Optimization of short-term transgene expression by sodium butyrate and ubiquitous chromatin opening elements (UCOEs).
BACKGROUND
Predictable and adequate transgene expression is essential for clinical gene therapy. Several studies have focused on optimization of transgene expression. In this study the effect of sodium butyrate (NaB) and a ubiquitous chromatin opening element (UCOE) on short-term gene expression after adenovirus-mediated gene transfer in fibroblastic interface cells from periprosthetic tissue in loosened orthopedic implants is investigated.


METHODS
Cultures of diploid human interface cells from four patients were infected with an adenovirus type-5 vector that carries the luciferase gene driven by the cytomegalovirus (CMV) promoter as a reporter. In addition, viruses with a UCOE were evaluated. Twenty-four hours after infection NaB was added in concentrations of 0 to 9 mM. Luciferase activity was tested after a further 24 h.


RESULTS
NaB in a concentration of 6 mM caused a 7- to 16-fold increase in reporter gene expression compared to control condition. There was no difference in reporter gene expression when cells were infected with Ad.1.5UCOE-CMV.Luc compared to Ad.CMV.Luc. A combination of NaB and a UCOE had no advantage over NaB alone.


CONCLUSIONS
Addition of NaB results in a marked increase in transgene expression in cultured cells. This would allow the enhancement of the expression of the transgene, without requiring a higher vector dose. Butyrate administration could not be substituted by inclusion of UCOEs in the vector. It remains to be established whether the effective concentrations of butyrate can be obtained in vivo.
de Poorter
Jolanda J
JJ
Leiden University Medical Center, Leiden, The Netherlands.
Lipinski
Kai S
KS
Nelissen
Rob G H H
RG
Huizinga
Tom W J
TW
Hoeben
Rob C
RC
D000256:Adenoviridae / Q000235:genetics; D002087:Butyrates / Q000494:pharmacology*; D002843:Chromatin / Q000737:chemistry / Q000235:genetics*; D003587:Cytomegalovirus / Q000235:genetics; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects* / Q000502:physiology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000494:pharmacology*; D006801:Humans; D008156:Luciferases / Q000235:genetics; D019076:Transgenes / Q000502:physiology*
D002087:Butyrates; D002843:Chromatin; D008156:Luciferases