Promoter- and strain-selective enhancement of gene expression in a mouse skeletal muscle by a polymer excipient Pluronic P85.
Amphiphilic triblock copolymers of ethylene oxide and propylene oxide (Pluronic) significantly enhanced expression of plasmid DNA in the skeletal muscle. In the presence of Pluronic P85 (P85) high levels of expression of a reporter gene (luciferase) were sustained for at least 40 days and the area under the gene expression curve increased by at least 10 times compared to the DNA alone. The effect of Pluronic depended on the strain of the mouse and the type of the promoter used. Thus, P85 enhanced luciferase expression by 17 to 19-fold in immunocompetent C57Bl/6 and Balb/c mice, while no enhancement was observed with athymic Balb/c nu/nu mice. Furthermore, P85 activated the expression of luciferase gene driven by CMV promoter, NFkappaB and p53 response elements. There was much less or no effect on the gene driven by SV40 promoter or AP1 and CRE response elements. Overall, the promoter selectivity suggested that Pluronic induced transcriptional activation of gene expression by activating the p53 and NFkappaB signaling pathways. In addition Pluronic increased the number of DNA copies and thus affected initial stages of gene transfer in a promoter selective manner.
Yang
Zhihui
Z
Center for Drug Delivery and Nanomedicine and Department of Pharmaceutical Sciences, College of Pharmacy, University of Nebraska Medical Center, 985830 Nebraska Medical Center, Omaha, NE 68198-5830, USA.
Zhu
Jian
J
Sriadibhatla
Srikanth
S
Gebhart
Catherine
C
Alakhov
Valery
V
Kabanov
Alexander
A
D000818:Animals; D015153:Blotting, Western; D002626:Chemistry, Pharmaceutical; D002850:Chromatography, Gel; D004247:DNA / Q000008:administration & dosage; D016503:Drug Delivery Systems; D004996:Ethidium; D005079:Excipients; D005260:Female; D005456:Fluorescent Dyes; D015870:Gene Expression / Q000187:drug effects*; D015316:Genetic Therapy / Q000379:methods*; D020672:I-kappa B Proteins / Q000235:genetics; D007202:Indicators and Reagents; D007273:Injections, Intramuscular; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D018482:Muscle, Skeletal / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D023282:Nuclease Protection Assays; D010957:Plasmids / Q000008:administration & dosage; D011060:Poloxalene / Q000737:chemistry / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D005079:Excipients; D005456:Fluorescent Dyes; D020672:I-kappa B Proteins; D007202:Indicators and Reagents; D016328:NF-kappa B; C422648:pluronic block copolymer p85; D011060:Poloxalene; D004247:DNA; D008156:Luciferases; D001616:beta-Galactosidase; D004996:Ethidium