Infectivity enhanced adenoviral-mediated mda-7/IL-24 gene therapy for ovarian carcinoma.
OBJECTIVE
Melanoma differentiation associated gene-7 [mda-7/Interleukin (IL)-24] has been identified as a novel anti-cancer agent, which specifically induces apoptosis in cancer cells but not in normal epithelial, endothelial and fibroblast cells. The objective of this study was to evaluate the anti-tumor effect of adenovirus-mediated mda-7/IL-24 (Ad.mda-7) gene therapy in ovarian carcinoma and further improve anti-tumor effect by enhancing infectivity of Ad.mda-7.


METHODS
A panel of human ovarian carcinoma cells, OV-4, HEY, SKOV3, SKOV3.ip1 and control normal human mesothelial cells, were infected by a replication deficient recombinant adenovirus encoding mda-7/IL-24 and control virus Ad.CMV.Luc. After 72 h, apoptosis was evaluated by TUNEL and Hoechst staining and further quantified by fluorescent activated cell sorter (FACS) analysis. Infectivity of Ad.mda-7 was enhanced by retargeting it to CD40 or EGF receptors overexpressed on ovarian cancer cells. Subsequently, enhancement in apoptosis of CD40- or epidermal growth factor receptor (EGFR)-retargeted Ad.mda-7 was evaluated.


RESULTS
Adenoviral-mediated delivery of mda-7 induces apoptosis ranging from 10-23% in human ovarian cancer cells tested with the highest percentage of apoptosis noted in SKOV3 cells. Minimal apoptosis was noted in normal mesothelial cells. CD40- or EGFR-retargeted Ad.mda-7 increased apoptosis by 10-32% when compared to that achieved with untargeted Ad.mda-7.


CONCLUSIONS
Ad.mda-7 exhibits ovarian cancer-specific apoptosis, but does not affect normal human mesothelial cells. Infectivity enhanced CD40- and EGFR-retargeted Ad.mda-7 augments apoptosis induction, thus increasing the therapeutic index and translational potential of Ad.mda-7 gene therapy.
Leath
Charles A
CA
Department of Obstetrics and Gynecology, University of Alabama at Birmingham, 901 19th Street South, Birmingham, AL 35294, USA.
Kataram
Manjula
M
Bhagavatula
Pradeep
P
Gopalkrishnan
Rahul V
RV
Dent
Paul
P
Fisher
Paul B
PB
Pereboev
Alexander
A
Carey
Delicia
D
Lebedeva
Irina V
IV
Haisma
Hidde J
HJ
Alvarez
Ronald D
RD
Curiel
David T
DT
Mahasreshti
Parameshwar J
PJ
D000260:Adenoviruses, Human / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology; D017209:Apoptosis / Q000235:genetics; D019013:CD40 Antigens / Q000378:metabolism; D045744:Cell Line, Tumor; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology / Q000821:virology; D066246:ErbB Receptors / Q000378:metabolism; D005260:Female; D016147:Genes, Tumor Suppressor; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007378:Interleukins / Q000235:genetics* / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000628:therapy* / Q000821:virology; D014779:Virus Replication; D000099180:Interleukin-24
D019013:CD40 Antigens; D066246:ErbB Receptors; D007378:Interleukins; D000099180:Interleukin-24