Prolonged re-expression of the hypermethylated gene EPB41L3 using artificial transcription factors and epigenetic drugs

Christian Huisman*, Monique G. P. van der Wijst, Fahimeh Falahi, Juul Overkamp, Gellert Karsten, Martijn M. Terpstra, Klaas Kok, Ate G. J. van der Zee, Ed Schuuring, G. Bea A. Wisman, Marianne G. Rots

*Bijbehorende auteur voor dit werk

OnderzoeksoutputAcademicpeer review

19 Citaten (Scopus)

Samenvatting

Epigenetic silencing of tumor suppressor genes (TSGs) is considered a significant event in the progression of cancer. For example, EPB41L3, a potential biomarker in cervical cancer, is often silenced by cancer-specific promoter methylation. Artificial transcription factors (ATFs) are unique tools to re-express such silenced TSGs to functional levels; however, the induced effects are considered transient. Here, we aimed to improve the efficiency and sustainability of gene re-expression using engineered zinc fingers fused to VP64 (ZF-ATFs) or DNA methylation modifiers (ZF-Tet2 or ZF-TDG) and/or by co-treatment with epigenetic drugs [5-aza-dC2'-deoxycytidine or Trichostatin A (TSA)]. EPB41L3-ZF effectively bound its methylated endogenous locus, as also confirmed by ChIP-seq. ZF-ATFs reactivated the epigenetically silenced target gene EPB41L3 (˜10-fold) in breast, ovarian, and cervical cancer cell lines. Prolonged high levels of EPB41L3 (˜150-fold) induction could be achieved by short-term co-treatment with epigenetic drugs. Interestingly, for otherwise ineffective ZF-Tet2 or ZF-TDG treatments, TSA facilitated re-expression of EPB41L3 by up to twofold. ATF-mediated re-expression demonstrated a tumor suppressive role for EPB41L3 in cervical cancer cell lines. In conclusion, epigenetic reprogramming provides a novel way to improve sustainability of re-expression of epigenetically silenced promoters.

Originele taal-2English
Pagina's (van-tot)384-396
Aantal pagina's13
TijdschriftEpigenetics
Volume10
Nummer van het tijdschrift5
DOI's
StatusPublished - 4-mei-2015

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