Replication of an integrin targeted conditionally replicating adenovirus on primary ovarian cancer spheroids.
Replication competent viruses hold promise for treatment of advanced cancers resistant to available therapeutic modalities. Although preliminary clinical results have substantiated their efficacy, preclinical development of these novel approaches is limited by assay substrates. The evaluation of candidate agents could be confounded by differences between primary tumor cells and tumor cell lines, as discordance in the levels of surface receptors relevant for viral entry has been reported. Since primary tumor cells are difficult to analyze ex vivo for longitudinal observation of virus replication, we developed three-dimensional aggregates or spheroids of unpassaged and purified ovarian cancer cells as a means for prolonging primary tumor cell viability and as a three-dimensional in vitro model for replicative viral infection. Ovarian cancer cells purified from ascites samples were sustained for 30 days while retaining the infection profile with tropism modified and unmodified adenoviruses (Ads). Cell line and primary cell spheroids were used to quantitate the replication and oncolytic potency of replicative Ads in preclinical testing for human ovarian cancer trials. Therefore, spheroids provide a method to sustain purified unpassaged primary ovarian cancer cells for extended periods and to allow evaluation of replicative viruses in a three-dimensional model.
Lam
John T
JT
Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Bauerschmitz
Gerd J
GJ
Kanerva
Anna
A
Barker
Shannon D
SD
Straughn
J Michael
JM
Wang
Minghui
M
Barnes
Mack N
MN
Blackwell
Jerry L
JL
Siegal
Gene P
GP
Alvarez
Ronald D
RD
Curiel
David T
DT
Hemminki
Akseli
A
D000256:Adenoviridae / Q000502:physiology*; D001201:Ascites; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D016023:Integrins / Q000235:genetics*; D009842:Oligopeptides / Q000235:genetics*; D010051:Ovarian Neoplasms / Q000628:therapy*; D018874:Spheroids, Cellular / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014779:Virus Replication
D016023:Integrins; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid