Advertisement

Pharmaceutical Research

, 35:49 | Cite as

Dihydroceramide Desaturase 1 Inhibitors Reduce Amyloid-β Levels in Primary Neurons from an Alzheimer’s Disease Transgenic Model

  • Lara Ordóñez-Gutiérrez
  • Irene Benito-Cuesta
  • José Luis Abad
  • Josefina Casas
  • Gemma Fábrias
  • Francisco Wandosell
Research Paper Theme: Drug Discovery, Development and Delivery in Alzheimer’s Disease
Part of the following topical collections:
  1. Drug Discovery, Development and Delivery in Alzheimer's Disease

ABSTRACT

Purpose

The induction of autophagy has recently been explored as a promising therapeutic strategy to combat Alzheimer’s disease. Among many other factors, there is evidence that ceramides/dihydroceramides act as mediators of autophagy, although the exact mechanisms underlying such effects are poorly understood. Here, we describe how two dihydroceramide desaturase inhibitors (XM461 and XM462) trigger autophagy and reduce amyloid secretion by neurons.

Methods

Neurons isolated from wild-type and APP/PS1 transgenic mice were exposed to the two dihydroceramide desaturase inhibitors to assess their effect on these cell’s protein and lipid profiles.

Results

Both dihydroceramide desaturase inhibitors increased the autophagic vesicles in wild-type neurons, reflected as an increase in LC3-II, and this was correlated with the accumulation of dihydroceramides and dihydrosphingomyelins. Exposing APP/PS1 transgenic neurons to these inhibitors also produced a 50% reduction in amyloid secretion and/or production. The lipidomic defects triggered by these dihydroceramide desaturase inhibitors were correlated with a loss of S6K activity, witnessed by the changes in S6 phosphorylation, which strongly suggested a reduction of mTORC1 activity.

Conclusions

The data obtained strongly suggest that dihydroceramide desaturase 1 activity may modulate autophagy and mTORC1 activity in neurons, inhibiting amyloid secretion and S6K activity. As such, it is tantalizing to propose that dihydroceramide desaturase 1 may be an important therapeutic target to combat amyloidosis.

KEY WORDS

alzheimer’s disease amyloid-β APP/PS1 autophagy dihydroceramide desaturase 1 

ABBREVIATIONS

Amyloid β peptide

AD

Alzheimer’s disease

APP

Amyloid precursor protein

BafA1

Bafilomycin A

Cer

Ceramides

Des1

Dihydroceramide desaturase 1

Des2

Dihydroceramide desaturase 2

dhCer

Dihydroceramides

FAD

Familial alzheimer’s disease

LC3

Microtubule-associated protein 1A/1B–light chain 3

MTT

[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]

p62

Nucleoporin p62

PCR

Polymerase chain reaction

PS1

Presenilin 1

RV

Resveratrol

SLs

Sphingolipids

TGN

Trans golgi network

Notes

ACKNOWLEDGMENTS AND DISCLOSURES

This work was supported by a grant from the Centro de Investigacion Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED; an initiative of the ISCIII). In addition, work in FW’s lab was supported by grants from the “Plan Nacional”, “Dirección General de Ciencia y Tecnología - DGCYT SAF2012-39148-C03-01; and Proyectos I+D+i Retos 2015 SAF2015-70368-R, and an Institutional grant from the” Fundación Areces”.

Supplementary material

11095_2017_2312_Fig8_ESM.gif (104 kb)
Figure S1

(GIF 104 kb)

11095_2017_2312_MOESM1_ESM.eps (1.2 mb)
High resolution image (EPS 1260 kb)

References

  1. 1.
    Selkoe DJ. Alzheimer's disease results from the cerebral accumulation and cytotoxicity of amyloid beta-protein. J Alzheimers Dis. 2001;3(1):75–80.CrossRefPubMedGoogle Scholar
  2. 2.
    Hardy J, Selkoe DJ. The amyloid hypothesis of Alzheimer's disease: progress and problems on the road to therapeutics. Science. 2002;297(5580):353–6.CrossRefPubMedGoogle Scholar
  3. 3.
    Selkoe DJ. Alzheimer's disease: genes, proteins, and therapy. Physiol Rev. 2001;81(2):741–66.CrossRefPubMedGoogle Scholar
  4. 4.
    Sweeney P, Park H, Baumann M, Dunlop J, Frydman J, Kopito R, et al. Protein misfolding in neurodegenerative diseases: implications and strategies. Trans Neurodegener. 2017;6:6.CrossRefGoogle Scholar
  5. 5.
    Ugalde CL, Finkelstein DI, Lawson VA, Hill AF. Pathogenic mechanisms of prion protein, amyloid-beta and alpha-synuclein misfolding: the prion concept and neurotoxicity of protein oligomers. J Neurochem. 2016;139(2):162–80.CrossRefPubMedGoogle Scholar
  6. 6.
    Wileman T. Autophagy as a defence against intracellular pathogens. Essays Biochem. 2013;55:153–63.CrossRefPubMedGoogle Scholar
  7. 7.
    Shibutani ST, Yoshimori T. Autophagosome formation in response to intracellular bacterial invasion. Cell Microbiol. 2014;16(11):1619–26.CrossRefPubMedGoogle Scholar
  8. 8.
    Ryter SW, Cloonan SM, Choi AM. Autophagy: a critical regulator of cellular metabolism and homeostasis. Mol Cells. 2013;36(1):7–16.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Papackova Z, Cahova M. Important role of autophagy in regulation of metabolic processes in health, disease and aging. Physiol Res. 2014;63(4):409–20.PubMedGoogle Scholar
  10. 10.
    Zare-Shahabadi A, Masliah E, Johnson GV, Rezaei N. Autophagy in Alzheimer's disease. Rev Neurosci. 2015;26(4):385–95.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Son JH, Shim JH, Kim KH, Ha JY, Han JY. Neuronal autophagy and neurodegenerative diseases. Exp Mol Med. 2012;44(2):89–98.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Nixon RA, Wegiel J, Kumar A, Yu WH, Peterhoff C, Cataldo A, et al. Extensive involvement of autophagy in Alzheimer disease: an immuno-electron microscopy study. J Neuropathol Exp Neurol. 2005;64(2):113–22.CrossRefPubMedGoogle Scholar
  13. 13.
    Spilman P, Podlutskaya N, Hart MJ, Debnath J, Gorostiza O, Bredesen D, et al. Inhibition of mTOR by rapamycin abolishes cognitive deficits and reduces amyloid-beta levels in a mouse model of Alzheimer's disease. PLoS One. 2010;5(4):e9979.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Caccamo A, Majumder S, Richardson A, Strong R, Oddo S. Molecular interplay between mammalian target of rapamycin (mTOR), amyloid-beta, and Tau: effects on cognitive impairments. J Biol Chem. 2010;285(17):13107–20.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Tommasino C, Marconi M, Ciarlo L, Matarrese P, Malorni W. Autophagic flux and autophagosome morphogenesis require the participation of sphingolipids. Apoptosis: Int J Program Cell Death. 2015;20(5):645–57.CrossRefGoogle Scholar
  16. 16.
    Young MM, Kester M, Wang HG. Sphingolipids: regulators of crosstalk between apoptosis and autophagy. J Lipid Res. 2013;54(1):5–19.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Maceyka M, Spiegel S. Sphingolipid metabolites in inflammatory disease. Nature. 2014;510(7503):58–67.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Casasampere M, Ordonez YF, Casas J, Fabrias G. Dihydroceramide desaturase inhibitors induce autophagy via dihydroceramide-dependent and independent mechanisms. Biochim Biophys Acta. 2017;1861(2):264–75.CrossRefPubMedGoogle Scholar
  19. 19.
    Siddique MM, Li Y, Wang L, Ching J, Mal M, Ilkayeva O, et al. Ablation of dihydroceramide desaturase 1, a therapeutic target for the treatment of metabolic diseases, simultaneously stimulates anabolic and catabolic signaling. Mol Cell Biol. 2013;33(11):2353–69.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Scarlatti F, Sala G, Somenzi G, Signorelli P, Sacchi N, Ghidoni R. Resveratrol induces growth inhibition and apoptosis in metastatic breast cancer cells via de novo ceramide signaling. FASEB J: Off Publ Fed Am Soc Exp Biol. 2003;17(15):2339–41.CrossRefGoogle Scholar
  21. 21.
    Sala G, Minutolo F, Macchia M, Sacchi N, Ghidoni R. Resveratrol structure and ceramide-associated growth inhibition in prostate cancer cells. Drugs Exp Clin Res. 2003;29(5–6):263–9.PubMedGoogle Scholar
  22. 22.
    Anekonda TS. Resveratrol--a boon for treating Alzheimer's disease? Brain Res Rev. 2006;52(2):316–26.CrossRefPubMedGoogle Scholar
  23. 23.
    Munoz-Olaya JM, Matabosch X, Bedia C, Egido-Gabas M, Casas J, Llebaria A, et al. Synthesis and biological activity of a novel inhibitor of dihydroceramide desaturase. ChemMedChem. 2008;3(6):946–53.CrossRefPubMedGoogle Scholar
  24. 24.
    Jankowsky JL, Slunt HH, Ratovitski T, Jenkins NA, Copeland NG, Borchelt DR. Co-expression of multiple transgenes in mouse CNS: a comparison of strategies. Biomol Eng. 2001;17(6):157–65.CrossRefPubMedGoogle Scholar
  25. 25.
    Hong S, Beja-Glasser VF, Nfonoyim BM, Frouin A, Li S, Ramakrishnan S, et al. Complement and microglia mediate early synapse loss in Alzheimer mouse models. Science. 2016;352(6286):712–6.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Kramer D, Minichiello L. Cell culture of primary cerebellar granule cells. Methods Mol Biol. 2010;633:233–9.CrossRefPubMedGoogle Scholar
  27. 27.
    Moreno-Flores MT, Martin-Aparicio E, Martin-Bermejo MJ, Agudo M, McMahon S, Avila J, et al. Semaphorin 3C preserves survival and induces neuritogenesis of cerebellar granule neurons in culture. J Neurochem. 2003;87(4):879–90.CrossRefPubMedGoogle Scholar
  28. 28.
    Deng H, Mi MT. Resveratrol Attenuates Abeta25-35 Caused Neurotoxicity by Inducing Autophagy Through the TyrRS-PARP1-SIRT1 Signaling Pathway. Neurochem Res. 2016;41(9):2367–79.CrossRefPubMedGoogle Scholar
  29. 29.
    Zhao H, Chen S, Gao K, Zhou Z, Wang C, Shen Z, et al. Resveratrol protects against spinal cord injury by activating autophagy and inhibiting apoptosis mediated by the SIRT1/AMPK signaling pathway. Neuroscience. 2017;348:241–51.CrossRefPubMedGoogle Scholar
  30. 30.
    Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65(1–2):55–63.CrossRefPubMedGoogle Scholar
  31. 31.
    Gasparov VS, Degtiar VG. [Protein determination by binding with the dye Coomassie brilliant blue G-250]. Biokhimiia. 1994;59(6):763–77.PubMedGoogle Scholar
  32. 32.
    Hernandez-Tiedra S, Fabrias G, Davila D, Salanueva IJ, Casas J, Montes LR, et al. Dihydroceramide accumulation mediates cytotoxic autophagy of cancer cells via autolysosome destabilization. Autophagy. 2016;12(11):2213–29.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Holliday MW Jr, Cox SB, Kang MH, Maurer BJ. C22:0- and C24:0-dihydroceramides confer mixed cytotoxicity in T-cell acute lymphoblastic leukemia cell lines. PLoS One. 2013;8(9):e74768.CrossRefPubMedGoogle Scholar
  34. 34.
    Kraveka JM, Li L, Szulc ZM, Bielawski J, Ogretmen B, Hannun YA, et al. Involvement of dihydroceramide desaturase in cell cycle progression in human neuroblastoma cells. J Biol Chem. 2007;282(23):16718–28.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Lavieu G, Scarlatti F, Sala G, Carpentier S, Levade T, Ghidoni R, et al. Sphingolipids in macroautophagy. Methods Mol Biol. 2008;445:159–73.CrossRefPubMedGoogle Scholar
  36. 36.
    Pacheco CD, Lieberman AP. Lipid trafficking defects increase Beclin-1 and activate autophagy in Niemann-Pick type C disease. Autophagy. 2007;3(5):487–9.CrossRefPubMedGoogle Scholar
  37. 37.
    Pacheco CD, Kunkel R, Lieberman AP. Autophagy in Niemann-Pick C disease is dependent upon Beclin-1 and responsive to lipid trafficking defects. Hum Mol Genet. 2007;16(12):1495–503.CrossRefPubMedGoogle Scholar
  38. 38.
    Signorelli P, Munoz-Olaya JM, Gagliostro V, Casas J, Ghidoni R, Fabrias G. Dihydroceramide intracellular increase in response to resveratrol treatment mediates autophagy in gastric cancer cells. Cancer Lett. 2009;282(2):238–43.CrossRefPubMedGoogle Scholar
  39. 39.
    Li Y, Li S, Qin X, Hou W, Dong H, Yao L, et al. The pleiotropic roles of sphingolipid signaling in autophagy. Cell Death Dis. 2014;5:e1245.CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Caccamo A, De Pinto V, Messina A, Branca C, Oddo S. Genetic reduction of mammalian target of rapamycin ameliorates Alzheimer's disease-like cognitive and pathological deficits by restoring hippocampal gene expression signature. J Neurosci: Off J Soc Neurosci. 2014;34(23):7988–98.CrossRefGoogle Scholar
  41. 41.
    Vieira CR, Munoz-Olaya JM, Sot J, Jimenez-Baranda S, Izquierdo-Useros N, Abad JL, et al. Dihydrosphingomyelin impairs HIV-1 infection by rigidifying liquid-ordered membrane domains. Chem Biol. 2010;17(7):766–75.CrossRefPubMedGoogle Scholar
  42. 42.
    Haass C, Kaether C, Thinakaran G, Sisodia S. Trafficking and proteolytic processing of APP. Cold Spring Harb Perspect Med. 2012;2(5):a006270.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Vetrivel KS, Thinakaran G. Membrane rafts in Alzheimer's disease beta-amyloid production. Biochim Biophys Acta. 2010;1801(8):860–7.CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Cai H, Wang Y, McCarthy D, Wen H, Borchelt DR, Price DL, et al. BACE1 is the major beta-secretase for generation of Abeta peptides by neurons. Nat Neurosci. 2001;4(3):233–4.CrossRefPubMedGoogle Scholar
  45. 45.
    Sinha S, Anderson JP, Barbour R, Basi GS, Caccavello R, Davis D, et al. Purification and cloning of amyloid precursor protein beta-secretase from human brain. Nature. 1999;402(6761):537–40.CrossRefPubMedGoogle Scholar
  46. 46.
    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(5440):735–41.CrossRefPubMedGoogle Scholar
  47. 47.
    Nilsson P, Saido TC. Dual roles for autophagy: degradation and secretion of Alzheimer's disease Abeta peptide. Bioessays: News Rev Mol Cell Dev Biol. 2014;36(6):570–8.CrossRefGoogle Scholar
  48. 48.
    Ginsberg SD, Mufson EJ, Counts SE, Wuu J, Alldred MJ, Nixon RA, et al. Regional selectivity of rab5 and rab7 protein upregulation in mild cognitive impairment and Alzheimer's disease. J Alzheimer's Dis: JAD. 2010;22(2):631–9.CrossRefGoogle Scholar
  49. 49.
    Takahashi RH, Milner TA, Li F, Nam EE, Edgar MA, Yamaguchi H, et al. Intraneuronal Alzheimer abeta42 accumulates in multivesicular bodies and is associated with synaptic pathology. Am J Pathol. 2002;161(5):1869–79.CrossRefPubMedPubMedCentralGoogle Scholar
  50. 50.
    Jiang W, Ogretmen B. Autophagy paradox and ceramide. Biochim Biophys Acta. 2014;1841(5):783–92.CrossRefPubMedGoogle Scholar
  51. 51.
    Devlin CM, Lahm T, Hubbard WC, Van Demark M, Wang KC, Wu X, et al. Dihydroceramide-based response to hypoxia. J Biol Chem. 2011;286(44):38069–78.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Centro de Biología Molecular “Severo Ochoa” CSIC-UAMMadridSpain
  2. 2.Centro de Investigación Biomédica en Red en Enfermedades Neurodegenerativas (CIBERNED)MadridSpain
  3. 3.Instituto de Química Avanzada de Cataluña (IQAC-CSIC)BarcelonaSpain

Personalised recommendations