Employment of liver tissue slice analysis to assay hepatotoxicity linked to replicative and nonreplicative adenoviral agents.
Whereas virotherapy has emerged as a novel and promising approach for neoplastic diseases, appropriate model systems have hampered preclinical evaluation of candidate conditionally replicative adenovirus agents (CRAds) with respect to liver toxicity. This is due to the inability of human viral agents to cross species. We have recently shown the human liver tissue slice model to be a facile means to validate adenoviral replication. On this basis, we sought to determine whether our ex vivo liver tissue slice model could be used to assess CRAd-mediated liver toxicity. We analyzed and compared the toxicity of a conditionally replicative adenovirus (AdDelta24) to that of a replication incompetent adenovirus (Adnull [E1-]) in mouse and human liver tissue slices. To accomplish this, we examined the hepatic apoptosis expression profile by DNA microarray analyses, and compared these results to extracellular release of aminotransferase enzymes, along with direct evidence of apoptosis by caspase-3 immunhistochemical staining and TUNEL assays. Human and mouse liver tissue slices demonstrated a marked increase in extracellular release of aminotransferase enzymes on infection with AdDelta24 compared to Adnull. AdDelta24-mediated liver toxicity was further demonstrated by apoptosis induction, as detected by caspase-3 immunohistochemical staining, TUNEL assay and microarray analysis. In conclusion, concordance of CRAd-mediated apoptosis in both the human and the mouse liver tissue slice models was demonstrated, despite the limited replication ability of CRAds in mouse liver slices. The results of this study, defining the CRAd-mediated apoptosis gene expression profiles in human and mouse liver, may lay a foundation for preclinical liver toxicity analysis of CRAd agents.
Stoff-Khalili
M A
MA
Division of Human Gene Therapy, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294-2172, USA, and Department of Plastic and Reconstructive Surgery, Dreifaltigkeits-Hospital, Wesseling, Germany.
Rivera
A A
AA
Le
L P
LP
Stoff
A
A
Everts
M
M
Contreras
J L
JL
Chen
D
D
Teng
L
L
Rots
M G
MG
Haisma
H J
HJ
Rocconi
R P
RP
Bauerschmitz
G J
GJ
Rein
D T
DT
Yamamoto
M
M
Siegal
G P
GP
Dall
P
P
Michael Mathis
J
J
Curiel
D T
DT
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D001681:Biological Assay; D015536:Down-Regulation; D017353:Gene Deletion; D005822:Genetic Vectors; D006801:Humans; D008099:Liver / Q000166:cytology / Q000821:virology*; D008113:Liver Neoplasms / Q000821:virology*; D051379:Mice; D046228:Microarray Analysis; D008954:Models, Biological; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation; D014779:Virus Replication