Toxicity associated with repeated administration of first-generation adenovirus vectors does not occur with a helper-dependent vector.
BACKGROUND
Certain gene therapy protocols may require multiple administrations of vectors to achieve therapeutic benefit to the patient. This may be especially relevant for vectors such as adenoviral vectors that do not integrate into the host chromosome. Because immunocompetent animal models used for gene transfer studies develop neutralizing antibodies to adenoviral vectors after a single administration, little is known about how repeat administrations of vectors might affect transgene expression and vector toxicity.


METHODS
We used mice deficient in the membrane spanning region of immunoglobulin (IgM), which do not develop antibodies, to evaluate the effect of repeated intravenous administration of first-generation and helper-dependent adenoviral vectors expressing human alpha 1-antitrypsin (hAAT). The duration and levels of transgene expression were evaluated after repeated administration of vectors. Toxicity was assessed by measuring the level of liver enzymes in the serum and the degrees of hepatocyte hypertrophy and proliferation.


RESULTS
We found that previous administration of first-generation adenoviral vectors can alter the response to subsequent doses. These alterations included an increase in transgene expression early (within 1 and 3 days), followed by a rapid drop in expression by day 7. In addition, previous administrations of first-generation vectors led to an increase in toxicity of subsequent doses, as indicated by a rise in liver enzymes and an increase in hepatocyte proliferation. In contrast to first-generation vectors, use of the helper-dependent adenovirus vector, Ad-STK109, which contained no viral coding regions, did not lead to increased toxicity after multiple administrations.


CONCLUSIONS
We conclude that the response of the host to adenoviral vectors can be altered after repeated administration, compared with the response after the initial vector dose. In addition, these experiments provide further evidence for the relative safety of helper-dependent adenoviral vectors for gene therapy, compared with first-generation vectors.
O'Neal
W K
WK
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA. woneal@email.unc.edu
Zhou
H
H
Morral
N
N
Langston
C
C
Parks
R J
RJ
Graham
F L
FL
Kochanek
S
S
Beaudet
A L
AL
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015870:Gene Expression; D015316:Genetic Therapy / Q000009:adverse effects* / Q000379:methods; D005822:Genetic Vectors; D006378:Helper Viruses / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D007075:Immunoglobulin M / Q000235:genetics; D008099:Liver / Q000201:enzymology / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D012449:Safety; D000515:alpha 1-Antitrypsin / Q000235:genetics
D018398:Homeodomain Proteins; D007075:Immunoglobulin M; D000515:alpha 1-Antitrypsin; C064658:RAG-1 protein