Abstract
Cell shrinkage and loss of cell viability by apoptosis have been examined in cultured CD95(Fas/Apo-1)-expressing leukemia-derived CEM and HL-60 cells subjected to acute deprivation of glutamine, a major compatible osmolyte engaged in cell volume control. Glutamine deprivation-mediated cell shrinkage promoted a ligand-independent activation of the CD95-mediated apoptotic pathway. Cell transfection with plasmids expressing FADD-DN or v-Flip viral proteins pointed to a functional clustering of CD95 receptors at the cell surface with activation of the ’extrinsic pathway‘ caspase cascade. Accordingly, cell shrinkage did not induce apoptosis in CD95 receptor-negative lymphoma L1210 cells. Replacement of glutamine with surrogate compatible osmolytes counteracted cell volume decrement and protected the CD95-expressing cells from apoptosis. A glutamine deprivation-dependent cell shrinkage with activation of the CD95-mediated pathway was also observed when asparaginase was added to the medium. Asparagine depletion had no role in this process. The cell-size shrinkage-dependent apoptosis induced by glutamine restriction in CD95-expressing leukemic cells may therefore be of clinical relevance in amidohydrolase enzyme therapies. Cell Death and Differentiation (2001) 8, 1004–1013
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- ALL:
-
acute lymphoblastic leukemia
- CD95L:
-
CD95 ligand
- DISC:
-
death-inducing signaling complex
- FADD:
-
Fas-associated death domain protein
- FADD-DN:
-
dominant negative FADD
- GFP:
-
green fluorescent protein
- OMG:
-
3-o-methyl-D-glucose
- TNFR:
-
tumor necrosis factor receptor
- z-DEVD:
-
Asp-Glu-Val-Asp
- z-IETD:
-
Iso-Glu-Thr-Asp
References
Kovacevic Z, Morris HP . 1972 The role of glutamine in the oxidative metabolism of malignant cells Cancer Res. 32: 326–333
Windmueller HG, Spaeth AE . 1974 Uptake and metabolism of plasma glutamine by the small intestine J. Biol. Chem. 249: 5070–5079
Reitzer LJ, Wice BM, Kennell D . 1979 Evidence that glutamine, not sugar, is the major energy source for cultured HeLa cells J. Biol. Chem. 254: 2669–2676
Zielke HR, Zielke CL, Ozand PT . 1984 Glutamine: a major energy source for cultured mammalian cells Fed. Proc. 43: 121–125
Ardawi MSM . 1988 Glutamine and glucose metabolism in human peripheral lymphocytes Metabolism 37: 99–103
Brand K . 1985 Glutamine and glucose metabolism during thymocyte proliferation Biochem. J. 228: 353–361
Szondy Z, Newsholme EA . 1989 The effect of glutamine concentration on the activity of carbamoyl-phosphate synthase II and on the incorporation of [3H]thymidine into DNA in rat mesenteric lymphocytes stimulated by phytohaemagglutinin Biochem. J. 261: 979–983
Hörig H, Spagnoli GC, Filgueira L, Babst R, Gallati H, Harder F, Juretic A, Heberer M . 1993 Exogenous glutamine requirement is confined to late events of T cell activation J. Cell. Biochem. 53: 343–351
Calder PC, Yaqoob P . 1999 Glutamine and the immune system Amino Acids 17: 227–241
Kitoh T, Kubota M, Takimoto T, Hashimoto H, Shimizu T, Sano H, Akiyama Y, Mikawa H . 1990 Metabolic basis for differential glutamine requirements of human leukemia cell lines J. Cell. Physiol. 143: 150–153
Colquhoun A, Newsholme EA . 1997 Aspects of glutamine metabolism in human tumour cells Biochem. Mol. Biol. Int. 41: 583–596
Gazzola GC, Dall'Asta V, Nucci FA, Rossi PA, Bussolati O, Hoffmann EK, Guidotti GG . 1991 Role of amino acid transport system A in the control of cell volume in cultured human fibroblasts Cell. Physiol. Biochem. 1: 131–142
Dall'Asta V, Rossi PA, Bussolati O, Gazzola GC . 1994 Response of human fibroblasts to hypertonic stress. Cell shrinkage is counteracted by an enhanced active transport of neutral amino acids J. Biol. Chem. 269: 10485–10491
Varoqui H, Zhu H, Yao D, Ming H, Erickson JD . 2000 Cloning and functional identification of a neuronal glutamine transporter J. Biol. Chem. 275: 4049–4054
Guidotti GG, Alfieri R, Urbani S, Petronini PG, Borghetti AF . 1994 Engagement in apoptosis of lymphoma-leukaemia cell lines by glutamine deprivation FASEB J. 8: A807
Petronini PG, Urbani S, Alfieri R, Borghetti AF, Guidotti GG . 1996 Cell susceptibility to apoptosis by glutamine deprivation and rescue: Survival and apoptotic death in cultured lymphoma-leukemia cell lines J. Cell. Physiol. 169: 175–185
Urbani S, Fumarola C, Ponderato N, Alfieri R, Petronini PG, Borghetti AF, Guidotti GG . 1996 c-Myc has a role in glutamine deprivation-induced apoptosis of CEM cells Fundam. Clin. Immunol. 4: 27–30
Hueber AO, Zörnig M, Lyon D, Suda T, Nagata S, Evan G . 1997 Requirement for the CD95 receptor-ligand pathway in c-Myc-induced apoptosis Science 278: 1305–1309
Juin P, Hueber AO, Littlewood T, Evan G . 1999 c-Myc-induced sensitization to apoptosis is mediated through cytochrome c release Gene Dev. 13: 1367–1381
Golstein P . 2000 FasL binds preassembled Fas Science 288: 2328–2329
Krammer PH . 2000 CD95's deadly mission in the immune system Nature 407: 789–795
Kischkel FC, Hellbardt S, Behrmann I, Germer M, Pawlita M, Krammer PH, Peter ME . 1995 Cytotoxicity-dependent APO-1 (Fas/CD95)-associated proteins form a death-inducing signaling complex (DISC) with the receptor EMBO J. 14: 5579–5588
Scaffidi C, Fulda S, Srinivansan A, Friesen C, Li F, Tomaselli KJ, Debatin KM, Krammer PH, Peter ME . 1998 Two CD95 (Apo-1/Fas) signaling pathways EMBO J. 17: 1675–1687
Luo X, Budihardjo I, Zou H, Slaughter C, Wang X . 1998 Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors Cell 94: 481–490
Li H, Zhu H, Xu C, Yuan J . 1998 Cleavage of BID by caspase-8 mediates the mitochondrial damage in the Fas pathway of apoptosis Cell 94: 491–501
Rehemtulla A, Hamilton CA, Chinnaiyan AM, Dixit VM . 1997 Ultraviolet radiation-induced apoptosis is mediated by activation of CD-95 (Fas/Apo-1) J. Biol. Chem. 272: 25783–25786
Aragane Y, Kulms D, Metze D, Wilkes G, Poppelmann B, Luger TA, Schwarz T . 1998 Ultraviolet light induces apoptosis via direct activation of CD95 (Fas/Apo-1) independently of its ligand CD95L J. Cell. Biol. 140: 171–182
Faubion WA, Guicciardi ME, Miyoshi H, Bronk SF, Roberts PJ, Svingen PA, Kaufmann SH, Gores GH . 1999 Toxic bile salts induce rodent hepatocyte apoptosis via direct activation of Fas J. Clin. Invest. 103: 137–145
Micheau O, Solary E, Hammann A, Dimanche-Boitrel MT . 1999 Fas ligand-independent, FADD-mediated activation of the Fas death pathway by anticancer drugs J. Biol. Chem. 274: 7987–7992
Sheikh MS, Antinore MJ, Huang Y, Fornace AJ Jr . 1998 Ultraviolet-irradiation-induced apoptosis is mediated via ligand independent activation of tumor necrosis factor receptor 1 Oncogene 17: 2555–2563
Roberts J, Holcenberg JS, Dolowy WC . 1992 Isolation, crystallization, and properties of Achromobacteraceae glutaminase-asparaginase with antitumor activity J. Biol. Chem. 247: 84–90
Roberts J . 1976 Purification and properties of a highly potent antitumor glutaminase-asparaginase from Pseudomonas 7A J. Biol. Chem. 251: 2119–2123
Boos J, Werber G, Ahlke E, Schulze-Westhoff P, Nowak-Göttl U, Würthwein G, Verspohl EJ, Ritter J, Jürgens H . 1996 Monitoring of asparaginase activity and asparagine levels in children on different asparaginase preparations Eur. J. Cancer 32A: 1544–1550
Muller HJ, Boos J . 1998 Use of L-asparaginase in childhood ALL Crit. Rev. Oncol. Hematol. 28: 97–113
Amylon MD, Shuster J, Pullen J, Berard C, Link MP, Wharam M, Katz J, Yu A, Laver J, Ravindranath Y, Kurtzberg J, Desai S, Camitta B, Murphy SB . 1999 Intensive high-dose asparaginase consolidation improves survival for pediatric patients with T cell acute lymphoblastic leukemia and advanced stage lymphoblastic lymphoma: a Pediatric Oncology Group study Leukemia 13: 335–342
Spiers ASD, Wade HE . 1976 Bacterial glutaminase in treatment of acute leukemia Br. Med. J. 1: 1317–1319
Ollenschläger G, Roth E, Linkesch W, Jansen S, Simmel A, Mödder B . 1988 Asparaginase-induced derangements of glutamine metabolism: the pathogenetic basis for some drug-related side-effects Eur. J. Clin. Invest. 18: 512–516
Kitoh T, Asai S, Akiyama Y, Kubota M, Mikawa H . 1992 The inhibition of lymphocyte blastogenesis by asparaginase: critical role of glutamine in both T and B lymphocye transformation Acta Pediatr. Jpn. 34: 579–583
Mariani SM, Matiba B, Baumler C, Krammer PH . 1995 Regulation of cell surface Apo-1/Fas (CD95) ligand expression by metalloproteases Eur. J. Immunol. 25: 2303–2307
Law RO, Turner DPJ . 1987 Are ninhydrin-positive substances volume-regulatory osmolytes in rat renal papillary cells? J. Physiol. 386: 45–61
Gómez-Angelats M, Bortner CD, Cidlowski JA . 2000 Protein kinase C (PKC) inhibits Fas receptor-induced apoptosis through modulation of the loss of K+ and cell shrinkage J. Biol. Chem. 275: 19609–19619
Guidotti GG, Gazzola GC . 1992 Amino acid transporters: Systematic approach and principles of control. In Mammalian Amino Acid Transport. Mechanisms and Control, Kilberg MS and Häussinger D, eds New York: Plenum Press pp. 3–29
Szabo I, Lepple-Weinhues A, Kaba KN, Zoratti M, Gulbins E, Lang F . 1998 Thyrosine kinase-dependent activation of a chloride channel in CD95-induced apoptosis in T lymphocytes Proc. Natl. Acad. Sci. USA 95: 6169–6174
Maeno E, Ishizaki Y, Kanaseki T, Hazama A, Okada Y . 2000 Normotonic cell shrinkage because of disordered volume regulation is an early prerequisite to apoptosis Proc. Natl. Acad. Sci. USA 97: 9487–9492
Ping Yu S, Choi DW . 2000 Ions, cell volume, and apoptosis Proc. Natl. Acad. Sci. USA 97: 9360–9362
Wajant H, Johannes FJ, Haas E, Siemienski K, Schwenzer R, Schubert G, Weiss T, Grell M, Scheurich P . 1998 Dominant-negative FADD inhibits TNFR60-, Fas/Apo1- and TRAIL-R/Apo2-mediated cell death but not gene induction Curr. Biol. 8: 113–116
Hu S, Vincenz C, Buller M, Dixit VM . 1997 A novel family of viral death effector domain-containing molecules that inhibit both CD-95- and tumor necrosis factor receptor-1-induced apoptosis J. Biol. Chem. 272: 9621–9624
Bertin J, Armstrong RC, Ottilie S, Martin DA, Wang Y, Banks S, Wang GH, Senkevich TG, Alnemri ES, Moss B, Lenardo MJ, Tomaselli KJ, Cohen JI . 1997 Death effector domain-containing herpesvirus and poxvirus proteins inhibit both Fas and TNFR-1-induced apoptosis Proc. Natl. Acad. Sci. USA 94: 1172–1176
Mashburn LT, Wriston JC Jr . 1964 Tumor inhibition effect of L-asparaginase from Escherichia Coli Arch. Biochem. Biophys. 105: 450–452
Wade HE, Elsworth R, Herbert D, Keppie J, Sargeant K . 1968 A new L-asparaginase with antitumour activity? Lancet 2: 776–777
Rosette C, Karin M . 1996 Ultraviolet light and osmotic stress: activation of the JNK cascade through multiple growth factor and cytokine receptors Science 274: 1194–1197
Papoff G, Hausler P, Eramo A, Pagano MG, Di Leve G, Signore A, Ruberti G . 1999 Identification and characterization of a ligand-independent oligomerization domain in the extracellular region of the CD95 death receptor J. Biol. Chem. 274: 38241–38250
Siegel RM, Frederiksen JK, Zacharias DA, Ka-Ming Chan F, Johnson M, Lynch D, Tsien RY, Lenardo MJ . 2000 Fas preassociation required for apoptosis signaling and dominant inhibition by pathogenic mutations Science 288: 2354–2357
Memon SA, Hou J, Moreno MB, Zacharchuk CM . 1998 Cutting edge: apoptosis induced by a chimeric Fas/FLICE receptor: lack of requirement for Fas- or FADD-binding proteins J. Immunol. 160: 2046–2049
Bortner CD, Cidlowski JA . 1996 Absence of volume regulatory mechanisms contributes to the rapid activation of apoptosis in thymocytes Am. J. Physiol. 271: C950–C961
Tsukada N, Kobata T, Aizawa Y, Yagita H, Okumura K . 1999 Graft-versus-leukemia effect and graft-versus-host disease can be differentiated by cytotoxic mechanisms in a murine model of allogeneic bone marrow transplantation Blood 93: 2738–2747
Marsters SA, Sheridan JP, Pitti RM, Brush J, Goddard A, Ashkenazi A . 1998 Identification of a ligand for the death-domain-containing receptor Apo3 Curr. Biol. 8: 525–528
Kischkel FC, Lawrence DA, Chuntharapai A, Schow P, Kim KJ, Ashkenazi A . 2000 Apo2L/TRAIL-dependent recruitment of endogenous FADD and caspase-8 to death receptors 4 and 5 Immunity 12: 611–620
Elledge SJ . 1996 Cell cycle checkpoints: preventing an identity crisis Science 274: 1664–1672
Story MD, Voehringer DW, Stephens LC, Meyn RE . 1993 L-asparaginase kills lymphoma cells by apoptosis Cancer Chemother. Pharmacol. 32: 129–133
Ueno T, Ohtawa K, Mitsui K, Kodera Y, Hiroto M, Matsushima A, Inada Y, Nishimura H . 1997 Cell cycle arrest and apoptosis of leukemia cells induced by L-asparaginase Leukemia 11: 1858–1861
Karawajew L, Wuchter C, Ruppert V, Drexler H, Gruss HJ, Dorken B, Ludwig WD . 1997 Differential CD95 expression and function in T and B lineage acute lymphoblastic leukemia cells Leukemia 11: 1245–1252
Papaconstantinou HT, Hwang KO, Rajaraman S, Hellmich MR, Townsend CM Jr, Ko TC . 1998 Glutamine deprivation induces apoptosis in intestinal epithelial cells Surgery 124: 152–160
Bushman JE, Palmieri D, Whinna HC, Church FC . 2000 Insight into the mechanism of asparaginase-induced depletion of antithrombin III in treatment of childhood acute lymphoblastic leukemia Leuk. Res. 24: 559–565
Korinthenberg R, Ullrich K, Ritter J, Stephani U . 1990 Electrolytes, amino acids and proteins in lumbar CSF during the treatment of acute leukemia in childhood Acta Paediatr. Scand. 79: 335–342
Bossi G, Griffiths GM . 1999 Degranulation plays an essential part in regulating cell surface expression of Fas ligand in T cells and natural killer cells Nature Med. 5: 90–96
Acknowledgements
This work was supported by Ministero dell'Università e della Ricerca Scientifica e Tecnologica, (Murst 1999), Roma, Italy.
Author information
Authors and Affiliations
Corresponding author
Additional information
Edited by T Cotter
Rights and permissions
About this article
Cite this article
Fumarola, C., Zerbini, A. & Guidotti, G. Glutamine deprivation-mediated cell shrinkage induces ligand-independent CD95 receptor signaling and apoptosis. Cell Death Differ 8, 1004–1013 (2001). https://doi.org/10.1038/sj.cdd.4400902
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/sj.cdd.4400902
Keywords
This article is cited by
-
Combined inhibition of histone deacetylase and cytidine deaminase improves epigenetic potency of decitabine in colorectal adenocarcinomas
Clinical Epigenetics (2023)
-
Inhibition of extranodal NK/T-cell lymphoma by Chiauranib through an AIF-dependent pathway and its synergy with L-asparaginase
Cell Death & Disease (2023)
-
Asparaginase pharmacology: challenges still to be faced
Cancer Chemotherapy and Pharmacology (2017)
-
Ammonium ions improve the survival of glutamine-starved hybridoma cells
Cell & Bioscience (2016)
-
Influence of partial and complete glutamine-and glucose deprivation of breast-and cervical tumorigenic cell lines
Cell & Bioscience (2015)
