effect of the GSM E2012 on the processing of APLP1 which is measurable in CSF. These data suggest that APLP1 peptides may be used as biomarkers to monitor drug effects of GSMs on γ-secretase processing in clinical trials. However, this requires further investigation in larger cohorts, including studies in man.

Mayeux R, Stern Y. Epidemiology of Alzheimer disease. Cold Spring Harb Perspect Med. 2012;2:1–18. doi:10.1101/cshperspect.a006239.CrossRef

Masters CL, Simms G, Weinman NA, Multhaup G, McDonald BL, Beyreuther K. Amyloid plaque core protein in Alzheimer disease and Down syndrome. Proc Natl Acad Sci U S A. 1985;82:4245–9.PubMedCentralCrossRefPubMed

Grundke-Iqbal I, Iqbal K, Tung YC, Quinlan M, Wisniewski HM, Binder LI. Abnormal phosphorylation of the microtubule-associated protein tau (tau) in Alzheimer cytoskeletal pathology. Proc Natl Acad Sci U S A. 1986;83:4913–7.PubMedCentralCrossRefPubMed

Hardy JA, Higgins GA. Alzheimer’s disease: the amyloid cascade hypothesis. Science. 1992;256:184–5.CrossRefPubMed

De Strooper B. Lessons from a failed gamma-secretase Alzheimer trial. Cell. 2014;159:721–6. doi:10.1016/j.cell.2014.10.016.CrossRefPubMed

Mangialasche F, Solomon A, Winblad B, Mecocci P, Kivipelto M. Alzheimer’s disease: clinical trials and drug development. Lancet Neurol. 2010;9:702–16. http://dx.doi.org/10.1016/S1474-4422(10)70119-8 (accessed 2015-11-09).

Citron M. Alzheimer’s disease: strategies for disease modification. Nat Rev Drug Discov. 2010;9:387–98.CrossRefPubMed

Kuhn PH, Wang H, Dislich B, Colombo A, Zeitschel U, Ellwart JW, et al. ADAM10 is the physiologically relevant, constitutive alpha-secretase of the amyloid precursor protein in primary neurons. EMBO J. 2010;29:3020–32. doi:10.1038/emboj.2010.167.PubMedCentralCrossRefPubMed

Vassar R, Bennett BD, Babu-Khan S, Kahn S, Mendiaz EA, Denis P, et al. Beta-secretase cleavage of Alzheimer’s amyloid precursor protein by the transmembrane aspartic protease BACE. Science. 1999;286:735–41.CrossRefPubMed

10.

Kimberly WT, LaVoie MJ, Ostaszewski BL, Ye W, Wolfe MS, Selkoe DJ. Gamma-secretase is a membrane protein complex comprised of presenilin, nicastrin, Aph-1, and Pen-2. Proc Natl Acad Sci U S A. 2003;100:6382–7. doi:10.1073/pnas.1037392100.PubMedCentralCrossRefPubMed

11.

De Strooper B, Saftig P, Craessaerts K, Vanderstichele H, Guhde G, Annaert W, et al. Deficiency of presenilin-1 inhibits the normal cleavage of amyloid precursor protein. Nature. 1998;391:387–90. doi:10.1038/34910.CrossRefPubMed

12.

Esch FS, Keim PS, Beattie EC, Blacher RW, Culwell AR, Oltersdorf T, et al. Cleavage of amyloid beta peptide during constitutive processing of its precursor. Science. 1990;248:1122–4.CrossRefPubMed

13.

Doody RS, Raman R, Farlow M, Iwatsubo T, Vellas B, Joffe S, et al. A phase 3 trial of semagacestat for treatment of Alzheimer’s disease. N Engl J Med. 2013;369:341–50. doi:10.1056/NEJMoa1210951.CrossRefPubMed

14.

Coric V, van Dyck CH, Salloway S, Andreasen N, Brody M, Richter RW, et al. Safety and tolerability of the gamma-secretase inhibitor avagacestat in a phase 2 study of mild to moderate Alzheimer disease. Arch Neurol. 2012;69:1430–40. doi:10.1001/archneurol.2012.2194.CrossRefPubMed

15.

McCarthy JV, Twomey C, Wujek P. Presenilin-dependent regulated intramembrane proteolysis and γ-secretase activity. Cell Mol Life Sci. 2009;66:1534–55. doi:10.1007/s00018-009-8435-9.CrossRefPubMed

16.

De Strooper B, Annaert W, Cupers P, Saftig P, Craessaerts K, Mumm JS, et al. A presenilin-1-dependent gamma-secretase-like protease mediates release of Notch intracellular domain. Nature. 1999;398:518–22. doi:10.1038/19083.CrossRefPubMed

17.

Struhl G, Greenwald I. Presenilin is required for activity and nuclear access of Notch in Drosophila. Nature. 1999;398:522–5. doi:10.1038/19091.CrossRefPubMed

18.

Barthet G, Georgakopoulos A, Robakis NK. Cellular mechanisms of gamma-secretase substrate selection, processing and toxicity. Prog Neurobiol. 2012;98:166–75. doi:10.1016/j.pneurobio.2012.05.006.CrossRefPubMed

19.

Ling IF, Golde TE, Galasko DR, Koo EH. Modulation of Abeta42 in vivo by gamma-secretase modulator in primates and humans. Alzheimers Res Ther. 2015;7:55. doi:10.1186/s13195-015-0137-y.PubMedCentralCrossRefPubMed

20.

Portelius E, Appelkvist P, Strömberg K, Höglund K. Characterization of the effect of a novel γ-secretase modulator on Aβ: a clinically translatable model. Curr Pharm Des. 2014;20:2484–90.CrossRefPubMed

21.

Portelius E, Van Broeck B, Andreasson U, Gustavsson MK, Mercken M, Zetterberg H, et al. Acute effect on the Abeta isoform pattern in CSF in response to gamma-secretase modulator and inhibitor treatment in dogs. J Alzheimers Dis. 2010;21:1005–12. doi:10.3233/JAD-2010-100573.PubMed

22.

Crump CJ, Johnson DS, Li YM. Development and mechanism of gamma-secretase modulators for Alzheimer’s disease. Biochemistry. 2013;52:3197–216. doi:10.1021/bi400377p.CrossRefPubMed

23.

Jacobsen KT, Iverfeldt K. Amyloid precursor protein and its homologues: a family of proteolysis-dependent receptors. Cell Mol Life Sci. 2009;66:2299–318. doi:10.1007/s00018-009-0020-8.CrossRefPubMed

24.

Minogue AM, Stubbs AK, Frigerio CS, Boland B, Fadeeva JV, Tang J, et al. gamma-secretase processing of APLP1 leads to the production of a p3-like peptide that does not aggregate and is not toxic to neurons. Brain Res. 2009;1262:89–99. doi:10.1016/j.brainres.2009.01.008.CrossRefPubMed

25.

Baumkotter F, Wagner K, Eggert S, Wild K, Kins S. Structural aspects and physiological consequences of APP/APLP trans-dimerization. Exp Brain Res. 2012;217:389–95. doi:10.1007/s00221-011-2878-6.PubMedCentralCrossRefPubMed

26.

Eggert S, Paliga K, Soba P, Evin G, Masters CL, Weidemann A, et al. The proteolytic processing of the amyloid precursor protein gene family members APLP-1 and APLP-2 involves alpha-, beta-, gamma-, and epsilon-like cleavages: modulation of APLP-1 processing by n-glycosylation. J Biol Chem. 2004;279:18146–56. doi:10.1074/jbc.M311601200.CrossRefPubMed

27.

Haass C, Schlossmacher MG, Hung AY, Vigo-Pelfrey C, Mellon A, Ostaszewski BL, et al. Amyloid beta-peptide is produced by cultured cells during normal metabolism. Nature. 1992;359:322–5. doi:10.1038/359322a0.CrossRefPubMed

28.

Yanagida K, Okochi M, Tagami S, Nakayama T, Kodama TS, Nishitomi K, et al. The 28-amino acid form of an APLP1-derived Abeta-like peptide is a surrogate marker for Abeta42 production in the central nervous system. EMBO Mol Med. 2009;1:223–35. doi:10.1002/emmm.200900026.PubMedCentralCrossRefPubMed

29.

Naruse S, Thinakaran G, Luo JJ, Kusiak JW, Tomita T, Iwatsubo T, et al. Effects of PS1 deficiency on membrane protein trafficking in neurons. Neuron. 1998;21:1213–21.CrossRefPubMed

30.

Walsh DM, Fadeeva JV, LaVoie MJ, Paliga K, Eggert S, Kimberly WT, et al. gamma-Secretase cleavage and binding to FE65 regulate the nuclear translocation of the intracellular C-terminal domain (ICD) of the APP family of proteins. Biochemistry. 2003;42:6664–73. doi:10.1021/bi027375c.CrossRefPubMed

31.

Li Q, Sudhof TC. Cleavage of amyloid-beta precursor protein and amyloid-beta precursor-like protein by BACE 1. J Biol Chem. 2004;279:10542–50. doi:10.1074/jbc.M310001200.CrossRefPubMed

32.

Tagami S, Okochi M, Yanagida K, Kodama T, Arai T, Kuwano R, et al. Relative ratio and level of amyloid-beta 42 surrogate in cerebrospinal fluid of familial Alzheimer disease patients with presenilin 1 mutations. Neurodegener Dis. 2014;13:166–70. doi:10.1159/000355258.CrossRefPubMed

33.

Kimura T, Kawano K, Doi E, Kitazawa N, Shin K, Miyagawa T, et al. inventors; Cinnamide compound patent PCT Int. Appl. WO2005115990(A1). 2005.

34.

Wilsson-Rahmberg M, Olovson SG, Forshult E. Method for long-term cerebrospinal fluid collection in the conscious dog. J Invest Surg. 1998;11:207–14.CrossRefPubMed

35.

Holtta M, Zetterberg H, Mirgorodskaya E, Mattsson N, Blennow K, Gobom J. Peptidome analysis of cerebrospinal fluid by LC-MALDI MS. PLoS One. 2012;7:e42555. doi:10.1371/journal.pone.0042555.PubMedCentralCrossRefPubMed

36.

Portelius E, Holtta M, Soininen H, Bjerke M, Zetterberg H, Westerlund A, et al. Altered cerebrospinal fluid levels of amyloid beta and amyloid precursor-like protein 1 peptides in Down’s syndrome. Neuromolecular Med. 2014;16:510–6. doi:10.1007/s12017-014-8302-1.CrossRefPubMed

37.

Wanngren J, Ottervald J, Parpal S, Portelius E, Stromberg K, Borgegard T, et al. Second generation gamma-secretase modulators exhibit different modulation of Notch beta and Abeta production. J Biol Chem. 2012;287:32640–50. doi:10.1074/jbc.M112.376541.PubMedCentralCrossRefPubMed

38.

Kukar TL, Ladd TB, Bann MA, Fraering PC, Narlawar R, Maharvi GM, et al. Substrate-targeting gamma-secretase modulators. Nature. 2008;453:925–9. doi:10.1038/nature07055.PubMedCentralCrossRefPubMed

39.

Ebke A, Luebbers T, Fukumori A, Shirotani K, Haass C, Baumann K, et al. Novel γ-secretase enzyme modulators directly target presenilin protein. J Biol Chem. 2011;286:37181–6. doi:10.1074/jbc.C111.276972.PubMedCentralCrossRefPubMed

40.

Pozdnyakov N, Murrey HE, Crump CJ, Pettersson M, Ballard TE, Am Ende CW, et al. Gamma-secretase modulator (GSM) photoaffinity probes reveal distinct allosteric binding sites on presenilin. J Biol Chem. 2013;288:9710–20. doi:10.1074/jbc.M112.398602.PubMedCentralCrossRefPubMed

Title: APLP1 as a cerebrospinal fluid biomarker for γ-secretase modulator treatment
Authors: Simon Sjödin
Kerstin K. A. Andersson
Marc Mercken
Henrik Zetterberg
Herman Borghys
Kaj Blennow
Erik Portelius
Publication date: 01-12-2015
Publisher: BioMed Central
Published in: Alzheimer's Research & Therapy / Issue 1/2015
Electronic ISSN: 1758-9193
DOI: https://doi.org/10.1186/s13195-015-0160-z

At a glance: The STEP trials

Springer Medicine

APLP1 as a cerebrospinal fluid biomarker for γ-secretase modulator treatment

Abstract

Introduction

Methods

Results

Conclusion

At a glance: The STEP trials

Springer Medicine

Abstract

Introduction

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2015

Brain metabolism and cerebrospinal fluid biomarkers profile of non-amnestic mild cognitive impairment in comparison to amnestic mild cognitive impairment and normal older subjects

Association of cognitive function with glucose tolerance and trajectories of glucose tolerance over 12 years in the AusDiab study

Long-term treatment with active Aβ immunotherapy with CAD106 in mild Alzheimer’s disease

Microstate connectivity alterations in patients with early Alzheimer’s disease

Central delivery of iodine-125–labeled cetuximab, etanercept and anakinra after perispinal injection in rats: possible implications for treating Alzheimer’s disease

A highly sensitive novel immunoassay specifically detects low levels of soluble Aβ oligomers in human cerebrospinal fluid