title: The TrkAIII oncoprotein inhibits mitochondrial free radical ROS-induced death of SH-SY5Y neuroblastoma cells by augmenting SOD2 expression and activity at the mitochondria, within the context of a tumour stem cell-like phenotype.
abstract: The developmental and stress-regulated alternative TrkAIII splice variant of the NGF receptor TrkA is expressed by advanced stage human neuroblastomas (NBs), correlates with worse outcome in high TrkA expressing unfavourable tumours and exhibits oncogenic activity in NB models. In the present study, we report that constitutive TrkAIII expression in human SH-SY5Y NB cells inhibits Rotenone, Paraquat and LY83583-induced mitochondrial free radical reactive oxygen species (ROS)-mediated death by stimulating SOD2 expression, increasing mitochondrial SOD2 activity and attenuating mitochondrial free radical ROS production, in association with increased mitochondrial capacity to produce H2O2, within the context of a more tumour stem cell-like phenotype. This effect can be reversed by the specific TrkA tyrosine kinase inhibitor GW441756, by the multi-kinase TrkA inhibitors K252a, CEP-701 and Gö6976, which inhibit SOD2 expression, and by siRNA knockdown of SOD2 expression, which restores the sensitivity of TrkAIII expressing SH-SY5Y cells to Rotenone, Paraquat and LY83583-induced mitochondrial free radical ROS production and ROS-mediated death. The data implicate the novel TrkAIII/SOD2 axis in promoting NB resistance to mitochondrial free radical-mediated death and staminality, and suggest that the combined use of TrkAIII and/or SOD2 inhibitors together with agents that induce mitochondrial free radical ROS-mediated death could provide a therapeutic advantage that may also target the stem cell niche in high TrkA expressing unfavourable NB.
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila, L'Aquila, Italy.
authors.forename: Pierdomenico
authors.initials: P
authors.lastname: Ruggeri
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila, L'Aquila, Italy.
authors.forename: Antonietta R
authors.initials: AR
authors.lastname: Farina
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila, L'Aquila, Italy; Department of Medical-Surgical Science and Biotechnology, University of Rome "La Sapienza", Latina, Italy.
authors.forename: Natalia
authors.initials: N
authors.lastname: Di Ianni
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila, L'Aquila, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Cappabianca
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila, L'Aquila, Italy.
authors.forename: Marzia
authors.initials: M
authors.lastname: Ragone
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila, L'Aquila, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Ianni
authors.affiliation: Department of Experimental Medicine, University of Rome "La Sapienza", Rome, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Gulino
authors.affiliation: Department of Applied Clinical and Biotechnological Sciences, University of L'Aquila, L'Aquila, Italy.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Mackay
mesh_terms: D063646:Carcinogenesis / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D018450:Disease Progression; D019008:Drug Resistance, Neoplasm; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D055785:Gene Knockdown Techniques; D006801:Humans; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000473:pathology*; D009447:Neuroblastoma / Q000473:pathology*; D015513:Oncogene Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D010641:Phenotype; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020917:Receptor, trkA / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018874:Spheroids, Cellular / Q000187:drug effects / Q000473:pathology; D013482:Superoxide Dismutase / Q000172:deficiency / Q000235:genetics*; D000099156:Superoxide Dismutase 2
chemical_list: D016328:NF-kappa B; D015513:Oncogene Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); D020033:Protein Isoforms; D047428:Protein Kinase Inhibitors; D017382:Reactive Oxygen Species; D020917:Receptor, trkA; D013482:Superoxide Dismutase; D000099156:Superoxide Dismutase 2; C051038:Inositol 1,4,5-trisphosphate 3-kinase
references.citation: Nat Protoc. 2010 Jan;5(1):51-66
references.pmid: 20057381
references.citation: J Biol Chem. 2010 Jul 30;285(31):24141-53
references.pmid: 20495008
references.citation: Br J Cancer. 2012 Oct 9;107(8):1409-17
references.pmid: 23047593
references.citation: Mol Cancer Res. 2002 Dec;1(2):137-46
references.pmid: 12496360
references.citation: Cancers (Basel). 2010 May 17;2(2):859-84
references.pmid: 24281098
references.citation: J Neurosci. 1998 Jan 15;18(2):687-97
references.pmid: 9425011
references.citation: J Neurosci. 2007 Oct 17;27(42):11315-26
references.pmid: 17942726
references.citation: J Biol Chem. 1992 Jan 5;267(1):13-6
references.pmid: 1730579
references.citation: Anticancer Agents Med Chem. 2011 Feb;11(2):181-90
references.pmid: 21355846
references.citation: Cancer Lett. 2005 Oct 18;228(1-2):211-9
references.pmid: 15950376
references.citation: PLoS One. 2009;4(1):e4235
references.pmid: 19156211
references.citation: Eur J Med Chem. 2013 Apr;62:187-98
references.pmid: 23353752
references.citation: Mol Cell Neurosci. 2011 Dec;48(4):349-58
references.pmid: 21722732
references.citation: Stem Cells. 2006 Sep;24(9):2110-9
references.pmid: 16728559
references.citation: Cancer Res. 1989 Oct 1;49(19):5239-43
references.pmid: 2766291
references.citation: Eur J Cancer. 1997 Oct;33(12):2068-70
references.pmid: 9516854
references.citation: BMC Biotechnol. 2005 May 10;5:12
references.pmid: 15885144
references.citation: J Neurochem. 1999 Mar;72(3):919-24
references.pmid: 10037462
references.citation: Free Radic Biol Med. 2012 Jun 1-15;52(11-12):2209-22
references.pmid: 22561706
references.citation: Biochem Biophys Res Commun. 2000 Aug 11;274(3):641-8
references.pmid: 10924331
references.citation: Cancer Cell. 2004 Oct;6(4):347-60
references.pmid: 15488758
references.citation: Onco Targets Ther. 2013 Sep 04;6:1207-20
references.pmid: 24043946
references.citation: Cell Growth Differ. 1995 Jun;6(6):727-36
references.pmid: 7669728
references.citation: J Neurosci. 2009 May 27;29(21):7003-14
references.pmid: 19474327
references.citation: J Biol Chem. 1995 Nov 17;270(46):27489-94
references.pmid: 7499206
references.citation: Biochim Biophys Acta. 2001 May 3;1526(2):191-8
references.pmid: 11325541
references.citation: Cancer Res. 2002 Mar 15;62(6):1802-8
references.pmid: 11912158
references.citation: EMBO J. 2002 Apr 2;21(7):1695-703
references.pmid: 11927553
references.citation: Oncogene. 2000 Nov 20;19(49):5690-701
references.pmid: 11114749
references.citation: Oncogene. 2002 May 30;21(24):3917-24
references.pmid: 12032830
references.citation: Chembiochem. 2002 Sep 2;3(9):897-901
references.pmid: 12210991
references.citation: Stem Cells. 2008 Jan;26(1):254-65
references.pmid: 17962704
references.citation: Br J Cancer. 2008 Jul 22;99(2):283-93
references.pmid: 18594523
references.citation: Carcinogenesis. 1997 Jan;18(1):223-8
references.pmid: 9054610
references.citation: Biochem Pharmacol. 2008 Dec 1;76(11):1577-89
references.pmid: 18775681
references.citation: J Biol Chem. 2003 Sep 26;278(39):37832-9
references.pmid: 12853461
references.citation: Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9782-7
references.pmid: 8790408
references.citation: Circulation. 1999 Sep 21;100(12):1330-7
references.pmid: 10491379
references.citation: Oncogene. 2009 Nov 19;28(46):4075-94
references.pmid: 19734938
references.citation: Methods Mol Biol. 2010;648:245-55
references.pmid: 20700717
references.citation: Oncogene. 2012 Mar 8;31(10):1207-16
references.pmid: 21804600
references.citation: J Cell Biol. 1985 Nov;101(5 Pt 1):1799-807
references.pmid: 2997236
references.citation: Free Radic Biol Med. 2008 Apr 15;44(8):1610-6
references.pmid: 18280257
references.citation: Mol Cell Biol. 2000 Aug;20(16):5908-16
references.pmid: 10913174
references.citation: J Biol Chem. 1993 Jul 15;268(20):15150-7
references.pmid: 8325889
references.citation: Free Radic Biol Med. 2009 Aug 15;47(4):344-56
references.pmid: 19477268
references.citation: Cell Biol Int. 2006 Nov;30(11):924-32
references.pmid: 16895761
references.citation: J Biol Chem. 2003 May 9;278(19):16510-9
references.pmid: 12609977
references.citation: Mol Cell Biol. 2009 Sep;29(17):4812-30
references.pmid: 19564412
references.citation: Amino Acids. 2012 Jan;42(1):139-58
references.pmid: 20454814
references.citation: Bioorg Med Chem Lett. 2004 Feb 23;14(4):953-7
references.pmid: 15013000
references.citation: J Biol Chem. 2003 Mar 7;278(10):8516-25
references.pmid: 12496265
references.citation: Proc Natl Acad Sci U S A. 1988 Sep;85(17):6465-7
references.pmid: 3413108
references.citation: Cancer Res. 1979 Apr;39(4):1141-9
references.pmid: 217531
references.citation: Cancer Biol Ther. 2008 Aug;7(8):1271-9
references.pmid: 18497575
references.citation: Toxicology. 2000 Aug 14;149(1):43-50
references.pmid: 10963860
references.citation: Clin Cancer Res. 2003 Jan;9(1):424-32
references.pmid: 12538496
references.citation: J Neuroimmunol. 2007 Feb;183(1-2):151-61
references.pmid: 17241672
references.citation: Blood. 2002 Sep 1;100(5):1828-34
references.pmid: 12176906
references.citation: Biochemistry. 2001 Jul 3;40(26):7890-5
references.pmid: 11425317
references.citation: Biochim Biophys Acta. 2004 Jan 5;1670(1):19-27
references.pmid: 14729138
references.citation: J Biomed Biotechnol. 2010;2010:801726
references.pmid: 20625417
references.citation: Biochemistry. 1998 Feb 10;37(6):1613-22
references.pmid: 9484232
references.citation: Cancer Prev Res (Phila). 2011 May;4(5):638-54
references.pmid: 21543342
references.citation: Oncogene. 2008 Jan 31;27(6):763-74
references.pmid: 17653087
references.citation: Bull Math Biol. 2010 Jul;72(5):1069-91
references.pmid: 19915922
references.citation: Free Radic Biol Med. 2009 Jan 1;46(1):42-50
references.pmid: 18930813
references.citation: Cancer Res. 2007 Nov 1;67(21):10260-7
references.pmid: 17974967
references.citation: Cancer Biol Ther. 2009 Jul;8(13):1300-6
references.pmid: 19458492
references.citation: J Radiat Res. 2012;53(1):58-71
references.pmid: 22302046
references.citation: Curr Biol. 2001 Apr 17;11(8):614-9
references.pmid: 11369207
references.citation: Nat Methods. 2012 Jul;9(7):671-5
references.pmid: 22930834
references.citation: Biomed Res Int. 2013;2013:740187
references.pmid: 23841091
references.citation: Int J Cancer. 1997 Aug 7;72(4):673-9
references.pmid: 9259409
references.citation: Nat Genet. 1995 Dec;11(4):376-81
references.pmid: 7493016
references.citation: J Neurobiol. 1999 May;39(2):323-36
references.pmid: 10235685
references.citation: Nat Protoc. 2008;3(7):1228-39
references.pmid: 18600228