Enhancement of recombinant adeno-associated virus type 2-mediated transgene expression in a lung epithelial cell line by inhibition of the XXXX .

@entity470 @entity12079 @entity669 @entity9504
Recombinant adeno-associated viruses (rAAVs) have attracted considerable interest as gene delivery systems because they show long-term expression in vivo and transduce numerous cell types. Limitations to successful gene transduction from rAAVs have prompted investigations of a variety of treatments to enhance transgene expression from rAAV vectors. @entity12079 -1, an @entity470 ( @entity470 ) @entity669 kinase inhibitor, dramatically enhances rAAV transgene expression. Elegant studies have demonstrated that a single-strand D-sequence-binding protein (ssDBP) is phosphorylated by @entity470 and binds to the D sequence element in the AAV terminal repeat (TR). Binding of the Tyr-phosphorylated ssDBP prevents conversion of single-stranded vector DNA to a double-strand conformation. We observed dramatic increases in transgene expression in lung epithelial cells (IB3) with @entity12079 treatment. Gel shift analysis of ssDBP revealed that its DNA binding characteristics were unchanged after @entity12079 treatment or @entity9504 . @entity12079 stimulated rAAV transgene expression to a greater extent than @entity9504 . Southern hybridizations revealed that the vector DNA remained in the single-strand conformation in @entity12079 -treated cells but double-stranded replicative form monomer DNA was most abundant in adenovirus-infected cells. Northern analyses revealed that @entity12079 treatment enhanced mRNA accumulation more than in adenovirus-infected cultures even though replicative form DNA was undetectable. Analysis of the JNK, ERK, and p38K mitogen-activated protein kinase pathways revealed that @entity12079 treatment stimulated the activity of JNK and p38K. Our data suggest that @entity12079 -induced alteration of stress response pathways results in dramatic enhancement of transcription on linear vector DNA templates in the IB3 cell line. These results expand the downstream targets of the @entity470 in regulating rAAV transduction.
