Targeting the ERG oncogene with splice-switching oligonucleotides as a novel therapeutic strategy in prostate cancer
Br J Cancer Jun 25. doi: 10.1038/s41416-020-0951-2. Online ahead of print. (2020) -
Authors: Ling Li, Lisa Hobson, Laura Perry, Bethany Clark, Susan Heavey, Aiman Haider, Ashwin Sridhar, Greg Shaw, John Kelly, Alex Freeman, Ian Wilson, Hayley Whitaker, Elmar Nurmemmedov, Sebastian Oltean, Sean Porazinski and Michael Ladomery
BACKGROUND: The ERG oncogene, a member of the ETS family of transcription factor encoding genes, is a genetic driver of
prostate cancer. It is activated through a fusion with the androgen-responsive TMPRSS2 promoter in 50% of cases. There is therefore
significant interest in developing novel therapeutic agents that target ERG. We have taken an antisense approach and designed
morpholino-based oligonucleotides that target ERG by inducing skipping of its constitutive exon 4.
METHODS: We designed antisense morpholino oligonucleotides (splice-switching oligonucleotides, SSOs) that target both the 5′
and 3′ splice sites of ERG’s exon 4. We tested their efficacy in terms of inducing exon 4 skipping in two ERG-positive cell lines, VCaP
prostate cancer cells and MG63 osteosarcoma cells. We measured their effect on cell proliferation, migration and apoptosis. We also
tested their effect on xenograft tumour growth in mice and on ERG protein expression in a human prostate cancer radical
prostatectomy sample ex vivo.
RESULTS: In VCaP cells, both SSOs were effective at inducing exon 4 skipping, which resulted in a reduction of overall ERG protein
levels up to 96 h following a single transfection. SSO-induced ERG reduction decreased cell proliferation, cell migration and
significantly increased apoptosis. We observed a concomitant reduction in protein levels for cyclin D1, c-Myc and the Wnt signalling
pathway member β-catenin as well as a marker of activated Wnt signalling, p-LRP6. We tested the 3′ splice site SSO in MG63
xenografts in mice and observed a reduction in tumour growth. We also demonstrated that the 3′ splice site SSO caused a
reduction in ERG expression in a patient-derived prostate tumour tissue cultured ex vivo.
CONCLUSIONS: We have successfully designed and tested morpholino-based SSOs that cause a marked reduction in ERG
expression, resulting in decreased cell proliferation, a reduced migratory phenotype and increased apoptosis. Our initial tests on
mouse xenografts and a human prostate cancer radical prostatectomy specimen indicate that SSOs can be effective for oncogene
targeting in vivo. As such, this study encourages further in vivo therapeutic studies using SSOs targeting the ERG oncogene.