title: Induction of the unfolded protein response drives enhanced metabolism and chemoresistance in glioma cells.
abstract: The unfolded protein response (UPR) is an endoplasmic reticulum (ER)-based cytoprotective mechanism acting to prevent pathologies accompanying protein aggregation. It is frequently active in tumors, but relatively unstudied in gliomas. We hypothesized that UPR stress effects on glioma cells might protect tumors from additional exogenous stress (ie, chemotherapeutics), postulating that protection was concurrent with altered tumor cell metabolism. Using human brain tumor cell lines, xenograft tumors, human samples and gene expression databases, we determined molecular features of glioma cell UPR induction/activation, and here report a detailed analysis of UPR transcriptional/translational/metabolic responses. Immunohistochemistry, Western and Northern blots identified elevated levels of UPR transcription factors and downstream ER chaperone targets in gliomas. Microarray profiling revealed distinct regulation of stress responses between xenograft tumors and parent cell lines, with gene ontology and network analyses linking gene expression to cell survival and metabolic processes. Human glioma samples were examined for levels of the ER chaperone GRP94 by immunohistochemistry and for other UPR components by Western blotting. Gene and protein expression data from patient gliomas correlated poor patient prognoses with increased expression of ER chaperones, UPR target genes, and metabolic enzymes (glycolysis and lipogenesis). NMR-based metabolomic studies revealed increased metabolic outputs in glucose uptake with elevated glycolytic activity as well as increased phospholipid turnover. Elevated levels of amino acids, antioxidants, and cholesterol were also evident upon UPR stress; in particular, recurrent tumors had overall higher lipid outputs and elevated specific UPR arms. Clonogenicity studies following temozolomide treatment of stressed or unstressed cells demonstrated UPR-induced chemoresistance. Our data characterize the UPR in glioma cells and human tumors, and link the UPR to chemoresistance possibly via enhanced metabolism. Given the role of the UPR in the balance between cell survival and apoptosis, targeting the UPR and/or controlling metabolic activity may prove beneficial for malignant glioma therapeutics.
authors.affiliation: Department of Neurosurgery, Anschutz Medical Center, University of Colorado Denver, Aurora, Colorado, United States of America.
authors.forename: Laura M
authors.initials: LM
authors.lastname: Epple
authors.forename: Rebecca D
authors.initials: RD
authors.lastname: Dodd
authors.forename: Andrea L
authors.initials: AL
authors.lastname: Merz
authors.forename: Anjelika M
authors.initials: AM
authors.lastname: Dechkovskaia
authors.forename: Matthew
authors.initials: M
authors.lastname: Herring
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Winston
authors.forename: Alex M
authors.initials: AM
authors.lastname: Lencioni
authors.forename: Rae L
authors.initials: RL
authors.lastname: Russell
authors.forename: Helen
authors.initials: H
authors.lastname: Madsen
authors.forename: Meheret
authors.initials: M
authors.lastname: Nega
authors.forename: Nathaniel L
authors.initials: NL
authors.lastname: Dusto
authors.forename: Jason
authors.initials: J
authors.lastname: White
authors.forename: Darell D
authors.initials: DD
authors.lastname: Bigner
authors.forename: Christopher V
authors.initials: CV
authors.lastname: Nicchitta
authors.forename: Natalie J
authors.initials: NJ
authors.lastname: Serkova
authors.forename: Michael W
authors.initials: MW
authors.lastname: Graner
mesh_terms: D000818:Animals; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology; D045744:Cell Line, Tumor; D003606:Dacarbazine / Q000031:analogs & derivatives / Q000494:pharmacology; D004195:Disease Models, Animal; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D059865:Endoplasmic Reticulum Stress / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D053263:Gene Regulatory Networks; D005910:Glioma / Q000235:genetics / Q000378:metabolism* / Q000401:mortality / Q000473:pathology; D006801:Humans; D050155:Lipogenesis; D051379:Mice; D018832:Molecular Chaperones / Q000378:metabolism; D060787:Neoplasm Grading; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D000077204:Temozolomide; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D056811:Unfolded Protein Response / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
chemical_list: D018906:Antineoplastic Agents, Alkylating; D018832:Molecular Chaperones; D012333:RNA, Messenger; D014157:Transcription Factors; D003606:Dacarbazine; D000077204:Temozolomide
references.citation: J Biol Chem. 2004 May 7;279(19):20108-17
references.pmid: 14978030
references.citation: J Neurovirol. 1998 Apr;4(2):148-58
references.pmid: 9584952
references.citation: Mol Cell Biochem. 2005 Dec;280(1-2):1-8
references.pmid: 16511951
references.citation: Exp Cell Res. 2005 Jan 15;302(2):270-80
references.pmid: 15561107
references.citation: Stem Cells. 2011 Dec;29(12):1942-51
references.pmid: 21957016
references.citation: Neoplasia. 1999 Aug;1(3):208-19
references.pmid: 10935475
references.citation: J Biol Chem. 2002 Jun 28;277(26):23314-20
references.pmid: 11964406
references.citation: Oncogene. 2003 May 8;22(18):2823-35
references.pmid: 12743605
references.citation: Microsc Res Tech. 1998 Nov 1;43(3):250-7
references.pmid: 9840803
references.citation: Cancer Res. 2009 Dec 1;69(23):9105-11
references.pmid: 19920201
references.citation: J Biol Chem. 2004 Apr 23;279(17):16954-62
references.pmid: 14960588
references.citation: NMR Biomed. 2003 Oct-Nov;16(6-7):370-99
references.pmid: 14679501
references.citation: Neurosurg Focus. 2006 Apr 15;20(4):E7
references.pmid: 16709038
references.citation: Br J Haematol. 2011 Jun;153(6):741-52
references.pmid: 21517817
references.citation: J Neurosurg. 2007 Mar;106(3):417-27
references.pmid: 17367064
references.citation: Cell Tissue Res. 1999 May;296(2):221-7
references.pmid: 10382266
references.citation: Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7727-31
references.pmid: 8052651
references.citation: AJNR Am J Neuroradiol. 1995 May;16(5):1013-7
references.pmid: 7639121
references.citation: Cancer Res. 1996 Nov 1;56(21):5079-86
references.pmid: 8895767
references.citation: J Neurosci Res. 2002 Jul 15;69(2):197-206
references.pmid: 12111801
references.citation: J Neuroimmunol. 2007 Jan;182(1-2):63-72
references.pmid: 17137636
references.citation: Cancer Cell. 2006 May;9(5):391-403
references.pmid: 16697959
references.citation: EMBO Rep. 2006 Feb;7(2):180-5
references.pmid: 16322758
references.citation: Curr Opin Neurol. 2012 Dec;25(6):774-9
references.pmid: 23007009
references.citation: J Cell Physiol. 2007 Dec;213(3):721-9
references.pmid: 17520696
references.citation: Cancer Res. 2012 Jan 15;72(2):491-502
references.pmid: 22135092
references.citation: Cancer Treat Rev. 2013 Apr;39(2):171-9
references.pmid: 23063837
references.citation: Int J Biochem Cell Biol. 2006 Mar;38(3):317-32
references.pmid: 16290097
references.citation: PLoS One. 2008;3(11):e3655
references.pmid: 18985161
references.citation: Int J Radiat Oncol Biol Phys. 1989 Jun;16(6):1355-65
references.pmid: 2656598
references.citation: Cancer Res. 2009 May 15;69(10):4252-9
references.pmid: 19435898
references.citation: Biochem J. 2005 Dec 1;392(Pt 2):313-24
references.pmid: 16086669
references.citation: J Virol. 2011 Jan;85(2):705-14
references.pmid: 21084482
references.citation: J Biol Chem. 2011 Aug 19;286(33):29408-29416
references.pmid: 21697087
references.citation: Annu Rev Genet. 2012;46:165-83
references.pmid: 22934644
references.citation: Growth Factors. 2011 Oct;29(5):187-95
references.pmid: 21831009
references.citation: Cancer Res. 2012 Jan 1;72(1):304-14
references.pmid: 22084398
references.citation: J Neuropathol Exp Neurol. 2005 Jun;64(6):479-89
references.pmid: 15977639
references.citation: J Biochem. 1997 Sep;122(3):622-6
references.pmid: 9348093
references.citation: PLoS One. 2009 Jun 17;4(6):e5947
references.pmid: 19536297
references.citation: Curr Pharm Des. 2011;17(3):284-92
references.pmid: 21348829
references.citation: Cancer Chemother Pharmacol. 1994;34(2):171-4
references.pmid: 8194169
references.citation: FEBS J. 2009 May;276(10):2752-61
references.pmid: 19389115
references.citation: Biochim Biophys Acta. 2013 Aug;1832(8):1117-28
references.pmid: 23517917
references.citation: Biochim Biophys Acta. 2001 Jun 6;1512(2):335-44
references.pmid: 11406111
references.citation: Cancer Res. 2007 Nov 15;67(22):10631-4
references.pmid: 18006802
references.citation: Cancer Res. 2010 Jun 1;70(11):4499-508
references.pmid: 20460504
references.citation: Nat Neurosci. 2006 Mar;9(3):331-9
references.pmid: 16491078
references.citation: Mol Cell Biol. 2010 Nov;30(21):5033-42
references.pmid: 20733002
references.citation: World J Hepatol. 2010 Jun 27;2(6):203-7
references.pmid: 21160998
references.citation: Ai Zheng. 2006 Mar;25(3):326-9
references.pmid: 16536987
references.citation: Br J Pharmacol. 2009 Sep;158(2):588-600
references.pmid: 19663881
references.citation: Biomed Pharmacother. 2013 Oct;67(8):731-6
references.pmid: 23602052
references.citation: Nat Med. 2004 Jan;10(1):98-102
references.pmid: 14702639
references.citation: Nutrition. 2010 Nov-Dec;26(11-12):1146-50
references.pmid: 20080028
references.citation: Oncogene. 2008 Mar 6;27(11):1618-28
references.pmid: 17828297
references.citation: J Neurooncol. 2003 May;62(3):329-38
references.pmid: 12777086
references.citation: Int J Radiat Oncol Biol Phys. 2004 Mar 1;58(3):927-31
references.pmid: 14967452
references.citation: Annu Rev Biochem. 2005;74:739-89
references.pmid: 15952902
references.citation: Kidney Int. 2005 Mar;67(3):1142-51
references.pmid: 15698456
references.citation: Biochem Biophys Res Commun. 2001 Jan 12;280(1):407-14
references.pmid: 11162531
references.citation: Cell Mol Life Sci. 2009 Sep;66(17):2835-50
references.pmid: 19468685
references.citation: J Neurosci. 2007 Oct 17;27(42):11214-27
references.pmid: 17942716
references.citation: Cancer Res. 1988 Aug 1;48(15):4189-95
references.pmid: 3390813
references.citation: Mol Cancer. 2009 Feb 10;8:6
references.pmid: 19208232
references.citation: J Biol Chem. 2002 Dec 13;277(50):49047-54
references.pmid: 12374812
references.citation: Cell Cycle. 2010 Jul 15;9(14):2719-25
references.pmid: 20676045
references.citation: J Biol Chem. 2007 Jun 22;282(25):18634-18644
references.pmid: 17475624
references.citation: Science. 2009 May 22;324(5930):1029-33
references.pmid: 19460998
references.citation: Nat Rev Cancer. 2004 Dec;4(12):966-77
references.pmid: 15573118
references.citation: Clin Cancer Res. 2004 Oct 1;10(19):6661-8
references.pmid: 15475456
references.citation: Cell Metab. 2012 Oct 3;16(4):473-86
references.pmid: 23040069
references.citation: J Hepatol. 2008 Mar;48(3):471-8
references.pmid: 18191271
references.citation: Br J Cancer. 2009 May 19;100(10):1523-9
references.pmid: 19401686
references.citation: Cancer Res. 2006 Oct 1;66(19):9349-51
references.pmid: 17018585
references.citation: Curr Cancer Drug Targets. 2009 May;9(3):381-90
references.pmid: 19442057
references.citation: FASEB J. 2009 May;23(5):1541-57
references.pmid: 19109410
references.citation: Antioxid Redox Signal. 2007 Dec;9(12):2277-93
references.pmid: 17979528
references.citation: Breast Cancer Res. 2011 Aug 24;13(4):R82
references.pmid: 21864376
references.citation: J Biol Chem. 2007 Mar 9;282(10):7024-34
references.pmid: 17213183
references.citation: Semin Oncol. 2011 Dec;38 Suppl 4:S21-33
references.pmid: 22078645
references.citation: Semin Oncol. 2006 Aug;33(4):457-65
references.pmid: 16890800
references.citation: Oncogene. 2009 Apr 9;28(14):1653-68
references.pmid: 19234488
references.citation: Pharm Res. 2006 Aug;23(8):1687-95
references.pmid: 16841193
references.citation: Int J Obes (Lond). 2012 Sep;36(9):1248-51
references.pmid: 22124452
references.citation: Sci Signal. 2010 Nov 09;3(147):ra81
references.pmid: 21062993
references.citation: FEBS Lett. 2007 Jan 9;581(1):1-7
references.pmid: 17174306
references.citation: Proc Natl Acad Sci U S A. 2013 Mar 19;110(12):4622-7
references.pmid: 23471998
references.citation: Front Pharmacol. 2013 Feb 11;4:10
references.pmid: 23403503
references.citation: Amino Acids. 2005 Mar;28(2):213-9
references.pmid: 15723240
references.citation: Cell Biochem Biophys. 2003;39(3):223-47
references.pmid: 14716078
references.citation: Cancer Lett. 2006 May 18;236(2):164-74
references.pmid: 15955621
references.citation: J Neuropathol Exp Neurol. 2007 Jan;66(1):1-9
references.pmid: 17204931
references.citation: Physiol Rev. 2006 Oct;86(4):1133-49
references.pmid: 17015486
references.citation: World J Biol Chem. 2011 Sep 26;2(9):193-201
references.pmid: 21949569
references.citation: J Biol Chem. 2010 Nov 26;285(48):37716-24
references.pmid: 20858898
references.citation: Biochem Res Int. 2012;2012:216450
references.pmid: 22110961
references.citation: Cancer Res. 2001 May 1;61(9):3599-603
references.pmid: 11325827
references.citation: Cancer Res. 1997 Sep 15;57(18):4130-40
references.pmid: 9307304
references.citation: Mol Cell Biol. 1994 Jun;14(6):3915-26
references.pmid: 8196632
references.citation: Histochem Cell Biol. 2004 May;121(5):371-6
references.pmid: 15108003
references.citation: Cancer Cell. 2003 Jul;4(1):19-29
references.pmid: 12892710
references.citation: Eur J Gastroenterol Hepatol. 2010 Sep;22(9):1099-105
references.pmid: 20177386
references.citation: Mol Biol Cell. 2005 Dec;16(12):5819-31
references.pmid: 16221886
references.citation: Methods Mol Biol. 2008;419:197-214
references.pmid: 18369985
references.citation: Exp Cell Res. 2005 Jun 10;306(2):343-8
references.pmid: 15925590
references.citation: Endocrinology. 2009 Apr;150(4):1688-96
references.pmid: 19074584
references.citation: Free Radic Biol Med. 2004 Jul 1;37(1):71-85
references.pmid: 15183196
references.citation: Trends Biochem Sci. 2001 Apr;26(4):225-9
references.pmid: 11295554
references.citation: Genes Dev. 1999 May 15;13(10):1211-33
references.pmid: 10346810
references.citation: Mol Cancer Ther. 2005 Oct;4(10):1595-604
references.pmid: 16227410
references.citation: Endocrinology. 2011 Jun;152(6):2247-55
references.pmid: 21447637
references.citation: Anticancer Drugs. 1998 Jun;9(5):449-56
references.pmid: 9660543
references.citation: Cell. 2007 Apr 20;129(2):345-57
references.pmid: 17448993
references.citation: Cancer Res. 2004 Oct 15;64(20):7513-25
references.pmid: 15492278
references.citation: J Neuropathol Exp Neurol. 1999 Aug;58(8):795-802
references.pmid: 10446804
references.citation: EMBO J. 2004 Feb 25;23(4):950-8
references.pmid: 14765107
references.citation: Oncol Rep. 2004 Jun;11(6):1219-23
references.pmid: 15138559
references.citation: Neuro Oncol. 2001 Apr;3(2):73-81
references.pmid: 11296483
references.citation: Cancer Res. 2004 Sep 15;64(18):6716-24
references.pmid: 15374989
references.citation: Proc Natl Acad Sci U S A. 2010 Aug 31;107(35):15553-8
references.pmid: 20702765
references.citation: Biochem Biophys Res Commun. 2002 May 24;293(5):1327-32
references.pmid: 12054658
references.citation: Biosci Biotechnol Biochem. 2013;77(4):729-35
references.pmid: 23563539
references.citation: NMR Biomed. 2001 Aug;14(5):307-17
references.pmid: 11477651
references.citation: Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11514-9
references.pmid: 9326641
references.citation: J Clin Oncol. 2005 Apr 1;23(10):2411-22
references.pmid: 15800333
references.citation: Int J Obes (Lond). 2011 Jun;35(6):762-72
references.pmid: 20938440
references.citation: Nat Rev Cancer. 2008 Nov;8(11):851-64
references.pmid: 18846101
references.citation: Int J Oncol. 2011 Jan;38(1):241-7
references.pmid: 21109946
references.citation: Cancer Res. 2007 Oct 15;67(20):9809-16
references.pmid: 17942911
references.citation: Cell Struct Funct. 2007;32(1):41-50
references.pmid: 17507765
references.citation: Cancer Sci. 2009 Oct;100(10):1870-9
references.pmid: 19659607
references.citation: Cell Growth Differ. 1995 Oct;6(10):1251-9
references.pmid: 8845302
references.citation: J Biol Chem. 2007 Jul 27;282(30):21618-28
references.pmid: 17526500
references.citation: Cancer Res. 2002 Jun 15;62(12):3347-50
references.pmid: 12067971
references.citation: J Biol Chem. 2009 Sep 18;284(38):25569-75
references.pmid: 19617629
references.citation: Cancer Cell. 2006 Mar;9(3):157-73
references.pmid: 16530701
references.citation: J Clin Oncol. 2007 Sep 10;25(26):4127-36
references.pmid: 17827463
references.citation: Biochem Biophys Res Commun. 1999 Feb 5;255(1):59-63
references.pmid: 10082655
references.citation: Mol Cancer Res. 2006 Jul;4(7):423-36
references.pmid: 16849518
references.citation: Differentiation. 2006 Dec;74(9-10):481-7
references.pmid: 17177845
references.citation: Mol Biol Cell. 2006 Jul;17(7):3095-107
references.pmid: 16672378
references.citation: Cancer Treat Rev. 2008 Oct;34(6):558-67
references.pmid: 18501520
references.citation: EMBO J. 2005 Oct 5;24(19):3470-81
references.pmid: 16148948
references.citation: J Biol Chem. 2005 Aug 12;280(32):29060-6
references.pmid: 15980064
references.citation: Biochem Biophys Res Commun. 2008 Jan 4;365(1):35-41
references.pmid: 17980703
references.citation: Mol Cell Biochem. 2008 May;312(1-2):71-80
references.pmid: 18259841
references.citation: Biochem Res Int. 2012;2012:738471
references.pmid: 22110962
references.citation: N Engl J Med. 2005 Mar 10;352(10):987-96
references.pmid: 15758009
references.citation: J Nucl Med. 2009 Aug;50(8):1222-8
references.pmid: 19617323
references.citation: Cancer Res. 2006 Aug 15;66(16):7849-53
references.pmid: 16912156
references.citation: Ann Anat. 2010 Sep 20;192(5):309-13
references.pmid: 20801625