Retargeted adenoviral cancer gene therapy for tumour cells overexpressing epidermal growth factor receptor or urokinase-type plasminogen activator receptor.
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.
Harvey
T J
TJ
Leeds Institute of Molecular Medicine, St James's University Hospital, Leeds, UK.
Burdon
D
D
Steele
L
L
Ingram
N
N
Hall
G D
GD
Selby
P J
PJ
Vile
R G
RG
Cooper
P A
PA
Shnyder
S D
SD
Chester
J D
JD
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
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
Virology. 2000 Mar 15;268(2):382-90
10704346
Cancer Res. 1999 Jan 15;59(2):325-30
9927041
Lab Anim. 1988 Jul;22(3):195-201
3172698
EMBO J. 1997 May 1;16(9):2294-306
9171344
Curr Cancer Drug Targets. 2009 Feb;9(1):32-71
19200050
Curr Gene Ther. 2004 Sep;4(3):337-46
15384947
Clin Biochem. 2004 Jul;37(7):541-8
15234235
Virology. 2004 Jul 20;325(1):116-28
15231391
Mol Ther. 2003 Sep;8(3):485-94
12946322
Trends Biotechnol. 2007 Jan;25(1):33-9
17084931
Clin Cancer Res. 2008 Sep 15;14(18):5649-55
18794071
J Mol Biol. 2009 May 8;388(3):443-61
19285990
Int J Cancer. 2002 Jun 10;99(5):721-6
12115506
Curr Urol Rep. 2008 Jan;9(1):55-61
18366975
Pharmacol Ther. 2004 Apr;102(1):37-46
15056497
Hum Gene Ther. 2004 Nov;15(11):1034-44
15610604
Anticancer Res. 2003 Mar-Apr;23(2B):1619-23
12820431
J Virol. 2009 Jul;83(13):6416-28
19369326
Clin Cancer Res. 2003 Feb;9(2):619-24
12576427
Cancer Res. 2007 Dec 1;67(23):11309-16
18056457
N Engl J Med. 2008 Oct 23;359(17):1757-65
18946061
Cancer Res. 2002 Aug 1;62(15):4273-81
12154029
Gynecol Oncol. 2002 May;85(2):260-5
11972385
Pharmacogenomics. 2009 Jan;10(1):59-68
19102716
Mol Ther. 2008 Feb;16(2):244-51
18071336
Urology. 2002 Sep;60(3):531-6
12350512
Int J Cancer. 2004 Jan 1;108(1):136-45
14618628
J Virol. 2000 Aug;74(15):6875-84
10888627
Urology. 2003 May;61(5):1053-8
12736046
Science. 1997 Feb 28;275(5304):1320-3
9036860
Mol Ther. 2002 Jun;5(6):695-704
12027553
J Virol. 1998 Nov;72(11):8669-75
9765407
Gene Ther. 2003 Jan;10(2):172-9
12571646
Mol Imaging Biol. 2004 Nov-Dec;6(6):385-94
15564149
Eur J Cancer. 2002 Sep;38(14):1917-26
12204675
World J Urol. 2007 Dec;25(6):573-9
17690890
J Urol. 2002 Jul;168(1):266-72
12050554
Cell. 2008 Feb 8;132(3):397-409
18267072
Cancer Gene Ther. 2005 May;12(5):475-86
15706356
J Virol. 2003 Nov;77(21):11367-77
14557622
Int J Cancer. 1997 Jul 3;72(1):1-22
9212216
Urol Res. 2002 Oct;30(5):301-9
12389118
Trends Biotechnol. 2003 Mar;21(3):117-22
12628368
J Gene Med. 2005 Dec;7(12):1517-25
16170831
Postgrad Med J. 2002 Oct;78(924):584-9
12415079
Urology. 2003 Sep;62(3):559-65
12946777
J Urol. 1993 Aug;150(2 Pt 2):721-5
8326633
BJU Int. 1999 May;83(8):902-8; quiz 908-9
10368227
Cancer Res. 2001 Sep 1;61(17):6592-600
11522659
Cancer Res. 2002 Jul 15;62(14):4157-63
12124355
Gene Ther. 2001 Jul;8(13):969-76
11438831
J Immunol. 2008 Apr 1;180(7):4901-9
18354214
Hum Gene Ther. 2006 Dec;17(12):1187-200
17069538