An adenovirus with enhanced infectivity mediates molecular chemotherapy of ovarian cancer cells and allows imaging of gene expression.
The adenovirus (Ad) is a useful vector for cancer gene therapy due to its unparalleled gene transfer efficiency to dividing and quiescent cells. Primary cancer cells, however, often have highly variable or low levels of the requisite coxsackie-adenovirus receptor (CAR). Also, assessment of gene transfer and vector persistence has been logistically difficult in human clinical trials. We describe here two novel bicistronic adenoviral (Ad) vectors, AdTKSSTR and RGDTKSSTR, which contain the herpes simplex virus thymidine kinase gene (TK) for molecular chemotherapy and bystander effect. In addition, the viruses contain the human somatostatin receptor subtype-2 gene (SSTR2), the expression of which can be noninvasively imaged. We enhanced the infectivity of RGDTKSSTR by genetically incorporating the RGD-4C motif into the HI-loop of the fiber. This allows the virus to circumvent CAR deficiency by binding to alpha(v)beta(3) and alpha(v)beta(5) integrins, which are highly expressed on most ovarian cancers. The expanded tropism of RGDTKSSTR results in increased infectivity of purified primary ovarian cancer cells and allows enhanced gene transfer in the presence of malignant ascites containing anti-Ad antibodies. RGDTKSSTR may be a useful agent for treating ovarian cancer in clinical trials.
Hemminki
A
A
The Gene Therapy Center, Division of Human Gene Therapy, Department of Medicine, University of Alabama at Birmingham, AL 35294, USA. akseli@uab.edu
Belousova
N
N
Zinn
K R
KR
Liu
B
B
Wang
M
M
Chaudhuri
T R
TR
Rogers
B E
BE
Buchsbaum
D J
DJ
Siegal
G P
GP
Barnes
M N
MN
Gomez-Navarro
J
J
Curiel
D T
DT
Alvarez
R D
RD
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology*; D001201:Ascites / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000821:virology; D002470:Cell Survival / Q000187:drug effects; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D004274:DNA, Recombinant / Q000235:genetics; D003952:Diagnostic Imaging / Q000379:methods*; D005260:Female; D015774:Ganciclovir / Q000494:pharmacology; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D005840:Gentian Violet; D006367:HeLa Cells; D006801:Humans; D016254:Mutagenesis, Insertional; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy* / Q000821:virology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017481:Receptors, Somatostatin / Q000235:genetics; D011991:Receptors, Virus / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013937:Thymidine Kinase / Q000235:genetics; D014407:Tumor Cells, Cultured
C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D004274:DNA, Recombinant; D012333:RNA, Messenger; D017481:Receptors, Somatostatin; D011991:Receptors, Virus; C093733:somatostatin receptor 2; D013937:Thymidine Kinase; D005840:Gentian Violet; D015774:Ganciclovir