title: Retargeted adenoviral cancer gene therapy for tumour cells overexpressing epidermal growth factor receptor or urokinase-type plasminogen activator receptor.
abstract: We have assessed the ability of bispecific fusion proteins to improve adenovirus-mediated transfer of therapeutic and marker transgenes. We constructed an expression vector that can be easily modified to synthesize a variety of fusion proteins for retargeting adenoviral gene therapy vectors to cell surface markers, which are differentially expressed between normal and cancer cells. Adenoviral transduction can be improved in a number of tumour cell lines which overexpress EGFR (epidermal growth factor receptor) or uPAR (urokinase-type plasminogen activator receptor), but which have only low levels of endogenous hCAR (human coxsackie B and adenovirus receptor) expression. Up to 40-fold improvement in beta-galactosidase transgene expression was seen using an EGFR retargeting protein, and up to 16-fold using a second fusion protein targeting uPAR. In vitro, our uPAR retargeting fusion protein improved the sensitivity to adenoviral herpes simplex virus thymidine kinase/ganciclovir by an order of magnitude, whereas in vivo, our EGFR retargeting protein is able to significantly delay tumour growth in rodent animal models in a dose-dependent manner. The 'cassette' design of our fusion protein constructs offers a flexible method for the straightforward synthesis of multiple adenoviral retargeting proteins, directed against a variety of tumour-associated antigens, for use in clinical trials.
authors.affiliation: Leeds Institute of Molecular Medicine, St James's University Hospital, Leeds, UK.
authors.forename: T J
authors.initials: TJ
authors.lastname: Harvey
authors.forename: D
authors.initials: D
authors.lastname: Burdon
authors.forename: L
authors.initials: L
authors.lastname: Steele
authors.forename: N
authors.initials: N
authors.lastname: Ingram
authors.forename: G D
authors.initials: GD
authors.lastname: Hall
authors.forename: P J
authors.initials: PJ
authors.lastname: Selby
authors.forename: R G
authors.initials: RG
authors.lastname: Vile
authors.forename: P A
authors.initials: PA
authors.lastname: Cooper
authors.forename: S D
authors.initials: SD
authors.lastname: Shnyder
authors.forename: J D
authors.initials: JD
authors.lastname: Chester
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000998:Antiviral Agents / Q000494:pharmacology; D045744:Cell Line, Tumor; D000090702:Constitutive Androstane Receptor; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D015774:Ganciclovir / Q000494:pharmacology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D009369:Neoplasms / Q000628:therapy*; D015202:Protein Engineering; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D055293:Receptors, Urokinase Plasminogen Activator / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis; D014161:Transduction, Genetic
chemical_list: D000998:Antiviral Agents; D000090702:Constitutive Androstane Receptor; D008565:Membrane Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D055293:Receptors, Urokinase Plasminogen Activator; D011993:Recombinant Fusion Proteins; D066246:ErbB Receptors; D015774:Ganciclovir
references.citation: Virology. 2000 Mar 15;268(2):382-90
references.pmid: 10704346
references.citation: Cancer Res. 1999 Jan 15;59(2):325-30
references.pmid: 9927041
references.citation: Lab Anim. 1988 Jul;22(3):195-201
references.pmid: 3172698
references.citation: EMBO J. 1997 May 1;16(9):2294-306
references.pmid: 9171344
references.citation: Curr Cancer Drug Targets. 2009 Feb;9(1):32-71
references.pmid: 19200050
references.citation: Curr Gene Ther. 2004 Sep;4(3):337-46
references.pmid: 15384947
references.citation: Clin Biochem. 2004 Jul;37(7):541-8
references.pmid: 15234235
references.citation: Virology. 2004 Jul 20;325(1):116-28
references.pmid: 15231391
references.citation: Mol Ther. 2003 Sep;8(3):485-94
references.pmid: 12946322
references.citation: Trends Biotechnol. 2007 Jan;25(1):33-9
references.pmid: 17084931
references.citation: Clin Cancer Res. 2008 Sep 15;14(18):5649-55
references.pmid: 18794071
references.citation: J Mol Biol. 2009 May 8;388(3):443-61
references.pmid: 19285990
references.citation: Int J Cancer. 2002 Jun 10;99(5):721-6
references.pmid: 12115506
references.citation: Curr Urol Rep. 2008 Jan;9(1):55-61
references.pmid: 18366975
references.citation: Pharmacol Ther. 2004 Apr;102(1):37-46
references.pmid: 15056497
references.citation: Hum Gene Ther. 2004 Nov;15(11):1034-44
references.pmid: 15610604
references.citation: Anticancer Res. 2003 Mar-Apr;23(2B):1619-23
references.pmid: 12820431
references.citation: J Virol. 2009 Jul;83(13):6416-28
references.pmid: 19369326
references.citation: Clin Cancer Res. 2003 Feb;9(2):619-24
references.pmid: 12576427
references.citation: Cancer Res. 2007 Dec 1;67(23):11309-16
references.pmid: 18056457
references.citation: N Engl J Med. 2008 Oct 23;359(17):1757-65
references.pmid: 18946061
references.citation: Cancer Res. 2002 Aug 1;62(15):4273-81
references.pmid: 12154029
references.citation: Gynecol Oncol. 2002 May;85(2):260-5
references.pmid: 11972385
references.citation: Pharmacogenomics. 2009 Jan;10(1):59-68
references.pmid: 19102716
references.citation: Mol Ther. 2008 Feb;16(2):244-51
references.pmid: 18071336
references.citation: Urology. 2002 Sep;60(3):531-6
references.pmid: 12350512
references.citation: Int J Cancer. 2004 Jan 1;108(1):136-45
references.pmid: 14618628
references.citation: J Virol. 2000 Aug;74(15):6875-84
references.pmid: 10888627
references.citation: Urology. 2003 May;61(5):1053-8
references.pmid: 12736046
references.citation: Science. 1997 Feb 28;275(5304):1320-3
references.pmid: 9036860
references.citation: Mol Ther. 2002 Jun;5(6):695-704
references.pmid: 12027553
references.citation: J Virol. 1998 Nov;72(11):8669-75
references.pmid: 9765407
references.citation: Gene Ther. 2003 Jan;10(2):172-9
references.pmid: 12571646
references.citation: Mol Imaging Biol. 2004 Nov-Dec;6(6):385-94
references.pmid: 15564149
references.citation: Eur J Cancer. 2002 Sep;38(14):1917-26
references.pmid: 12204675
references.citation: World J Urol. 2007 Dec;25(6):573-9
references.pmid: 17690890
references.citation: J Urol. 2002 Jul;168(1):266-72
references.pmid: 12050554
references.citation: Cell. 2008 Feb 8;132(3):397-409
references.pmid: 18267072
references.citation: Cancer Gene Ther. 2005 May;12(5):475-86
references.pmid: 15706356
references.citation: J Virol. 2003 Nov;77(21):11367-77
references.pmid: 14557622
references.citation: Int J Cancer. 1997 Jul 3;72(1):1-22
references.pmid: 9212216
references.citation: Urol Res. 2002 Oct;30(5):301-9
references.pmid: 12389118
references.citation: Trends Biotechnol. 2003 Mar;21(3):117-22
references.pmid: 12628368
references.citation: J Gene Med. 2005 Dec;7(12):1517-25
references.pmid: 16170831
references.citation: Postgrad Med J. 2002 Oct;78(924):584-9
references.pmid: 12415079
references.citation: Urology. 2003 Sep;62(3):559-65
references.pmid: 12946777
references.citation: J Urol. 1993 Aug;150(2 Pt 2):721-5
references.pmid: 8326633
references.citation: BJU Int. 1999 May;83(8):902-8; quiz 908-9
references.pmid: 10368227
references.citation: Cancer Res. 2001 Sep 1;61(17):6592-600
references.pmid: 11522659
references.citation: Cancer Res. 2002 Jul 15;62(14):4157-63
references.pmid: 12124355
references.citation: Gene Ther. 2001 Jul;8(13):969-76
references.pmid: 11438831
references.citation: J Immunol. 2008 Apr 1;180(7):4901-9
references.pmid: 18354214
references.citation: Hum Gene Ther. 2006 Dec;17(12):1187-200
references.pmid: 17069538