PNA-mediated modulation and redirection of Her-2 pre-mRNA splicing: specific skipping of erbB-2 exon 19 coding for the ATP catalytic domain.
The Her-2 receptor coded for by the proto-oncogenic erbB-2 gene is a clinically validated target for treatment of a significant genetic subclass of breast cancers, and Her-2 is also overexpressed or mutated in a range of other cancers. In an approach to exploit antisense mediated splicing interference as a means of manipulating erbB-2 expression in a therapeutically relevant fashion, we have studied the effect on mRNA splicing of a series of peptide nucleic acid (PNA) oligomers targeting specific intron-exon junctions in the erbB-2 pre-mRNA. In particular, we are interested in identifying PNA oligomers that specifically induce skipping of exon 19 as this exon is coding for the ATP catalytic domain of Her-2, and if expressed such truncated version of the Her-2 protein should be functionally inactive in a dominant negative fashion. Therefore, antisense compounds having efficient erbB-2 exon 19 skipping activity could be very interesting in terms of drug discovery. In the present study we identified PNA oligomers having such activity in SK-BR-3 and HeLa cancer cells in culture.
Pankratova
Stanislava
S
Department of Cellular and Molecular Medicine, The Panum Institute, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark.
Nielsen
Birgit N
BN
Shiraishi
Takehiko
T
Nielsen
Peter E
PE
D000255:Adenosine Triphosphate / Q000737:chemistry*; D017398:Alternative Splicing; D020134:Catalytic Domain; D045744:Cell Line, Tumor; D005091:Exons; D005799:Genes, Dominant; D018734:Genes, erbB-2; D006367:HeLa Cells; D006358:Hot Temperature; D006801:Humans; D016376:Oligonucleotides, Antisense; D020135:Peptide Nucleic Acids / Q000378:metabolism*; D012326:RNA Splicing; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
D000255:Adenosine Triphosphate; D016376:Oligonucleotides, Antisense; D020135:Peptide Nucleic Acids; D018719:Erb-b2 Receptor Tyrosine Kinases