Phosphoinositide 3-Kinases Upregulate System xc− via Eukaryotic Initiation Factor 2α and Activating Transcription Factor 4 – A Pathway Active in Glioblastomas and Epilepsy
Publication: Antioxidants & Redox Signaling
Volume 20, Issue Number 18
Abstract
Aims: Phosphoinositide 3-kinases (PI3Ks) relay growth factor signaling and mediate cytoprotection and cell growth. The cystine/glutamate antiporter system xc− imports cystine while exporting glutamate, thereby promoting glutathione synthesis while increasing extracellular cerebral glutamate. The aim of this study was to analyze the pathway through which growth factor and PI3K signaling induce the cystine/glutamate antiporter system xc− and to demonstrate its biological significance for neuroprotection, cell growth, and epilepsy. Results: PI3Ks induce system xc− through glycogen synthase kinase 3β (GSK-3β) inhibition, general control non-derepressible-2-mediated eukaryotic initiation factor 2α phosphorylation, and the subsequent translational up-regulation of activating transcription factor 4. This pathway is essential for PI3Ks to modulate oxidative stress resistance of nerve cells and insulin-induced growth in fibroblasts. Moreover, the pathway is active in human glioblastoma cells. In addition, it is induced in primary cortical neurons in response to robust neuronal activity and in hippocampi from patients with temporal lobe epilepsy. Innovation: Our findings further extend the concepts of how growth factors and PI3Ks induce neuroprotection and cell growth by adding a new branch to the signaling network downstream of GSK-3β, which, ultimately, leads to the induction of the cystine/glutamate antiporter system xc−. Importantly, the induction of this pathway by neuronal activity and in epileptic hippocampi points to a potential role in epilepsy. Conclusion: PI3K-regulated system xc− activity is not only involved in the stress resistance of neuronal cells and in cell growth by increasing the cysteine supply and glutathione synthesis, but also plays a role in the pathophysiology of tumor- and non-tumor-associated epilepsy by up-regulating extracellular cerebral glutamate. Antioxid. Redox Signal. 20: 2907–2922.
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References
1.
Adams CM. Role of the transcription factor ATF4 in the anabolic actions of insulin and the anti-anabolic actions of glucocorticoids. J Biol Chem 282: 16744–16753, 2007.
2.
Al-Mubarak B, Soriano FX, and Hardingham GE. Synaptic NMDAR activity suppresses FOXO1 expression via a cis-acting FOXO binding site: FOXO1 is a FOXO target gene. Channels (Austin) 3: 233–238, 2009.
3.
Baird TD and Wek RC. Eukaryotic initiation factor 2 phosphorylation and translational control in metabolism. Adv Nutr 3: 307–321, 2012.
4.
Banjac A, Perisic T, Sato H, Seiler A, Bannai S, Weiss N, Kolle P, Tschoep K, Issels RD, Daniel PT, Conrad M, and Bornkamm GW. The cystine/cysteine cycle: a redox cycle regulating susceptibility versus resistance to cell death. Oncogene 27: 1618–1628, 2008.
5.
Berger R, Jennewein C, Marschall V, Karl S, Cristofanon S, Wagner L, Vellanki SH, Hehlgans S, Rodel F, Debatin KM, Ludolph AC, and Fulda S. NF-kappaB is required for Smac mimetic-mediated sensitization of glioblastoma cells for gamma-irradiation-induced apoptosis. Mol Cancer Ther 10: 1867–1875, 2011.
6.
Bi M, Naczki C, Koritzinsky M, Fels D, Blais J, Hu N, Harding H, Novoa I, Varia M, Raleigh J, Scheuner D, Kaufman RJ, Bell J, Ron D, Wouters BG, and Koumenis C. ER stress-regulated translation increases tolerance to extreme hypoxia and promotes tumor growth. EMBO J 24: 3470–3481, 2005.
7.
Blume-Jensen P, Janknecht R, and Hunter T. The kit receptor promotes cell survival via activation of PI 3-kinase and subsequent Akt-mediated phosphorylation of Bad on Ser136. Curr Biol 8: 779–782, 1998.
8.
Bourette RP and Rohrschneider LR. Early events in M-CSF receptor signaling. Growth Factors 17: 155–166, 2000.
9.
Buckingham SC, Campbell SL, Haas BR, Montana V, Robel S, Ogunrinu T, and Sontheimer H. Glutamate release by primary brain tumors induces epileptic activity. Nat Med 17: 1269–1274, 2011.
10.
Cao H, Yu S, Yao Z, Galson DL, Jiang Y, Zhang X, Fan J, Lu B, Guan Y, Luo M, Lai Y, Zhu Y, Kurihara N, Patrene K, Roodman GD, and Xiao G. Activating transcription factor 4 regulates osteoclast differentiation in mice. J Clin Invest 120: 2755–2766, 2010.
11.
Chowdhry S, Zhang Y, McMahon M, Sutherland C, Cuadrado A, and Hayes JD. Nrf2 is controlled by two distinct beta-TrCP recognition motifs in its Neh6 domain, one of which can be modulated by GSK-3 activity. Oncogene 32: 3765–3781, 2013.
12.
Chung WJ, Lyons SA, Nelson GM, Hamza H, Gladson CL, Gillespie GY, and Sontheimer H. Inhibition of cystine uptake disrupts the growth of primary brain tumors. J Neurosci 25: 7101–7110, 2005.
13.
Cross DA, Alessi DR, Cohen P, Andjelkovich M, and Hemmings BA. Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature 378: 785–789, 1995.
14.
Davis JB and Maher P. Protein kinase C activation inhibits glutamate-induced cytotoxicity in a neuronal cell line. Brain Res 652: 169–173, 1994.
15.
De Bundel D, Schallier A, Loyens E, Fernando R, Miyashita H, Van Liefferinge J, Vermoesen K, Bannai S, Sato H, Michotte Y, Smolders I, and Massie A. Loss of system xc- does not induce oxidative stress but decreases extracellular glutamate in hippocampus and influences spatial working memory and limbic seizure susceptibility. J Neurosci 31: 5792–5803, 2011.
16.
Furnari FB, Fenton T, Bachoo RM, Mukasa A, Stommel JM, Stegh A, Hahn WC, Ligon KL, Louis DN, Brennan C, Chin L, DePinho RA, and Cavenee WK. Malignant astrocytic glioma: genetics, biology, and paths to treatment. Genes Dev 21: 2683–2710, 2007.
17.
Gharbi SI, Zvelebil MJ, Shuttleworth SJ, Hancox T, Saghir N, Timms JF, and Waterfield MD. Exploring the specificity of the PI3K family inhibitor LY294002. Biochem J 404: 15–21, 2007.
18.
Gorter JA, Zurolo E, Iyer A, Fluiter K, van Vliet EA, Baayen JC, and Aronica E. Induction of sodium channel Na(x) (SCN7A) expression in rat and human hippocampus in temporal lobe epilepsy. Epilepsia 51: 1791–1800, 2010.
19.
Grimes CA and Jope RS. The multifaceted roles of glycogen synthase kinase 3beta in cellular signaling. Prog Neurobiol 65: 391–426, 2001.
20.
Gupta K, Patani R, Baxter P, Serio A, Story D, Tsujita T, Hayes JD, Pedersen RA, Hardingham GE, and Chandran S. Human embryonic stem cell derived astrocytes mediate non-cell-autonomous neuroprotection through endogenous and drug-induced mechanisms. Cell Death Differ 19: 779–787, 2012.
21.
Hafsi S, Pezzino FM, Candido S, Ligresti G, Spandidos DA, Soua Z, McCubrey JA, Travali S, and Libra M. Gene alterations in the PI3K/PTEN/AKT pathway as a mechanism of drug-resistance (review). Int J Oncol 40: 639–644, 2012.
22.
Harding HP, Novoa I, Zhang Y, Zeng H, Wek R, Schapira M, and Ron D. Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol Cell 6: 1099–1108, 2000.
23.
Harding HP, Zhang Y, Zeng H, Novoa I, Lu PD, Calfon M, Sadri N, Yun C, Popko B, Paules R, Stojdl DF, Bell JC, Hettmann T, Leiden JM, and Ron D. An integrated stress response regulates amino acid metabolism and resistance to oxidative stress. Mol Cell 11: 619–633, 2003.
24.
Hardingham GE, Arnold FJ, and Bading H. Nuclear calcium signaling controls CREB-mediated gene expression triggered by synaptic activity. Nat Neurosci 4: 261–267, 2001.
25.
He CH, Gong P, Hu B, Stewart D, Choi ME, Choi AM, and Alam J. Identification of activating transcription factor 4 (ATF4) as an Nrf2-interacting protein. Implication for heme oxygenase-1 gene regulation. J Biol Chem 276: 20858–20865, 2001.
26.
Hui L, Pei DS, Zhang QG, Guan QH, and Zhang GY. The neuroprotection of insulin on ischemic brain injury in rat hippocampus through negative regulation of JNK signaling pathway by PI3K/Akt activation. Brain Res 1052: 1–9, 2005.
27.
Inageda K. Insulin modulates induction of glucose-regulated protein 78 during endoplasmic reticulum stress via augmentation of ATF4 expression in human neuroblastoma cells. FEBS Lett 584: 3649–3654, 2010.
28.
Jackson SP, Schoenwaelder SM, Goncalves I, Nesbitt WS, Yap CL, Wright CE, Kenche V, Anderson KE, Dopheide SM, Yuan Y, Sturgeon SA, Prabaharan H, Thompson PE, Smith GD, Shepherd PR, Daniele N, Kulkarni S, Abbott B, Saylik D, Jones C, Lu L, Giuliano S, Hughan SC, Angus JA, Robertson AD, and Salem HH. PI 3-kinase p110beta: a new target for antithrombotic therapy. Nat Med 11: 507–514, 2005.
29.
Jiang HY and Wek RC. GCN2 phosphorylation of eIF2alpha activates NF-kappaB in response to UV irradiation. Biochem J 385: 371–380, 2005.
30.
Johnson DA, Andrews GK, Xu W, and Johnson JA. Activation of the antioxidant response element in primary cortical neuronal cultures derived from transgenic reporter mice. J Neurochem 81: 1233–1241, 2002.
31.
Kaufman RJ, Davies MV, Pathak VK, and Hershey JW. The phosphorylation state of eucaryotic initiation factor 2 alters translational efficiency of specific mRNAs. Mol Cell Biol 9: 946–958, 1989.
32.
Kilberg MS, Shan J, and Su N. ATF4-dependent transcription mediates signaling of amino acid limitation. Trends Endocrinol Metab 20: 436–443, 2009.
33.
Klein PS and Melton DA. A molecular mechanism for the effect of lithium on development. Proc Natl Acad Sci U S A 93: 8455–8459, 1996.
34.
Knobbe CB and Reifenberger G. Genetic alterations and aberrant expression of genes related to the phosphatidyl-inositol-3′-kinase/protein kinase B (Akt) signal transduction pathway in glioblastomas. Brain Pathol 13: 507–518, 2003.
35.
Lee JM, Hanson JM, Chu WA, and Johnson JA. Phosphatidylinositol 3-kinase, not extracellular signal-regulated kinase, regulates activation of the antioxidant-responsive element in IMR-32 human neuroblastoma cells. J Biol Chem 276: 20011–20016, 2001.
36.
Lewerenz J, Albrecht P, Tien ML, Henke N, Karumbayaram S, Kornblum HI, Wiedau-Pazos M, Schubert D, Maher P, and Methner A. Induction of Nrf2 and xCT are involved in the action of the neuroprotective antibiotic ceftriaxone in vitro. J Neurochem 111: 332–343, 2009.
37.
Lewerenz J, Klein M, and Methner A. Cooperative action of glutamate transporters and cystine/glutamate antiporter system Xc- protects from oxidative glutamate toxicity. J Neurochem 98: 916–925, 2006.
38.
Lewerenz J, Letz J, and Methner A. Activation of stimulatory heterotrimeric G proteins increases glutathione and protects neuronal cells against oxidative stress. J Neurochem 87: 522–531, 2003.
39.
Lewerenz J and Maher P. Basal levels of eIF2alpha phosphorylation determine cellular antioxidant status by regulating ATF4 and xCT expression. J Biol Chem 284: 1106–1115, 2009.
40.
Lewerenz J, Sato H, Albrecht P, Henke N, Noack R, Methner A, and Maher P. Mutation of ATF4 mediates resistance of neuronal cell lines against oxidative stress by inducing xCT expression. Cell Death Differ 19: 847–858, 2012.
41.
Li L, Xu B, Zhu Y, Chen L, and Sokabe M. DHEA prevents Abeta25-Abeta35-impaired survival of newborn neurons in the dentate gyrus through a modulation of PI3K-Akt-mTOR signaling. Neuropharmacology 59: 323–333, 2010.
42.
Liu X, Resch J, Rush T, and Lobner D. Functional upregulation of system xc-by fibroblast growth factor-2. Neuropharmacology 62: 901–906, 2012.
43.
Longo N, Franchi-Gazzola R, Bussolati O, Dall'Asta V, Foa PP, Guidotti GG, and Gazzola GC. Effect of insulin on the activity of amino acid transport systems in cultured human fibroblasts. Biochim Biophys Acta 844: 216–223, 1985.
44.
Lyons SA, Chung WJ, Weaver AK, Ogunrinu T, and Sontheimer H. Autocrine glutamate signaling promotes glioma cell invasion. Cancer Res 67: 9463–9471, 2007.
45.
Maher P, Lewerenz J, Lozano C, and Torres JL. A novel approach to enhancing cellular glutathione levels. J Neurochem 107: 690–700, 2008.
46.
Maher PA. Nuclear translocation of fibroblast growth factor (FGF) receptors in response to FGF-2. J Cell Biol 134: 529–536, 1996.
47.
Malmberg SE and Adams CM. Insulin signaling and the general amino acid control response: two distinct pathways to amino acid synthesis and uptake. J Biol Chem 283: 19229–19234, 2008.
48.
Marone R, Cmiljanovic V, Giese B, and Wymann MP. Targeting phosphoinositide 3-kinase: moving towards therapy. Biochim Biophys Acta 1784: 159–185, 2008.
49.
Massie A, Schallier A, Mertens B, Vermoesen K, Bannai S, Sato H, Smolders I, and Michotte Y. Time-dependent changes in striatal xCT protein expression in hemi-Parkinson rats. Neuroreport 19: 1589–1592, 2008.
50.
McMahon M, Lamont DJ, Beattie KA, and Hayes JD. Keap1 perceives stress via three sensors for the endogenous signaling molecules nitric oxide, zinc, and alkenals. Proc Natl Acad Sci U S A 107: 18838–18843, 2010.
51.
Meister A. Metabolism and function of glutathione. In: Glutathione: Chemical, Biochemical and Medical Aspects, edited by Dolphin D, Poulsen R, and Avramovic O. New York: John Wiley and Sons, 1989, pp. 367–474.
52.
Murphy TH, Miyamoto M, Sastre A, Schnaar RL, and Coyle JT. Glutamate toxicity in a neuronal cell line involves inhibition of cystine transport leading to oxidative stress. Neuron 2: 1547–1558, 1989.
53.
Ogunrinu TA and Sontheimer H. Hypoxia increases the dependence of glioma cells on glutathione. J Biol Chem 285: 37716–37724, 2010.
54.
Papadia S, Soriano FX, Leveille F, Martel MA, Dakin KA, Hansen HH, Kaindl A, Sifringer M, Fowler J, Stefovska V, McKenzie G, Craigon M, Corriveau R, Ghazal P, Horsburgh K, Yankner BA, Wyllie DJ, Ikonomidou C, and Hardingham GE. Synaptic NMDA receptor activity boosts intrinsic antioxidant defenses. Nat Neurosci 11: 476–487, 2008.
55.
Pauly K, Fritz K, Furey A, and Lobner D. Insulin-like growth factor 1 and transforming growth factor-beta stimulate cystine/glutamate exchange activity in dental pulp cells. J Endod 37: 943–947, 2011.
56.
Proud CG. eIF2 and the control of cell physiology. Semin Cell Dev Biol 16: 3–12, 2005.
57.
Proud CG. Regulation of protein synthesis by insulin. Biochem Soc Trans 34: 213–216, 2006.
58.
Quesada A, Lee BY, and Micevych PE. PI3 kinase/Akt activation mediates estrogen and IGF-1 nigral DA neuronal neuroprotection against a unilateral rat model of Parkinson's disease. Dev Neurobiol 68: 632–644, 2008.
59.
Rada P, Rojo AI, Chowdhry S, McMahon M, Hayes JD, and Cuadrado A. SCF/{beta}-TrCP promotes glycogen synthase kinase 3-dependent degradation of the Nrf2 transcription factor in a Keap1-independent manner. Mol Cell Biol 31: 1121–1133, 2011.
60.
Romano PR, Garcia-Barrio MT, Zhang X, Wang Q, Taylor DR, Zhang F, Herring C, Mathews MB, Qin J, and Hinnebusch AG. Autophosphorylation in the activation loop is required for full kinase activity in vivo of human and yeast eukaryotic initiation factor 2alpha kinases PKR and GCN2. Mol Cell Biol 18: 2282–2297, 1998.
61.
Rutkowski DT and Kaufman RJ. All roads lead to ATF4. Dev Cell 4: 442–444, 2003.
62.
Samuels Y and Waldman T. Oncogenic mutations of PIK3CA in human cancers. Curr Top Microbiol Immunol 347: 21–41, 2010.
63.
Sasaki H, Sato H, Kuriyama-Matsumura K, Sato K, Maebara K, Wang H, Tamba M, Itoh K, Yamamoto M, and Bannai S. Electrophile response element-mediated induction of the cystine/glutamate exchange transporter gene expression. J Biol Chem 277: 44765–44771, 2002.
64.
Sato H, Nomura S, Maebara K, Sato K, Tamba M, and Bannai S. Transcriptional control of cystine/glutamate transporter gene by amino acid deprivation. Biochem Biophys Res Commun 325: 109–116, 2004.
65.
Savaskan NE, Heckel A, Hahnen E, Engelhorn T, Doerfler A, Ganslandt O, Nimsky C, Buchfelder M, and Eyupoglu IY. Small interfering RNA-mediated xCT silencing in gliomas inhibits neurodegeneration and alleviates brain edema. Nat Med 14: 629–632, 2008.
66.
Savaskan NE, Seufert S, Hauke J, Trankle C, Eyupoglu IY, and Hahnen E. Dissection of mitogenic and neurodegenerative actions of cystine and glutamate in malignant gliomas. Oncogene 30: 43–53, 2011.
67.
Schafer FQ and Buettner GR. Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. Free Radic Biol Med 30: 1191–1212, 2001.
68.
Scheuner D, Song B, McEwen E, Liu C, Laybutt R, Gillespie P, Saunders T, Bonner-Weir S, and Kaufman RJ. Translational control is required for the unfolded protein response and in vivo glucose homeostasis. Mol Cell 7: 1165–1176, 2001.
69.
Shen J, Wu Y, Xu JY, Zhang J, Sinclair SH, Yanoff M, Xu G, Li W, and Xu GT. ERK- and Akt-dependent neuroprotection by erythropoietin (EPO) against glyoxal-AGEs via modulation of Bcl-xL, Bax, and BAD. Invest Ophthalmol Vis Sci 51: 35–46, 2010.
70.
Sims B, Clarke M, Njah W, Hopkins ES, and Sontheimer H. Erythropoietin-induced neuroprotection requires cystine glutamate exchanger activity. Brain Res 1321: 88–95, 2010.
71.
Sun P, Enslen H, Myung PS, and Maurer RA. Differential activation of CREB by Ca2+/calmodulin-dependent protein kinases type II and type IV involves phosphorylation of a site that negatively regulates activity. Genes Dev 8: 2527–2539, 1994.
72.
Sun X, Yao H, Douglas RM, Gu XQ, Wang J, and Haddad GG. Insulin/PI3K signaling protects dentate neurons from oxygen-glucose deprivation in organotypic slice cultures. J Neurochem 112: 377–388, 2010.
73.
Sweeney G, Garg RR, Ceddia RB, Li D, Ishiki M, Somwar R, Foster LJ, Neilsen PO, Prestwich GD, Rudich A, and Klip A. Intracellular delivery of phosphatidylinositol (3,4,5)-trisphosphate causes incorporation of glucose transporter 4 into the plasma membrane of muscle and fat cells without increasing glucose uptake. J Biol Chem 279: 32233–32242, 2004.
74.
Takeuchi S, Wada K, Toyooka T, Shinomiya N, Shimazaki H, Nakanishi K, Nagatani K, Otani N, Osada H, Uozumi Y, Matsuo H, and Nawashiro H. Increased xCT expression correlates with tumor invasion and outcome in patients with glioblastoma. Neurosurgery 72: 33–41, 2012.
75.
Yuen TI, Morokoff AP, Bjorksten A, D'Abaco G, Paradiso L, Finch S, Wong D, Reid CA, Powell KL, Drummond KJ, Rosenthal MA, Kaye AH, and O'Brien TJ. Glutamate is associated with a higher risk of seizures in patients with gliomas. Neurology 79: 883–889, 2012.
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Published In
Antioxidants & Redox Signaling
Volume 20 • Issue Number 18 • June 20, 2014
Pages: 2907 - 2922
PubMed: 24219064
Copyright
Copyright 2014, Mary Ann Liebert, Inc.
History
Published in print: June 20, 2014
Published online: 28 May 2014
Published ahead of print: 6 February 2014
Published ahead of production: 12 November 2013
Accepted: 12 November 2013
Revision received: 25 October 2013
Received: 30 May 2013
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