title: Chemotherapeutic drugs induce ATP release via caspase-gated pannexin-1 channels and a caspase/pannexin-1-independent mechanism.
abstract: Anti-tumor immune responses have been linked to the regulated release of ATP from apoptotic cancer cells to engage P2 purinergic receptor signaling cascades in nearby leukocytes. We used the Jurkat T cell acute lymphocytic leukemia model to characterize the role of pannexin-1 (Panx1) channels in the release of nucleotides during chemotherapeutic drug-induced apoptosis. Diverse pro-apoptotic drugs, including topoisomerase II inhibitors, kinase inhibitors, and proteosome inhibitors, induced functional activation of Panx1 channels via caspase-3-mediated cleavage of the Panx1 autoinhibitory C-terminal domain. The caspase-activated Panx1 channels mediated efflux of ATP, but also ADP and AMP, with the latter two comprising >90% of the released adenine nucleotide pool as cells transitioned from the early to late stages of apoptosis. Chemotherapeutic drugs also activated an alternative caspase- and Panx1-independent pathway for ATP release from Jurkat cells in the presence of benzyloxycarbonyl-VAD, a pan-caspase inhibitor. Comparison of Panx1 levels indicated much higher expression in leukemic T lymphocytes than in normal, untransformed T lymphoblasts. This suggests that signaling roles for Panx1 may be amplified in leukemic leukocytes. Together, these results identify chemotherapy-activated pannexin-1 channels and ATP release as possible mediators of paracrine interaction between dying tumor cells and the effector leukocytes that mediate immunogenic anti-tumor responses.
authors.affiliation: Departments of Pharmacology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106.
authors.forename: Andrea
authors.initials: A
authors.lastname: Boyd-Tressler
authors.affiliation: Department of Anatomy and Cell Biology, University of Western Ontario, London, Ontario N6A-SC1, Canada.
authors.forename: Silvia
authors.initials: S
authors.lastname: Penuela
authors.affiliation: Department of Anatomy and Cell Biology, University of Western Ontario, London, Ontario N6A-SC1, Canada.
authors.forename: Dale W
authors.initials: DW
authors.lastname: Laird
authors.affiliation: Departments of Pharmacology, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106; Departments of Physiology and Biophysics and Case Western Reserve University School of Medicine, Cleveland, Ohio 44106; Case Comprehensive Cancer Center, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106 and. Electronic address: george.dubyak@case.edu.
authors.forename: George R
authors.initials: GR
authors.lastname: Dubyak
mesh_terms: D000244:Adenosine Diphosphate / Q000235:genetics / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism*; D061945:Caspase Inhibitors / Q000494:pharmacology*; D017630:Connexins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism* / Q000473:pathology
chemical_list: D000970:Antineoplastic Agents; D061945:Caspase Inhibitors; D017630:Connexins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; C492764:PANX1 protein, human; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; C505407:CASP3 protein, human; D053148:Caspase 3
keywords: ATP; Apoptosis; Caspase; Pannexin; Purinergic Agonist
references.citation: J Biol Chem. 2010 Apr 2;285(14):10850-61
references.pmid: 20123989
references.citation: J Biol Chem. 2012 Mar 30;287(14):11303-11
references.pmid: 22311983
references.citation: J Auton Nerv Syst. 2000 Jul 3;81(1-3):64-8
references.pmid: 10869702
references.citation: J Immunol. 2011 Jun 1;186(11):6553-61
references.pmid: 21508259
references.citation: J Leukoc Biol. 2009 Mar;85(3):526-38
references.pmid: 19095735
references.citation: FASEB J. 2009 Jun;23(6):1685-93
references.pmid: 19211924
references.citation: Nature. 2010 Oct 14;467(7317):863-7
references.pmid: 20944749
references.citation: Br J Pharmacol. 2007 Sep;152(1):141-50
references.pmid: 17603550
references.citation: Proc Natl Acad Sci U S A. 2012 Jul 10;109(28):E1963-71
references.pmid: 22711817
references.citation: Nat Immunol. 2012 Mar 18;13(4):343-51
references.pmid: 22430787
references.citation: Am J Physiol Cell Physiol. 2012 Nov 1;303(9):C924-35
references.pmid: 22785119
references.citation: J Cell Sci. 2010 Apr 15;123(Pt 8):1363-72
references.pmid: 20332104
references.citation: Science. 2011 Dec 16;334(6062):1573-7
references.pmid: 22174255
references.citation: Proc Natl Acad Sci U S A. 2006 May 16;103(20):7655-9
references.pmid: 16682648
references.citation: Cell Death Differ. 2014 Jan;21(1):79-91
references.pmid: 23852373
references.citation: J Biol Chem. 2004 Mar 19;279(12):11875-81
references.pmid: 14702343
references.citation: Cancer Res. 2007 Feb 15;67(4):1545-54
references.pmid: 17308093
references.citation: Trends Immunol. 2012 May;33(5):231-7
references.pmid: 22487321
references.citation: Pflugers Arch. 2012 Apr;463(4):585-92
references.pmid: 22311122
references.citation: J Biol Chem. 2000 Jan 14;275(2):992-8
references.pmid: 10625637
references.citation: Nat Rev Immunol. 2011 Mar;11(3):201-12
references.pmid: 21331080
references.citation: J Biol Chem. 2012 Aug 17;287(34):29184-93
references.pmid: 22753409
references.citation: Curr Opin Oncol. 2009 Jan;21(1):71-6
references.pmid: 19125021
references.citation: J Cell Sci. 2012 Nov 15;125(Pt 22):5514-23
references.pmid: 22956545
references.citation: Purinergic Signal. 2012 Sep;8(3):359-73
references.pmid: 22528679
references.citation: J Biol Chem. 2010 Mar 19;285(12):9147-60
references.pmid: 20086016
references.citation: Mol Pharmacol. 1999 May;55(5):787-94
references.pmid: 10220556
references.citation: Neuron. 2008 Jul 31;59(2):190-4
references.pmid: 18667147
references.citation: Elife. 2014 Mar 25;3:e02172
references.pmid: 24668173
references.citation: FASEB J. 2012 Jan;26(1):376-86
references.pmid: 21926236
references.citation: Nat Med. 2009 Oct;15(10):1170-8
references.pmid: 19767732
references.citation: Mol Pharmacol. 2003 Oct;64(4):785-95
references.pmid: 14500734
references.citation: FEBS Lett. 2004 Aug 13;572(1-3):65-8
references.pmid: 15304325
references.citation: Proc Natl Acad Sci U S A. 2010 Jan 26;107(4):1547-52
references.pmid: 20080644
references.citation: Biochim Biophys Acta. 2014 Jan;1843(1):150-62
references.pmid: 23466868
references.citation: Cell Microbiol. 2012 Nov;14(11):1697-706
references.pmid: 22882764
references.citation: FEBS Lett. 2002 May 22;519(1-3):153-8
references.pmid: 12023035
references.citation: Proc Natl Acad Sci U S A. 2013 Sep 3;110(36):14711-6
references.pmid: 23964122
references.citation: PLoS One. 2014 Jun 09;9(6):e99596
references.pmid: 24911976
references.citation: J Immunol. 2011 Jun 15;186(12):7144-55
references.pmid: 21593381
references.citation: Proc Natl Acad Sci U S A. 2008 Apr 15;105(15):5683-6
references.pmid: 18375752
references.citation: Nature. 2014 Mar 20;507(7492):329-34
references.pmid: 24646995
references.citation: Am J Physiol Cell Physiol. 2009 Feb;296(2):C250-5
references.pmid: 18945939
references.citation: J Biol Chem. 2012 Mar 9;287(11):8407-16
references.pmid: 22267745
references.citation: Biochim Biophys Acta. 2013 Jan;1828(1):15-22
references.pmid: 22305965
references.citation: J Biol Chem. 2010 Jun 4;285(23):17406-16
references.pmid: 20382737
references.citation: J Biol Chem. 2010 Aug 6;285(32):24420-31
references.pmid: 20516070
references.citation: Biochem J. 2002 Oct 1;367(Pt 1):121-8
references.pmid: 12099890
references.citation: Cell Cycle. 2009 Nov 15;8(22):3723-8
references.pmid: 19855167
references.citation: Channels (Austin). 2014;8(2):103-9
references.pmid: 24419036
references.citation: Biochim Biophys Acta. 2013 Jan;1828(1):35-50
references.pmid: 22796188
references.citation: Immunity. 2013 Apr 18;38(4):729-41
references.pmid: 23562161
references.citation: Cell Death Differ. 2013 Feb;20(2):343-52
references.pmid: 23197294
references.citation: Autophagy. 2012 Mar;8(3):413-5
references.pmid: 22361584
references.citation: EMBO J. 1998 Jan 2;17(1):37-49
references.pmid: 9427739
references.citation: J Cell Sci. 2012 Oct 1;125(Pt 19):4567-75
references.pmid: 22767503
references.citation: Am J Physiol Cell Physiol. 2012 Nov 1;303(9):C913-5
references.pmid: 22932686
references.citation: Nature. 2013 Mar 14;495(7440):223-6
references.pmid: 23467090
references.citation: Proc Natl Acad Sci U S A. 2013 Jul 2;110(27):11091-6
references.pmid: 23776241
references.citation: Trends Immunol. 2009 Jun;30(6):263-70
references.pmid: 19427267
references.citation: J Biol Chem. 2013 Mar 1;288(9):6140-53
references.pmid: 23300080
references.citation: Cancer Res. 1997 May 15;57(10):1835-40
references.pmid: 9157970
references.citation: J Gen Physiol. 2010 Nov;136(5):515-27
references.pmid: 20937692
references.citation: Nature. 2009 Sep 10;461(7261):282-6
references.pmid: 19741708
references.citation: Purinergic Signal. 2009 Jun;5(2):163-73
references.pmid: 19189228
references.citation: J Biol Chem. 2011 Aug 5;286(31):27639-53
references.pmid: 21659516
references.citation: J Cell Sci. 2007 Nov 1;120(Pt 21):3772-83
references.pmid: 17925379
references.citation: Autophagy. 2013 Oct;9(10):1624-5
references.pmid: 23989612
references.citation: Crit Rev Oncol Hematol. 2013 Sep;87(3):306-22
references.pmid: 23541070
references.citation: Cancer Res. 2011 Apr 15;71(8):2892-900
references.pmid: 21292811
references.citation: J Cell Mol Med. 2006 Jul-Sep;10(3):613-34
references.pmid: 16989724
references.citation: Eur J Immunol. 2011 May;41(5):1231-41
references.pmid: 21469131
references.citation: J Physiol. 2012 Dec 15;590(24):6257-66
references.pmid: 23070703
references.citation: FASEB J. 2011 Sep;25(9):3250-61
references.pmid: 21659556
references.citation: Purinergic Signal. 2005 Dec;1(4):311-28
references.pmid: 18404516
references.citation: Cell. 2008 Jun 27;133(7):1149-61
references.pmid: 18585350
references.citation: FEBS Lett. 2014 Apr 17;588(8):1416-22
references.pmid: 24642372
references.citation: Nat Cell Biol. 2007 Aug;9(8):945-53
references.pmid: 17618272
references.citation: J Exp Med. 1998 Sep 7;188(5):979-84
references.pmid: 9730899
references.citation: Mol Biol Cell. 2009 Oct;20(20):4313-23
references.pmid: 19692571
references.citation: Mol Cell. 1999 Feb;3(2):159-67
references.pmid: 10078199
references.citation: Cell Commun Adhes. 2008 May;15(1):119-32
references.pmid: 18649184
references.citation: Biochem Pharmacol. 2011 Nov 1;82(9):1110-25
references.pmid: 21819973
references.citation: Cell Death Differ. 2005 Nov;12(11):1390-7
references.pmid: 15905877
references.citation: Blood. 2010 Nov 4;116(18):3475-84
references.pmid: 20660288