Novel infectivity-enhanced oncolytic adenovirus with a capsid-incorporated dual-imaging moiety for monitoring virotherapy in ovarian cancer.
We sought to develop a cancer-targeted, infectivity-enhanced oncolytic adenovirus that embodies a capsid-labeling fusion for noninvasive dual-modality imaging of ovarian cancer virotherapy. A functional fusion protein composed of fluorescent and nuclear imaging tags was genetically incorporated into the capsid of an infectivity-enhanced conditionally replicative adenovirus. Incorporation of herpes simplex virus thymidine kinase (HSV-tk) and monomeric red fluorescent protein 1 (mRFP1) into the viral capsid and its genomic stability were verified by molecular analyses. Replication and oncolysis were evaluated in ovarian cancer cells. Fusion functionality was confirmed by in vitro gamma camera and fluorescent microscopy imaging. Comparison of tk-mRFP virus to single-modality controls revealed similar replication efficiency and oncolytic potency. Molecular fusion did not abolish enzymatic activity of HSV-tk as the virus effectively phosphorylated thymidine both ex vivo and in vitro. In vitro fluorescence imaging demonstrated a strong correlation between the intensity of fluorescent signal and cytopathic effect in infected ovarian cancer cells, suggesting that fluorescence can be used to monitor viral replication. We have in vitro validated a new infectivity-enhanced oncolytic adenovirus with a dual-imaging modality-labeled capsid, optimized for ovarian cancer virotherapy. The new agent could provide incremental gains toward climbing the barriers for achieving conditionally replicated adenovirus efficacy in human trials.
Kimball
Kristopher J
KJ
Department of Medicine, University of Alabama at Birmingham, USA.
Rivera
Angel A
AA
Zinn
Kurt R
KR
Icyuz
Mert
M
Saini
Vaibhav
V
Li
Jing
J
Zhu
Zeng B
ZB
Siegal
Gene P
GP
Douglas
Joanne T
JT
Curiel
David T
DT
Alvarez
Ronald D
RD
Borovjagin
Anton V
AV
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D002213:Capsid / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000502:physiology; D008856:Microscopy, Fluorescence; D050130:Oncolytic Virotherapy / Q000379:methods*; D010051:Ovarian Neoplasms / Q000628:therapy*; D018139:Simplexvirus / Q000201:enzymology; D013937:Thymidine Kinase / Q000235:genetics / Q000502:physiology; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D013937:Thymidine Kinase; D000097573:Red Fluorescent Protein