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PMID: 23544129 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Inhibition of PIKfyve by YM-201636 dysregulates autophagy and leads to apoptosis-independent neuronal cell death.

PloS one ·Vol. 8 ·No. 3 ·2013-00-00 ·Pages e60152

Martin S, Harper CB, May LM, Coulson EJ, Meunier FA, Osborne SL

Abstract

The lipid phosphatidylinositol 3,5-bisphosphate (PtdIns(3,5)P 2), synthesised by PIKfyve, regulates a number of intracellular membrane trafficking pathways. Genetic alteration of the PIKfyve complex, leading to even a mild reduction in PtdIns(3,5)P 2, results in marked neurodegeneration via an uncharacterised mechanism. In the present study we have shown that selectively inhibiting PIKfyve activity, using YM-201636, significantly reduces the survival of primary mouse hippocampal neurons in culture. YM-201636 treatment promoted vacuolation of endolysosomal membranes followed by apoptosis-independent cell death. Many vacuoles contained intravacuolar membranes and inclusions reminiscent of autolysosomes. Accordingly, YM-201636 treatment increased the level of the autophagosomal marker protein LC3-II, an effect that was potentiated by inhibition of lysosomal proteases, suggesting that alterations in autophagy could be a contributing factor to neuronal cell death.

MeSH Terms
Aminopyridines/pharmacology Animals Apoptosis/drug effects Autophagy Endocytosis/drug effects Endosomes/drug effects,metabolism Heterocyclic Compounds, 3-Ring/pharmacology Hippocampus/cytology Immunohistochemistry Lysosomes/drug effects,metabolism Mice Mice, Inbred C57BL Neuroendocrine Cells/cytology,drug effects Neurons/cytology,drug effects,metabolism,ultrastructure PC12 Cells Phosphatidylinositol 3-Kinases/metabolism Phosphoinositide-3 Kinase Inhibitors Protein Transport/drug effects Rats Vacuoles/drug effects,metabolism
Chemicals
6-amino-N-(3-(4-(4-morpholinyl)pyrido(3',2'-4,5)furo(3,2-d)pyrimidin-2-yl)phenyl)-3-pyridinecarboxamide Aminopyridines Heterocyclic Compounds, 3-Ring Phosphoinositide-3 Kinase Inhibitors Pikfyve protein, mouse
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Martin Sally
The University of Queensland, Queensland Brain Institute, Brisbane, Queensland, Australia.
Harper Callista B
May Linda M
Coulson Elizabeth J
Meunier Frederic A
Osborne Shona L
References (43)
43 references, click to expand
  1. Functional dissociation between PIKfyve-synthesized PtdIns5P and PtdIns(3,5)P2 by means of the PIKfyve inhibitor YM201636.
    Am J Physiol Cell Physiol. 2012 Aug 15;303(4):C436-46 PMID: 22621786
  2. PIKfyve regulates CaV1.2 degradation and prevents excitotoxic cell death.
    J Cell Biol. 2009 Oct 19;187(2):279-94 PMID: 19841139
  3. Monitoring autophagy in mammalian cultured cells through the dynamics of LC3.
    Methods Enzymol. 2009;452:1-12 PMID: 19200872
  4. Mutation of FIG4 causes neurodegeneration in the pale tremor mouse and patients with CMT4J.
    Nature. 2007 Jul 5;448(7149):68-72 PMID: 17572665
  5. Mammalian cell morphology and endocytic membrane homeostasis require enzymatically active phosphoinositide 5-kinase PIKfyve.
    J Biol Chem. 2001 Jul 13;276(28):26141-7 PMID: 11285266
  6. Endocytosis.
    Physiol Rev. 1997 Jul;77(3):759-803 PMID: 9234965
  7. PI(3,5)P(2) controls membrane trafficking by direct activation of mucolipin Ca(2+) release channels in the endolysosome.
    Nat Commun. 2010 Jul 13;1:38 PMID: 20802798
  8. PIKfyve regulation of endosome-linked pathways.
    Traffic. 2009 Jul;10(7):883-93 PMID: 19582903
  9. Liaisons dangereuses: autophagy, neuronal survival and neurodegeneration.
    Curr Opin Neurobiol. 2008 Oct;18(5):504-15 PMID: 18840524
  10. Loss of Vac14, a regulator of the signaling lipid phosphatidylinositol 3,5-bisphosphate, results in neurodegeneration in mice.
    Proc Natl Acad Sci U S A. 2007 Oct 30;104(44):17518-23 PMID: 17956977
  11. Dynamin inhibition blocks botulinum neurotoxin type A endocytosis in neurons and delays botulism.
    J Biol Chem. 2011 Oct 14;286(41):35966-35976 PMID: 21832053
  12. Regioselective approach to phosphatidylinositol 3,5-bisphosphates: syntheses of the native phospholipid and biotinylated short-chain derivative.
    J Org Chem. 2010 Jun 4;75(11):3541-51 PMID: 20443612
  13. Ca2+-regulated pool of phosphatidylinositol-3-phosphate produced by phosphatidylinositol 3-kinase C2alpha on neurosecretory vesicles.
    Mol Biol Cell. 2008 Dec;19(12):5593-603 PMID: 18843041
  14. PIKfyve controls fluid phase endocytosis but not recycling/degradation of endocytosed receptors or sorting of procathepsin D by regulating multivesicular body morphogenesis.
    Mol Biol Cell. 2003 Nov;14(11):4581-91 PMID: 14551253
  15. PIKfyve negatively regulates exocytosis in neurosecretory cells.
    J Biol Chem. 2008 Feb 1;283(5):2804-13 PMID: 18039667
  16. Impact of 17beta-estradiol on cytokine-mediated apoptotic effects in primary hippocampal and neocortical cell cultures.
    Brain Res. 2006 Oct 20;1116(1):64-74 PMID: 16949056
  17. Localized PtdIns 3,5-P2 synthesis to regulate early endosome dynamics and fusion.
    Am J Physiol Cell Physiol. 2006 Aug;291(2):C393-404 PMID: 16510848
  18. The phosphoinositide kinase PIKfyve is vital in early embryonic development: preimplantation lethality of PIKfyve-/- embryos but normality of PIKfyve+/- mice.
    J Biol Chem. 2011 Apr 15;286(15):13404-13 PMID: 21349843
  19. Fab1 phosphatidylinositol 3-phosphate 5-kinase controls trafficking but not silencing of endocytosed receptors.
    Mol Biol Cell. 2006 Sep;17(9):3989-4001 PMID: 16837550
  20. VAC14 nucleates a protein complex essential for the acute interconversion of PI3P and PI(3,5)P(2) in yeast and mouse.
    EMBO J. 2008 Dec 17;27(24):3221-34 PMID: 19037259
  21. Dynamic control of neuroexocytosis by phosphoinositides in health and disease.
    Prog Lipid Res. 2011 Jan;50(1):52-61 PMID: 20705095
  22. The mammalian phosphatidylinositol 3-phosphate 5-kinase (PIKfyve) regulates endosome-to-TGN retrograde transport.
    J Cell Sci. 2006 Oct 1;119(Pt 19):3944-57 PMID: 16954148
  23. Loss of autophagy in the central nervous system causes neurodegeneration in mice.
    Nature. 2006 Jun 15;441(7095):880-4 PMID: 16625205
  24. Fiji: an open-source platform for biological-image analysis.
    Nat Methods. 2012 Jun 28;9(7):676-82 PMID: 22743772
  25. ESCRTs and Fab1 regulate distinct steps of autophagy.
    Curr Biol. 2007 Oct 23;17(20):1817-25 PMID: 17935992
  26. Membrane traffic exploited by protein toxins.
    Annu Rev Cell Dev Biol. 2002;18:1-24 PMID: 12142266
  27. Core protein machinery for mammalian phosphatidylinositol 3,5-bisphosphate synthesis and turnover that regulates the progression of endosomal transport. Novel Sac phosphatase joins the ArPIKfyve-PIKfyve complex.
    J Biol Chem. 2007 Aug 17;282(33):23878-91 PMID: 17556371
  28. Modulation of synaptic function by VAC14, a protein that regulates the phosphoinositides PI(3,5)P₂ and PI(5)P.
    EMBO J. 2012 Aug 15;31(16):3442-56 PMID: 22842785
  29. Maturation of autophagic vacuoles in Mammalian cells.
    Autophagy. 2005 Apr;1(1):1-10 PMID: 16874026
  30. Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3.
    Autophagy. 2007 Sep-Oct;3(5):452-60 PMID: 17534139
  31. Apoptosis in the nervous system.
    Nature. 2000 Oct 12;407(6805):802-9 PMID: 11048732
  32. Inhibition of the PtdIns(5) kinase PIKfyve disrupts intracellular replication of Salmonella.
    EMBO J. 2010 Apr 21;29(8):1331-47 PMID: 20300065
  33. Autophagy gone awry in neurodegenerative diseases.
    Nat Neurosci. 2010 Jul;13(7):805-11 PMID: 20581817
  34. Apoptosis in neurodegenerative disorders.
    Nat Rev Mol Cell Biol. 2000 Nov;1(2):120-9 PMID: 11253364
  35. Caspases: enemies within.
    Science. 1998 Aug 28;281(5381):1312-6 PMID: 9721091
  36. Regulated localization of Rab18 to lipid droplets: effects of lipolytic stimulation and inhibition of lipid droplet catabolism.
    J Biol Chem. 2005 Dec 23;280(51):42325-35 PMID: 16207721
  37. Deleterious variants of FIG4, a phosphoinositide phosphatase, in patients with ALS.
    Am J Hum Genet. 2009 Jan;84(1):85-8 PMID: 19118816
  38. Defective autophagy in neurons and astrocytes from mice deficient in PI(3,5)P2.
    Hum Mol Genet. 2009 Dec 15;18(24):4868-78 PMID: 19793721
  39. Molecular mechanisms of necroptosis: an ordered cellular explosion.
    Nat Rev Mol Cell Biol. 2010 Oct;11(10):700-14 PMID: 20823910
  40. A selective PIKfyve inhibitor blocks PtdIns(3,5)P(2) production and disrupts endomembrane transport and retroviral budding.
    EMBO Rep. 2008 Feb;9(2):164-70 PMID: 18188180
  41. Beta-amyloid(1-42) induces neuronal death through the p75 neurotrophin receptor.
    J Neurosci. 2008 Apr 9;28(15):3941-6 PMID: 18400893
  42. Neuronal expression of Fig4 is both necessary and sufficient to prevent spongiform neurodegeneration.
    Hum Mol Genet. 2012 Aug 15;21(16):3525-34 PMID: 22581779
  43. The phosphoinositide kinase PIKfyve/Fab1p regulates terminal lysosome maturation in Caenorhabditis elegans.
    Mol Biol Cell. 2006 Jul;17(7):3062-74 PMID: 16801682
Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0060152
Published
2013-00-00
Epub
2013-00-27
Pages
e60152
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC3609765
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pone.0060152 PMC Full Text View journal details
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